ANKOM TDF Dietary User Manual

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Page 1

Rev E 12/2/14

Operator’s Manual

2052 O’Neil Rd, Macedon NY 14502 Telephone: (315) 986-8090 Fax: (315) 986-8091 www.ankom.com

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Table of Contents

Introduction .................................................................................................................................................................. 5

Warranty ....................................................................................................................................................................... 5

Filter Bags ..................................................................................................................................................................... 5

Contact Information ..................................................................................................................................................... 5

Method Comparison (Manual vs. Automated) ............................................................................................................. 6

Instrument Description ................................................................................................................................................. 7

Service Access Position ............................................................................................................................................... 10

Operating Environment .............................................................................................................................................. 10

Safety Precautions ...................................................................................................................................................... 10

Instrument Installation ............................................................................................................................................... 11

Dietary Fiber Analysis Support Items .......................................................................................................................... 16

Analysis Options using the ANKOM

Blanks .......................................................................................................................................................................... 16

TDF

Dietary Fiber Analyzer ................................................................................... 16

IDF/SDF Analysis ......................................................................................................................................................... 17

TDF Analysis (AOAC 991.43) ....................................................................................................................................... 31

TDF Analysis (AOAC 985.29) ....................................................................................................................................... 45

Protein Determination ................................................................................................................................................ 59

Ash Determination ...................................................................................................................................................... 61

Productivity Enhancement ......................................................................................................................................... 63

Status Screen .............................................................................................................................................................. 67

Fault Handling ............................................................................................................................................................. 71

Diagnostics Mode ....................................................................................................................................................... 73

Periodic Maintenance ................................................................................................................................................. 81

Troubleshooting & Replacement Parts ....................................................................................................................... 81

QC / Calibration Checks .............................................................................................................................................. 83

Appendix A – Reagents (AOAC 991.43, AOAC 985.29) ............................................................................................... 89

Appendix B – Instrument Out-of-use Procedure ........................................................................................................ 90

Appendix C – De-fatting Procedure ............................................................................................................................ 92

Appendix D – Accessories (sold separately) ............................................................................................................... 94

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Operator’s Manual

Please review the entire manual before you begin operating this product.

Introduction

ANKOM Technology designs, manufactures, and markets instruments and support products used by analytical laboratories around the world in the food, feed, bio-energy, agricultural, and environmental industries. ANKOM Technology can provide you with products for determining or monitoring dietary fiber, detergent fiber, crude fiber, fat, digestibility, microbial fermentation (anaerobic or aerobic) and more.

Committed to Total Customer Satisfaction, ANKOM designs every product based on a thorough assessment of customer needs. Congratulations on your purchase of the ANKOM

effectively serve your needs. The ANKOM

reduces technician variation, increasing precision. By carefully following the operating instructions in this manual you will understand the details of sample and filter bag handling as well as instrument control, helping you to achieve the best possible results.

TDF

Dietary Fiber Analyzer was designed to eliminate most of the manual steps required by the method. This

TDF

Dietary Fiber Analyzer. We are confident that this product will

Warranty

ANKOM Technology warrants the ANKOM year after the original date of purchase. This warranty does not include damage to the instrument resulting from neglect or misuse. During the warranty period, should any failure result from defects in workmanship or materials, ANKOM Technology will, at its discretion, repair or replace the instrument free of charge.

TDF

Dietary Fiber Analyzer against any defects in workmanship or material for one

Extended warranties are available upon request.

Filter Bags

Use ANKOM Technology filter bags (part #’s DF-I, DF-S, DF-FT) in your ANKOM be purchased from ANKOM Technology or from your local authorized ANKOM distributor.

TDF

Dietary Fiber Analyzer. Filter bags can

Contact Information

Committed to your total satisfaction, we are always available to help you get the most from your ANKOM products and to receive any comments or suggestions you might have to help us improve.

For any questions or suggestions regarding your instrument, please contact us at:

 Telephone: (315) 986-8090  Fax: (315) 986-8091  Email: service@ankom.com  www.ankom.com

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Operator’s Manual

Prepare for Test

Digest Sample

Isolate IDF Residue

Isolate SDF Residue

Prepare crucibles

5 Heat to 95°C

Transfer to filter

Add 95% EtOH to fitrate

Ash crucibles overnight

6 Add buffer

Filter with vacuum assist

Precipitate for 60 min

Clean with vacuum

7 Add Amylase

Scratch filter bed as needed

Transfer to filter

Soak 1 hr @ room temp

8 Digest at 95°C for 35 min

Filter with vacuum assist

Rinse with 3 solutions

Scrape beaker walls and rinse with H2O to move all residue to bottom

Rinse with 70°C H2O

Scratch filter bed as needed

Prepare DE filter bed

Rinse with 78% EtOH

Dry @ 130°C

Rinse with 95% EtOH

Rinse with 78% EtOH

Cool 1 hr and weigh

Cool to 60°C

Rinse with Acetone

Rinse with 95% EtOH

Weigh samples

Add Protease

Dry IDF residue

Rinse with Acetone

Prepare H2O bath(s)

Digest at 60°C for 30 min

Weigh IDF residue

Dry SDF residue

Place samples in beakers

Add HCl

Weigh SDF residue

Add AMG

Check/adjust pH

Calculate Results

Digest at 60°C for 30 min

Correct for Protein

Correct for Ash

Calculate results using weights and corrections

Prepare for Test

Digest Sample

Isolate IDF Residue

Isolate SDF Residue

5 Heat to 95°C

Transfer to filter

Add 95% EtOH to fitrate

6 Add buffer

Filter with pressure assist

Precipitate for 60 min

7 Add Amylase

Transfer to filter

8 Digest at 95°C for 35 min

Filter with pressure assist

Rinse with 70°C H2O

Rinse with 78% EtOH

Rinse with 95% EtOH

Rinse with 78% EtOH

Weigh filter bags

Cool to 60°C

Rinse with Acetone

Rinse with 95% EtOH

Weigh DE and samples

Add Protease

Dry IDF residue

Rinse with Acetone

Install filter bags

Digest at 60°C for 30 min

Weigh IDF residue

Dry SDF residue

Add DE & Sample to bags

Add HCl

Weigh SDF residue

Add AMG

Check/adjust pH

Calculate Results

Digest at 60°C for 30 min

Correct for Protein

Correct for Ash

Calculate results using weights and corrections

This section is just an overview of how the ANKOM

TDF

Dietary Fiber Analyzer

executes the AOAC 991.43 method. Please see the “IDF/SDF Analysis” and “TDF Analysis” sections of this manual for detailed instructions on how to perform

IDF/SDF and TDF assays with the analyzer.

Method Comparison (Manual vs. Automated)

AOAC 991.43 Method – IDF/SDF assay without automation

Below is the AOAC 991.43 method flow done manually by a technician using glass crucibles and beakers, vacuum to assist with filtering, and water baths to control temperature and mixing. DE refers to Diatomaceous Earth.

AOAC 991.43 Method – IDF/SDF assay with ANKOM

Below is the AOAC 991.43 method flow when using the ANKOM represent tasks done by a technician. Steps highlighted in GREEN (1a-1g, 9, 19, and 30) are no longer needed because of Filter Bag Technology. All other steps (including mixing) are done automatically by the ANKOM

TDF

Dietary Fiber Analyzer automation

TDF

Dietary Fiber Analyzer. Steps highlighted in YELLOW

TDF

Dietary Fiber Analyzer under

computer control. DE refers to Diatomaceous Earth.

pg. 6 Rev E 12/2/14

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Operator’s Manual

Contact ANKOM Technology about an upgrade to Total Integrated Dietary Fiber (AOAC 2009.01 and 2011.25 methods). See the Diagnostics section of this document for instructions on how to enable these methods.

Touch

Screen

Display

Filtrate Cup

(TDF53 - #1 of 6)

Waste Tray

IDF Station (#4 of 6)

SDF Delivery

Nozzle

(#1 of 6)

Clamp Bar A

Clamp Bar B

Control

Buttons

Hook for SDF Bag

(#2 of 6)

IDF Delivery

Nozzle

(#1 of 6)

Control Panel (TDF17)

Front Cover

(Clear)

Clamp Bar D Clamp Bar C

SDF Station (#6 of 6)

Heating Paddle (#1 of 6)

Mixing Pad (#2 of 6)

Service Access

Screw #1

Service Access

Screw #2

Instrument Description

The ANKOM (IDF), Soluble Dietary Fiber (SDF), and Total Dietary Fiber (TDF) residue from food and/or feed samples in accordance with the AOAC 985.29, AOAC 991.43, AACC 32-07.01, and NMKL 129, 2003 methods.

Enabled by Filter Bag Technology, the instrument can run six digestion and six precipitation processes at the same time. The filter bags are designed to capture the appropriate fiber particles while allowing all non-fiber components to pass through. The instrument will use IDF bags and SDF bags when configured to recover IDF and SDF residue. The instrument will use IDF Flow-Thru bags and SDF bags when configured to recover TDF residue.

The automated fiber recovery is achieved by first digesting samples within bags using enzyme treatments. Depending on the filter bag configuration, the instrument then collects the IDF and SDF residue using two separate filters, or collects the TDF residue using one filter. Once captured in the filters the residue is dried, weighed, and corrected for protein and ash content to determine the IDF, SDF, and/or TDF values.

Digestion, precipitation, rinse (H2O and EtOH), and filter operations are performed by the instrument, eliminating manual transfer and filtration steps. An on-board computer precisely controls process temperatures, agitation, and fluid delivery along with digestion, precipitation, and filtration times.

External Components – Front View

TDF

Dietary Fiber Analyzer is designed to efficiently, accurately, and precisely recover Insoluble Dietary Fiber

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Operator’s Manual

Control Panel (TDF17)

95% EtOH Container (TDF42)

HCl Container (TDF38)

Enzyme Containers

(TDF35, TDF36, TDF37)

Nitrogen Supply Port

100 psi (6.9 bar)

Front Cover Arm

Power Switch

Front Cover (TDF56)

78% EtOH Container (TDF41)

Buffer Container (TDF39)

Distilled Water Container (TDF40)

Front Cover Arm

Three Container Holder

Single Container Holder

External Components – Left Side View

External Components – Right Side View

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Operator’s Manual

Check Valve

(1 of 12)

Main Electrical Cabinet

IDF Inlet Valve

Stations 4, 5 & 6

SDF Inlet Valve

Stations 4, 5 & 6

In-Line Heater

Tubing Support Panel

IDF Inlet Valve

Stations 1, 2 & 3

SDF Inlet Valve

Stations 1, 2 & 3

Waste Valves

(1 – 6)

Supply Valves

 Water  Buffer  78% EtOH  95% EtOH

Vacuum Sensor

Supply

Manifold

Clamp Bar Cylinder

(1 of 8)

HCl

Supply Valve

Peristaltic

Pump

Pressure

Sensors

Drain Tube

Supply Valves

 Amylase  Protease  AMG

Torsion Bar

Internal Components – Rear View

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Operator’s Manual

If after running this instrument you plan to leave it unused for 1 week or longer, you must clear out the fluid lines to ensure proper operation. See “Appendix B – Instrument Out-of-use Procedure” for more information.

Hot Surfaces – Do NOT touch the Heating Paddle surfaces during operation. The

surface can exceed 100°C (212°F). Failure to observe this caution may result in

burns.

Hazardous Voltages – Do NOT operate the instrument with the back of the Main

Electrical Cabinet removed. Hazardous voltages are present during operation. Failure

to observe this caution may result in electrical shock or electrocution.

Hazardous Materials – Ethanol is used within this instrument. Follow both local

and federal regulations for vent hood requirements when operating this instrument. Do NOT heat seal or place filter bags in an oven until all acetone has evaporated.

Failure to observe this caution may be hazardous to your health.

WARNING: Attempts to override safety features or to use this instrument in a

manner not specified by ANKOM Technology voids the warranty and may result in serious injury or even death.

This system is designed to meet and/or exceed the applicable standards of CE and CSA.

The Power Switch must be in the OFF position before plugging the instrument Power Cord into the power source.

Please review the entire manual before you begin operating this instrument.

Service Access

Screw #1

Service Access

Screw #2

Main Electrical

Cabinet

Service Access Position

If your instrument must be located in such a way as to restrict access to the back, you can tilt it forward to access the rear of the instrument by removing the two Service Access Screws on the front of the instrument and gently pulling the top of the Main Electrical Cabinet forward and down. The instrument will pivot at its center.

Operating Environment

Your ANKOM

 Ambient Temperature Range: 19°−30°C  Humidity: 20–60% RH  Power: 100V−120V ~ 50/60Hz 15A

TDF

Dietary Fiber Analyzer is designed to operate within the following environments:

220V−240V ~ 50/60Hz 8A

Safety Precautions

pg. 10 Rev E 12/2/14

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Operator’s Manual

Because the instrument is powered by pneumatics, do NOT run it without an adequate Nitrogen Supply. You can run approximately 5 to 10 full assays (IDF/SDF or TDF) with a 304 cu ft tank of nitrogen.

Unpack and place the instrument where it will be used.

The instrument shipping container consists of three separate cardboard pieces: a top, a bottom, and a sleeve that forms the body of the container. Remove the top and the sleeve of the instrument shipping container. Lift the instrument off of the shipping container bottom and place it on a surface that is firm and level in an area near a Nitrogen Supply and a drain or chemical waste container.

Waste fluids are drained from the instrument using gravity. Therefore, position the instrument so that the drain tube can run down from the Waste Tray.

The instrument must not be subjected to excessive shock, vibration, dirt, moisture, oil, or other fluids.

Ethanol is used within this instrument. Follow both local and federal regulations for vent hood requirements when installing this instrument.

Unpack the attachments and accessories.

Within the shipping container is a cardboard box containing attachments and accessories. Open the box and verify that all of the following items are there.

1 – Crescent Wrench (204.9)

1 – 1/8” Allen Wrench (Z303)

1 – T-Handle Torx T20 Driver (Z276)

1 – 3/8” Nut Driver (8416)

2 – Fuses (8305-15A or 8306-8A)

2 – Spray Tips (TDF44)

2 – Container Filters (8202)

1 – Buffer Filter (8203)

1 – Waste Tray

1 – Drain Tube (665)

1 – Drain Tube Clamp (8287)

2 – Drain Tube Elbow Connectors (8288)

Instrument Installation

Site Requirements

To install and operate the ANKOM

 Adequate power (see “Operating Environment” section of this manual)  Drain or waste container to capture waste fluids  100 psi (6.9 bar) Nitrogen Supply and connection tubing

Instrument Installation Procedure

To install the ANKOM

TDF

Dietary Fiber Analyzer, follow the steps below.

TDF

Dietary Fiber Analyzer you will need the following:

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Operator’s Manual

1 – Pinch Valve Tubing Set (TDF71)

1 – Pump Tube Set (TDF68)

1 - Line Flush Tubing assy (TDF70)

1 – Amylase Container assy (TDF35)

1 – Protease Container assy (TDF36)

1 – AMG Container assy (TDF37) 1 – HCl Container assy (TDF38)

1 – Mes-Tris Buffer Container assy (TDF39)

1 – Distilled Water Container assy (TDF40)

1 – 78% EtOH Container assy (TDF41)

1 – 95% EtOH Container assy (TDF42)

6 – Filtrate Cups (TDF53)

1 – Control Panel assy (TDF17)

1 – Bag Weigh Holder assy (TDF52)

1 – Drying Rack assy (TDF50)

2 – Desiccant Pouches (X45)

1 – Air Tube (8216)

1 – CD with Calculation Templates

1 – three container holder

1 – single container holder

1 – Acetic Acid Container assy (TDF59) &

Fluid Tube Label

(ONLY with AOAC 2009.01/2011.25 methods)

pg. 12 Rev E 12/2/14

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Operator’s Manual

1 – Amylase/AMG Container assy (TDF57)

(ONLY with AOAC 2009.01/2011.25 methods)

1 – Trizma Container assy (TDF58)

(ONLY with AOAC 2009.01/2011.25 methods)

1 – Maleate Buffer Container assy (TDF39) &

Fluid Tube Label

(ONLY with AOAC 2009.01/2011.25 methods)

1 - Enzyme Pinch Bar (8167)

1 - Spray Tip Cleaning Tool (8130)

Place the Waste Tray under the front of the instrument.

With the Drain Tube connector pointing away from you, slide the tray under the front of the instrument until it cannot go back any farther. (When the tray is in place, the side ridges will contact the front face of the steel frame.)

Place the six Filtrate Cups on the Waste Tray if you want to capture the SDF filtrate.

Connect the Drain Tube to the Waste Tray.

Connect and secure the Drain Tube to the Waste Tray using the supplied Drain Tube Clamp. Run it to either a drain or a waste container, avoiding twists in the tube. If necessary cut the Drain Tube and use the Elbow Connectors to meet the needs of your location.

Connect the Chemical Container Holders to the feet of the instrument.

Remove the nuts from the outside of the left and right feet of the instrument. Slide the single container holder onto the bolts on the left foot and the three container holder onto the bolts on the right foot. Secure the holders using the nuts.

Bolts to attach the

three container holder

Bolts to attach the

single container holder

Elbow Connector

Drain Tube Clamp

Rev E 12/2/14 pg. 13

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Operator’s Manual

Connect the Control Panel to the instrument.

Remove the nuts from the right side of the instrument. Connect the 25 pin connector from the instrument to the Control Panel. Slide the Control Panel onto the bolts and secure it in place using the nuts.

Place the Chemical Containers in their holders.

If you are going to run an IDF/SDF or TDF analysis immediately after installing the instrument, fill the Chemical Containers above the Minimum Level lines and fill the Enzyme Containers with at least 10 ml of solution before connecting the containers to the instrument. See Appendix A for the chemical preparations.

Do NOT fill the Chemical and Enzyme Containers if the instrument is going to sit unused after installation.

Place the HCl Container in the holder on the left side of the instrument. In the holder on the right side of the instrument, place the Distilled Water Container in the front section, the Buffer Container in the middle section, and the 78% EtOH Container in the back section. The 95% EtOH Container sits on the bench behind the container holder.

To ensure proper fluid delivery, you must use the Chemical Containers provided with the instrument, and place them in their specified locations.

Connect the Chemical Containers to the corresponding tubes.

The Chemical Containers and associated tubes are all labeled and color-coded for easy connection. The large containers on the right side of the instrument also have push-on connectors. Make sure that you hear a click sound when you connect the tubes to the containers.

On the left side of the instrument is a black tube with an HCl label that has a white barbed fitting on its end. Push the white barbed fitting into the black tube connected to the HCl container.

Push-on Connector

Bolts for

Control Panel

25 pin connector

Barbed Fitting

Black tube connected to HCl container

pg. 14 Rev E 12/2/14

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Operator’s Manual

10.

Connect the Enzyme Containers to the corresponding ports.

The Enzyme Containers connect into labeled, color-coded ports on the left side of the instrument using push-on connectors. Make sure that you hear a click sound when you connect the enzyme containers to the ports.

11.

Connect the Nitrogen Supply to the Nitrogen Supply Port.

The Nitrogen Supply Port has a Push-to-Connect fitting that accepts

¼” OD tubing. Acceptable tubing materials include Nylon,

Polyurethane, Polyolefin, and Teflon. The tubing material must be suitable for the specified pressure.

12.

Set up the Peristaltic Pump.

Unscrew the six acorn nuts on the back of the instrument and remove the clear back panel to access the pump area. You might have to tilt the instrument forward into Service Access Position to access the top two nuts located behind the black torsion bar. Clip and remove the cable tie from around the pump. Latch the two pump doors so they are snapped closed.

13.

Reinstall the clear back panel and tighten the acorn nuts.

Push-on Connector

Nitrogen Supply Port

100 psi (6.9 bar)

Acorn Nuts (the top two are behind the black torsion bar)

Cable Tie

Pump Doors

Rev E 12/2/14 pg. 15

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Operator’s Manual

Item

ANKOM Part #

Electronic Balance with four-place readout

Electronic Balance Software

TBS

Heat Sealer for sealing the filter bags

1915 (120V), 1920 (220V)

IDF Filter Bags

DF-I

SDF Filter Bags (used for SDF and TDF procedures)

DF-S

IDF Flow-Thru Bags (TDF procedure only)

DF-FT

Solvent Resistant Marker

F08

Rinse Stand

TDF51

Diatomaceous Earth (DE)

XTC

Drying Oven

Ashing Oven

Protein Determination Equipment (Kjeldahl is recommended)

Analysis Name

Analysis Description

Upper Bag

Lower Bag

IDF/SDF (991.43)

This analysis is used to determine IDF and SDF residue separately using two different filter bags. TDF is calculated by adding the IDF and SDF residue weights corrected for protein and ash content.

IDF

SDF

TDF (991.43 & 985.29)

This analysis is used to determine TDF residue in one filter. In this analysis, the TDF value consists of the weighed residue corrected for protein and ash.

IDF Flow-Thru

(no filter)

SDF

Contact ANKOM about an upgrade to Total Integrated Dietary Fiber (AOAC

2009.01 and 2011.25 methods). See the Diagnostics section of this document for instructions on how to enable these methods.

Dietary Fiber Analysis Support Items

The following support items needed to perform the fiber analysis may be purchased separately:

Analysis Options using the ANKOM

The ANKOM following describes these analyses along with their specific filter bag configurations.

A computer controlled Diagnostics mode is provided through the Control Panel for maintenance and troubleshooting purposes. To maximize productivity, this instrument also allows a user to start new procedures (IDF/SDF and TDF) before the previous

ones have completed. See the “Productivity Enhancement” section of this manual for details.

TDF

Dietary Fiber Analyzer can be configured to recover IDF and SDF residue, or to recover TDF residue. The

Dietary Fiber Analyzer

Blanks

BLANK values are used in the IDF, SDF, and TDF calculations. The following BLANK values (based on extensive research done in the ANKOM lab) are the default values loaded in the IDF_SDF and TDF spreadsheets in the MS-Excel workbook provided with the instrument.

IDF (AOAC 991.43 method): -0.0072 TDF (AOAC 991.43 method): -0.0047 SDF (AOAC 991.43 method): -0.0030 TDF (AOAC 985.29 method): -0.0028

You do not need to run BLANKS with every set of samples. However, if you want to determine your own BLANK values, especially when you change chemical lots, ANKOM recommends that the BLANK value in the IDF, SDF, and TDF

calculations be the average of at least 18 BLANK values.

If you determine your own BLANK values, you can overwrite the values currently in the MS-Excel spreadsheets.

TDF

pg. 16 Rev E 12/2/14

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Operator’s Manual

To enhance the productivity of your instrument, you can begin the IDF process of a new run while the SDF process of a previous run is finishing. See the “Productivity Enhancement” section of this manual for more details.

Using a Bag Weigh Holder is critical to eliminate the effects of static electricity during the weighing process.

Because different balances have different sensitivities, the Bag Weigh Holder should be placed in the center of the balance for best results.

IDF Bag (shorter bag)

SDF Bag (longer bag)

IDF/SDF Analysis

An IDF/SDF analysis measures the amount of IDF and SDF within a given sample. This requires the use of ANKOM DF-I and DF-S filter bags. When starting a new run, the ANKOM total of twelve bags).

To perform an IDF/SDF analysis, follow the steps below.

TDF

Dietary Fiber Analyzer must have bags installed at all stations (a

1. Prepare chemicals and enzymes.

When using the ANKOM methods use the chemicals and enzymes referred to therein. See Appendix A of this manual for the list of chemicals and enzymes and the instructions for how to prepare them for use in this instrument.

TDF

Dietary Fiber Analyzer for the AOAC 991.43, AACC 32-07.01, and NMKL 129,2003

2. De-fat (see "Appendix C – De-fatting Procedure" in this manual) and de-sugar your samples as

needed according to the AOAC 991.43, AACC 32-07.01, or NMKL 129,2003 methods.

3. Label the filter bags using a Solvent Resistant Marker.

4. Prepare for data collection.

You will need a place to store the data collected during this analysis. For your convenience a CD titled “Calculation Templates” was included with the instrument. This CD has data spreadsheets that can be used for this analysis. Please read the “Instructions” tab in the MS-Excel file for information about the spreadsheets.

5. Weigh filter bags.

Roll or fold each bag and place it in a tared Bag Weigh Holder. Place the Bag Weigh Holder in the center of a balance and record the weight.

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Operator’s Manual

Although the instrument works in accordance with AACC and NMKL methods, the Touch Screen Display only refers to the AOAC methods.

←

Diagnostics

Touch Screen Display version: A.AA SPLat controller software version: vX.XX dd-mm-yy

Pump Tube

Test

Expert Mode

(OFF)

Faults

Service

Mode

Clear IDF /

SDF lines

Digestion

Times

Heater Test

Temperatures

Pressures

Line Charge

Motor Test

Valve Test

Select a Function

AOAC 991.43

IDF/SDF

AOAC 985.29

TDF

Diagnostics

AOAC 991.43

TDF

6. Weigh Diatomaceous Earth (DE).

DE is used during fiber analysis to enhance the flocculation and filtration of the SDF fraction. Place ca 1 g of DE in each of six tared and numbered tins and record the weights.

7. Weigh Samples.

Place 0.50.05 g of sample in each of six tared and numbered tins and record the weights.

8. Turn the instrument power on.

When you turn the power on, the instrument will run through an initialization process and the Control Panel will turn on.

9. On the Touch Screen Display, select the instrument function you would like to perform.

The Control Panel on the ANKOM you will press identified buttons on the Touch Screen Display and buttons below the screen.

When your instrument is initialized and ready to operate, the following screen will be displayed.

TDF

Dietary Fiber Analyzer uses Touch Screen technology. To operate the instrument

9.1 If this is the first time the instrument is being operated after being installed, or after sitting unused for a period of

time (see Appendix B), or after tubing was replaced, the lines must be charged to avoid a Vacuum Sensor Fault. To charge the lines follow the steps below.

9.1.1 Press the Diagnostics button on the “Select a Function” screen. The following screen will be displayed.

9.1.2 Press the Line Charge button. The following screen will be displayed.

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Operator’s Manual

SDF Bags (and clamp bar D) installed?

AOAC 991.43 IDF/SDF

←

Select a Function

AOAC 991.43

IDF/SDF

AOAC 985.29

TDF

Diagnostics

AOAC 991.43

TDF

←

Line Charge

Amylase

Protease

AMG

HCl

EtOH78

EtOH95

Water

Buffer

ALL

9.1.3 Press the ALL button to charge all of the lines. Each button will change color as the associated line is being

charged. All lines are charged when all of the buttons return to their original green color.

9.1.4 Press the back button twice. The following “Select a Function” screen will be displayed again.

9.2 Press the AOAC 991.43 IDF/SDF button. The following screen will be displayed.

Rev E 12/2/14 pg. 19

Notice that the function you selected is now displayed in the top right corner of the screen and a back button is displayed in the top left corner of the screen.

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Operator’s Manual

As part of normal operation, solution from the IDF bag will flow into the SDF bag. Therefore, when installing the SDF filter bags it is very important to position them high enough vertically so that at least 20 mm (0.75 inches) of the filter part of the IDF bag can fit inside the top of the SDF bag.

Clamp Bar C

Clamp Bar D

Clamp Bar B

Clamp Bar A

10. Install SDF filter bags on the ANKOM

10.1 Remove Clamp Bars A, B, C, and D by lifting them off of the locator rods.

10.2 Gently pull the black SDF Delivery Nozzle out toward you.

TDF

Dietary Fiber Analyzer.

10.3 Place a labeled and weighed SDF bag up underneath the SDF Delivery Nozzle so that the Delivery Nozzle is

inside the top part of the bag. Pull the bag up so that the top of the bag is about 35 mm (1.375 inches) above the top of Clamp Bar C and return the Delivery Nozzle to its original position. This will hold the back of the bag in place.

pg. 20 Rev E 12/2/14

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Operator’s Manual

When adding DE to the filter bags it is very important to keep it below the tip of the Delivery Nozzle so that the DE material can be properly rinsed.

Diatomaceous Earth

added in SDF Bags?

AOAC 991.43 IDF/SDF

←

Centering Lines

10.4 With the bag being held by the Delivery Nozzle, center it horizontally between the lines on the back part of

Clamp Bar C.

10.5 Repeat steps 10.2 – 10.4 for all six stations.

10.6 Re-install Clamp Bar D by setting it on the locator rods. Make sure the letter is on the top of the bar and the

rubber material is facing in toward the instrument.

10.7 Flatten the bags to remove any wrinkles.

10.8 With fingers away from the clamp bars, press the check mark () button on the Touch Screen Display to pinch

the bags just above the filter. The following screen will be displayed.

11. Add DE to the SDF filter bags on the ANKOM

11.1 Open the top of the SDF bag and add a weigh tin of DE to the bag by folding the tin and then dipping it down into

the bag below the tip of the Delivery Nozzle.

TDF

Dietary Fiber Analyzer.

11.2 Rinse the tin with no more than 3 ml of Distilled Water to ensure complete transfer.

11.3 Repeat steps 11.1 – 11.2 for all six stations.

Rev E 12/2/14 pg. 21

Page 22

Operator’s Manual

After you confirm that the DE is added, the Clamp Bar D button on the Control Panel is disabled until the SDF process completes.

For proper mixing during the IDF process the IDF bags must be horizontally centered over the Heating Paddles and between the Centering Lines on the back part of Clamp Bar A.

IDF Bags (and clamp bar B) installed?

AOAC 991.43 IDF/SDF

←

Centering Lines

Heating Paddle

11.4 Press the check mark () button on the Touch Screen Display. The following screen will be displayed.

12. Install IDF filter bags on the ANKOM

12.1 Gently pull the black IDF Delivery Nozzle out toward you.

12.2 Place a labeled and weighed IDF bag up underneath the Delivery Nozzle so that the Delivery Nozzle is inside the

top part of the bag. Pull the bag up so that the top of the filter part of the IDF bag is just below the bottom of Clamp Bar B and return the Delivery Nozzle to its original position. This will hold the back of the bag in place.

12.3 With the bag held by the Delivery Nozzle, center it horizontally between the Centering Lines on the back part of

Clamp Bar A.

TDF

Dietary Fiber Analyzer.

pg. 22 Rev E 12/2/14

Page 23

Operator’s Manual

When adding sample to the filter bags it is very important to keep it below the tip of the Delivery Nozzle so that it can be properly rinsed.

Samples in IDF Bags?

AOAC 991.43 IDF/SDF

←

Clamp Bar B

Top part of the filter is positioned just below the bottom of Clamp Bar B

20 mm (0.75 inches) of the IDF filter inside the top of the SDF bag

Heating Paddle

12.4 Place at least 20 mm (0.75 inches) of the filter section of the IDF bag inside the top of the SDF bag to allow for

the flow of liquid into the SDF bag after the IDF process is complete.

12.5 Repeat steps 12.1 – 12.4 for all six stations.

12.6 Re-install Clamp Bar B by setting it on the locator rods. Make sure the letter is on the top of the bar and the

rubber material is facing in toward the instrument.

12.7 Flatten the IDF bags to remove any wrinkles.

12.8 With fingers away from the clamp bars, press the check mark () button on the Touch Screen Display to pinch

the IDF bags just above the filter. The mixing pads will make contact with the bags when Clamp Bar B pinches the IDF bags. The following screen will be displayed.

13. Add samples to the IDF bags.

13.1 Open the top of the IDF bag and transfer the sample from a weigh tin into the IDF bag by folding the tin and then

dipping it down into the bag below the tip of the Delivery Nozzle.

Rev E 12/2/14 pg. 23

Page 24

Operator’s Manual

Hooking the bags in place will put a physical hole in the front of each SDF bag.

Front of SDF Bags hooked to clamp bar C?

AOAC 991.43 IDF/SDF

←

All remaining clamp bars installed?

AOAC 991.43 IDF/SDF

←

Hook on front part of Clamp Bar C

13.2 Rinse the tin with no more than 3 ml of Distilled Water to ensure complete transfer.

13.3 Repeat steps 13.1 – 13.2 for all IDF stations.

13.4 Re-install Clamp Bar A by setting it on the locator rods. Make sure the letter is on the top of the bar and the

rubber material is facing in toward the instrument.

13.5 Press the check mark () button on the Touch Screen Display. The following screen will be displayed.

13.6 Make sure that all clamp bars are installed with the letter on the top of the bar and the rubber material facing in

toward the instrument. Press the check mark () button on the Touch Screen Display. The following screen will be displayed.

14. Hook the front of each SDF bag in place.

14.1 Secure the SDF filter bags in place with the hooks located on the front part of Clamp Bar C.

14.2 Press the check mark () button on the Touch Screen Display. If you ran a method other than 991.43 prior to this

run, the following screen will be displayed.

pg. 24 Rev E 12/2/14

Page 25

Operator’s Manual

Do NOT leave the enzyme ports on the instrument open to the air or the enzymes in the valves may dry up and plug the ports.

Nitrogen supply ON?

AOAC 991.43 IDF/SDF

←

Fluid levels correct?

AOAC 991.43 IDF/SDF

←

Amylase -> Amylase AMG -> AMG HCL -> HCL Buffer -> Mes Tris Buffer

AOAC 991.43 IDF/SDF

←

Check supply containers

15. Verify that the fluid containers have the correct fluids for this procedure.

Verify that the fluid supply containers are configured according to the screen above and press the check mark () button. The following screen will be displayed.

16. Fill fluid containers.

To ensure that you have enough fluids to run a complete IDF/SDF procedure, you must begin with fluid levels above the Minimum Level lines on the chemical containers and at least 10 ml of each enzyme. Add fluids and enzymes as necessary.

With all fluid containers filled to the proper levels, press the check mark () button on the Touch Screen Display. The following screen will be displayed.

Rev E 12/2/14 pg. 25

Page 26

Operator’s Manual

The initial filter times shown when you first run the instrument are based on factory experience. During filtration the computer allows you to bypass the filter time if you notice the filtering is complete. The computer also allows you to add time during filtering if needed (see the “Status Screen” section of this manual for more detail).

Check pH manually?

AOAC 991.43 IDF/SDF

←

Yes

Filter minutes OK?

AOAC 991.43 IDF/SDF

←

After release

After water rinses

After EtOH rinses

After release

After EtOH rinses

5.0

1.0

10.0

3.0

IDF

SDF

(Press button with number to change)

17. Connect the Nitrogen Supply to the instrument and turn on.

Make sure the Nitrogen supply in your lab is connected to the instrument and turned on. The pressure gauges on the instrument should show 50-55 psi on the left and about 4 psi on the right.

Press the check mark () button on the Touch Screen Display. The following screen will be displayed.

18. Set filter times (in minutes).

Because different samples take different amounts of time to filter, the above screen allows you to set your filter times. To change any of the times shown on the screen, press the specific gray button. A number pad will be displayed that will allow you to enter the time that you want. The times you enter will remain until you change them again.

When all of the times shown on the screen are what you want, press the check mark () button on the Touch Screen Display. The following screen will be displayed.

19. Set the Manual pH Check.

If you plan to check the pH after the required HCl is added (during the IDF process), press the Yes button on the screen above. Otherwise, press the No button. The following screen will be displayed.

pg. 26 Rev E 12/2/14

Page 27

Operator’s Manual

Testing pump tubing.

Please wait..........

Initializing lines…

Amylase

EtOH78

Protease

EtOH95

AMG

Water

HCL

Buffer

Vacuum sensor:

AAAA

psi

Press START to begin

AOAC 991.43 IDF/SDF

←

START

20. Start the instrument.

The instrument is now completely set up and ready to automatically run the IDF/SDF procedure. Press the START button to begin. At the beginning of each new run, the instrument automatically runs a tube integrity test.

For the first run after a power-up cycle, or for the first run after the instrument has been idle for twelve hours, the following screen will be displayed.

If the fluid lines are already charged properly, or when the line charge is complete, the following screen will be displayed.

When the tube integrity check is complete, the instrument will automatically execute the IDF/SDF procedure stopping only for faults, aborts, and manual pH measurement (if enabled). The Status screen will show actions and faults as they occur during the automatic operation.

Rev E 12/2/14 pg. 27

Page 28

Operator’s Manual

When measuring pH, use a probe that can be easily rinsed (with Distilled Water) to avoid loss of sample. If you add acid or base to adjust the pH, you must mix the solution in order to get an accurate pH reading. To manually mix the solution, press the outside of the bag with your fingers just above the clamp bar multiple times.

22.1 Remove the IDF bags from the instrument.

22.2 Place the bags on the Rinse Stand by sliding the back part of each bag under the

pinch mechanism. Keep the top of the bag open.

22.3 Using a wash bottle, squirt acetone into each bag, making sure that all residue on

the clear polypropylene surfaces is rinsed down into the filter.

22.4 Repeat step 22.3 so that each bag gets rinsed a total of two times.

22.5 Allow acetone to evaporate from the bags.

Hazardous Materials – Do NOT heat seal or place bags in an oven until all

acetone has evaporated.

Status

AOAC 991.43 IDF/SDF

Overall Progress

Action in progress

IDF

Time remaining:

Amylase/AMG Phase

XX:XX:XX

Time remaining: XX:XX:XX

SDF

Time remaining:

XX:XX:XX

pH required?

Display

Temps/Pressures YES

IDF

ABORT

PAUSE

SDF

ABORT

Check pH

then press continue

xx:xx:xx

Continue

21. Manually measure pH.

One of the questions you are asked before starting the automated procedure is: “Check pH manually?” If you answered “Yes” to this question, the instrument will stop after adding the required HCl, open Clamp Bar A, display the screen

below, and make a sound to remind you to manually measure pH and adjust if necessary.

When you have completed the pH measurement process, press the Continue button on the Status screen. You will see “COMPLETE” next to “pH required?” in the upper left corner of the screen.

22. Rinse the IDF filter bags with acetone.

After the instrument has completed an IDF process the IDF residue that has been collected in the IDF bag must be manually rinsed twice with acetone. It is recommended that you use an ANKOM TDF51 Rinse Stand (sold separately) for rinsing filter bags with acetone.

To rinse the IDF filter bags with acetone using the ANKOM TDF51 Rinse Stand, follow the steps below.

pg. 28 Rev E 12/2/14

Page 29

Operator’s Manual

22.6 With your Heat Sealer set between 3 and 4 (settings may vary depending on

the heat sealer and the power source), press the Heat Sealer arm down for 3 to 4 seconds to seal each bag just above the filter. This keeps all residue contained to the filter area while handling the bags.

22.7 Place each bag in the Drying Rack.

Hazardous Materials – Do NOT heat seal or place bags in an oven until all

acetone has evaporated.

Sealing each bag as close to the filter as possible allows you to trim the maximum amount of polypropylene from the bag before determining the protein content.

23. Rinse the SDF filter bags with acetone.

After the instrument has completed an SDF process the SDF residue that has been collected in the SDF bag must be manually rinsed with acetone. It is recommended that you use an ANKOM TDF51 Rinse Stand for rinsing filter bags with acetone.

To rinse the SDF filter bags with acetone using the ANKOM TDF51 Rinse Stand, follow the steps below.

23.1 Remove the SDF bags from the instrument.

23.2 Place the bags on the Rinse Stand by sliding the back part of each bag under the pinch mechanism. Keep the top

of the bag open.

23.3 Using a wash bottle, squirt acetone into each bag, making sure that all residue on the clear polypropylene surfaces

is rinsed down into the filter.

23.4 Repeat step 23.3 so that each bag gets rinsed a total of two times.

23.5 Allow acetone to evaporate from the bags.

23.6 Seal each bag just above the filter to keep all residue contained to the filter area while handling the bags.

23.7 Place each bag in the Drying Rack.

24. Dry the IDF and SDF filter bags.

24.1 Make sure your oven is 105°C at the location where the bags will be placed.

24.2 Place the Drying Rack with the filter bags in the oven and dry to constant weight (about 90 minutes).

24.3 When dry, remove all IDF and SDF filter bags from the oven and place them in separate desiccant pouches.

25. Weigh the IDF residue.

25.1 Remove only one IDF filter bag from the desiccant pouch at a time.

25.2 Roll or fold the bag, place it in a tared Bag Weigh Holder (BWH), and place the BWH in the center of a balance.

25.3 Record the weight of the filter bag.

25.4 Repeat steps 25.1 – 25.3 for each IDF filter bag used in the fiber analysis.

26. Weigh the SDF residue.

26.1 Remove only one SDF filter bag from the desiccant pouch at a time.

26.2 Roll or fold the bag, place it in a tared Bag Weigh Holder (BWH), and place the BWH in the center of a balance.

26.3 Record the weight of the filter bag.

26.4 Repeat steps 26.1 – 26.3 for each SDF filter bag used in the fiber analysis.

Rev E 12/2/14 pg. 29

Page 30

Operator’s Manual

% IDF = [(R1 + R2)/2] - P - A - B

(M1 + M2)/2

[(( fF1 - fS1) + ( fF2 - fS2))/2] - P - A - B

(M1 + M2)/2

Where:

M1, M2

Original wt of duplicate samples adjusted for pre-treatment fat and sugar losses (g)

R1, R2

Residue for duplicate samples (g)

Final Filter Bag with residue (g)

Initial Filter Bag (g)

P =

Protein of residue and bag (g)

Ash of residue and bag (g)

Blank (g)

= [(BR1 + BR2)/2] - PB - AB

= [((f

BF1

- f

BS1

) + (f

BF2

- f

BS2

))/2] - PB - AB

BR1, BR2

Residue for duplicate blanks (g)

fBF

Final Blank Filter Bag (g)

fBS

Initial Blank Filter Bag (g)

Protein of Blank Filter Bag (g)

Ash of Blank Filter Bag (g)

% SDF

[(R1 + R2)/2] - P - A - B

(M1 + M2)/2

[(( fF1 - fS1 - D1) + ( fF2 - fS2 - D2))/2] - P1 - (A2 - D2) - B

(M1 + M2)/2

Where:

M1, M2

Original wt of duplicate samples adjusted for pre-treatment fat and sugar losses (g)

R1, R2

Residue for duplicate samples (g)

Final Filter Bag with residue (g)

Initial Filter Bag (g)

D =

Original wt of Diatomaceous Earth (g)

P =

Protein of residue and bag (g)

Ash of residue and bag (g)

Blank (g)

= [(BR1 + BR2)/2] - PB - (AB - DB)

= [((f

BF1

- f

BS1

- DB1) + (f

BF2

- f

BS2

- DB2))/2] - PB1 - (AB2 - DB2)

BR1, BR2

Residue for duplicate blanks (g)

fBF

Final Blank Filter Bag (g)

fBS

Initial Blank Filter Bag (g)

Protein of Blank Filter Bag (g)

Ash of Blank Filter Bag (g)

Original wt of Diatomaceous Earth in Blank Filter Bag (g)

X 100

27. Determine the Protein content within the IDF and SDF residue.

See the “Protein Determination” section of this manual for recommended procedures.

28. Determine the Ash content within the IDF and SDF residue.

See the “Ash Determination” section of this manual for recommended procedures.

29. Calculate the % IDF value.

30. Calculate the % SDF value.

31. Calculate the % TDF value by adding the % IDF and % SDF values.

pg. 30 Rev E 12/2/14

Page 31

Operator’s Manual

Using a Bag Weigh Holder is critical to eliminate the effects of static electricity during the weighing process.

Because different balances have different sensitivities, the Bag Weigh Holder should be placed in the center of the balance for best results.

SDF Bag (longer bag with filter)

IDF Flow-thru Bag (shorter bag / no filter)

TDF Analysis (AOAC 991.43)

A TDF analysis directly measures the amount of TDF within a given sample without separately measuring the IDF and SDF fractions. TDF can be determined using multiple methods. This section describes the procedure for using the ANKOM

TDF

Dietary Fiber Analyzer to determine TDF based on the AOAC 991.43 method. This analysis requires an SDF filter bag (ANKOM DF-S) for the precipitation process and a non-filter IDF Flow-Thru bag

(ANKOM DF-FT) for the digestion process. When starting a new run, the ANKOM

TDF

Dietary Fiber Analyzer must have bags

installed at all stations (a total of twelve bags).

To perform a TDF analysis, follow the steps below.

1. Prepare chemicals and enzymes.

TDF

Dietary Fiber Analyzer for the AOAC 991.43, AACC 32-07.01, and NMKL 129,2003

2. De-fat (see "Appendix C – De-fatting Procedure" in this manual) and de-sugar your samples as

needed according to the AOAC 991.43, AACC 32-07.01, or NMKL 129,2003 methods.

3. Label the bags using a Solvent Resistant Marker.

4. Prepare for data collection.

5. Weigh filter bags.

Roll or fold each bag and place it in a tared Bag Weigh Holder. Place the Bag Weigh Holder in the center of a balance and record the weight.

Rev E 12/2/14 pg. 31

Page 32

Operator’s Manual

Although the instrument works in accordance with AACC and NMKL methods, the Touch Screen Display only refers to the AOAC methods.

←

Diagnostics

Touch Screen Display version: A.AA SPLat controller software version: vX.XX dd-mm-yy

Pump Tube

Test

Expert Mode

(OFF)

Faults

Service

Mode

Clear IDF /

SDF lines

Digestion

Times

Heater Test

Temperatures

Pressures

Line Charge

Motor Test

Valve Test

Select a Function

AOAC 991.43

IDF/SDF

AOAC 985.29

TDF

Diagnostics

AOAC 991.43

TDF

6. Weigh Diatomaceous Earth (DE).

DE is used during fiber analysis to enhance the flocculation and filtration of the SDF fraction. Place ca 1 g of DE in each of six tared and numbered tins and record the weights.

7. Weigh Samples.

Place 0.50.05 g of sample in each of six tared and numbered tins and record the weights.

8. Turn the instrument power on.

When you turn the power on, the instrument will run through an initialization process and the Control Panel will turn on.

9. On the Touch Screen Display, select the instrument function you would like to perform.

The Control Panel on the ANKOM you will press identified buttons on the Touch Screen Display and buttons below the screen.

When your instrument is initialized and ready to operate, the following screen will be displayed.

TDF

Dietary Fiber Analyzer uses Touch Screen technology. To operate the instrument

9.1 If this is the first time the instrument is being operated after being installed, or after sitting unused for a period of

time (see Appendix B), or after tubing was replaced, the lines must be charged to avoid a Vacuum Sensor Fault. To charge the lines follow the steps below.

9.1.1 Press the Diagnostics button on the “Select a Function” screen. The following screen will be displayed.

9.1.2 Press the Line Charge button. The following screen will be displayed.

pg. 32 Rev E 12/2/14

Page 33

Operator’s Manual

SDF Bags (and clamp bar D) installed?

AOAC 991.43 TDF

←

Select a Function

AOAC 991.43

IDF/SDF

AOAC 985.29

TDF

Diagnostics

AOAC 991.43

TDF

←

Line Charge

Amylase

Protease

AMG

HCl

EtOH78

EtOH95

Water

Buffer

ALL

9.1.3 Press the ALL button to charge all of the lines. Each button will change color as the associated line is being

charged. All lines are charged when all of the buttons return to their original green color.

9.1.4 Press the back button twice. The following “Select a Function” screen will be displayed again.

9.2 Press the AOAC 991.43 TDF button. The following screen will be displayed.

Rev E 12/2/14 pg. 33

Notice that the function you selected is now displayed in the top right corner of the screen and a back button is displayed in the top left corner of the screen.

Page 34

Operator’s Manual

As part of normal operation, solution from the IDF Flow-thru bag will flow into the SDF bag. Therefore, when installing the SDF filter bags it is very important to position them high enough vertically so that at least 20 mm (0.75 inches) of the bottom of the IDF Flow-thru bag can fit inside the top of the SDF bag.

Clamp Bar C

Clamp Bar D

Clamp Bar B

Clamp Bar A

10. Install SDF filter bags on the ANKOM

10.1 Remove Clamp Bars A, B, C, and D by lifting them off of the locator rods.

10.2 Gently pull the black SDF Delivery Nozzle out toward you.

TDF

Dietary Fiber Analyzer.

10.3 Place a labeled and weighed SDF bag up underneath the SDF Delivery Nozzle so that the Delivery Nozzle is

pg. 34 Rev E 12/2/14

Page 35

Operator’s Manual

When adding DE to the filter bags it is very important to keep it below the tip of the Delivery Nozzle so that the DE material can be properly rinsed.

Diatomaceous Earth

added in SDF Bags?

AOAC 991.43 TDF

←

Centering Lines

10.4 With the bag being held by the Delivery Nozzle, center it horizontally between the lines on the back part of

Clamp Bar C.

10.5 Repeat steps 10.2 – 10.4 for all six stations.

10.6 Re-install Clamp Bar D by setting it on the locator rods. Make sure the letter is on the top of the bar and the

rubber material is facing in toward the instrument.

10.7 Flatten the bags to remove any wrinkles.

10.8 With fingers away from the clamp bars, press the check mark () button on the Touch Screen Display to pinch

the bags. The following screen will be displayed.

11. Add DE to the SDF filter bags on the ANKOM

11.1 Open the top of the SDF bag and add a weigh tin of DE to the bag by folding the tin and then dipping it down into

the bag below the tip of the Delivery Nozzle.

TDF

Dietary Fiber Analyzer.

11.2 Rinse the tin with no more than 3 ml of Distilled Water to ensure complete transfer.

11.3 Repeat steps 11.1 – 11.2 for all six stations.

Rev E 12/2/14 pg. 35

Page 36

Operator’s Manual

After you confirm that the DE is added, the Clamp Bar D button on the Control Panel is disabled until the SDF process completes.

For proper mixing during the IDF process the IDF Flow-thru bags must be horizontally centered over the Heating Paddles and between the Centering Lines on the back part of Clamp Bar A.

IDF Flow-Thru Bags (and clamp bar B) installed?

AOAC 991.43 TDF

←

Centering Lines

Heating Paddle

11.4 Press the check mark () button on the Touch Screen Display. The following screen will be displayed.

12. Install IDF Flow-thru bags on the ANKOM

12.1 Gently pull the black IDF Delivery Nozzle out toward you.

12.2 Place a labeled IDF Flow-Thru bag up underneath the Delivery Nozzle so that the Delivery Nozzle is inside the

top part of the bag. Pull the bag up so that the top of the bag is about 35 mm (1.375 inches) above the top of Clamp Bar A and return the Delivery Nozzle to its original position. This will hold the back of the bag in place.

12.3 With the bag held by the Delivery Nozzle, center it horizontally between the Centering Lines on the back part of

Clamp Bar A.

TDF

Dietary Fiber Analyzer.

pg. 36 Rev E 12/2/14

Page 37

Operator’s Manual

When adding sample to the IDF Flow-thru bags it is very important to keep it below the tip of the Delivery Nozzle so that it can be properly rinsed.

Samples in IDF Flow-Thru Bags?

AOAC 991.43 TDF

←

20 mm (0.75 inches) of the IDF Flow-Thru bag inside the top of the SDF bag

Heating Paddle

12.4 Place at least 20 mm (0.75 inches) of the bottom of the IDF Flow-Thru bag inside the top of the SDF bag to allow

for the flow of solution into the SDF bag after the digestion process is complete.

12.5 Repeat steps 12.1 – 12.4 for all six stations.

12.6 Re-install Clamp Bar B by setting it on the locator rods. Make sure the letter is on the top of the bar and the

rubber material is facing in toward the instrument.

12.7 Flatten the IDF Flow-Thru bags to remove any wrinkles.

12.8 With fingers away from the clamp bars, press the check mark () button on the Touch Screen Display to pinch

the IDF Flow-Thru bags. The mixing pads will make contact with the bags when Clamp Bar B pinches the IDF Flow-Thru bags. The following screen will be displayed.

13. Add samples to the IDF Flow-thru bags.

13.1 Open the top of the IDF Flow-thru bag and transfer the sample from a weigh tin into the bag by folding the tin

and then dipping it down into the bag below the tip of the Delivery Nozzle.

13.2 Rinse the tin with no more than 3 ml of Distilled Water to ensure complete transfer.

Rev E 12/2/14 pg. 37

Page 38

Operator’s Manual

Hooking the bags in place will put a physical hole in the front of each SDF bag.

Front of SDF Bags hooked to clamp bar C?

AOAC 991.43 TDF

←

All remaining clamp bars installed?

AOAC 991.43 TDF

←

Hook on front part of Clamp Bar C

13.3 Repeat steps 13.1 – 13.2 for all IDF stations.

13.4 Re-install Clamp Bar A by setting it on the locator rods. Make sure the letter is on the top of the bar and the

rubber material is facing in toward the instrument.

13.5 Press the check mark () button on the Touch Screen Display. The following screen will be displayed.

13.6 Make sure that all clamp bars are installed with the letter on the top of the bar and the rubber material facing in

toward the instrument. Press the check mark () button on the Touch Screen Display. The following screen will be displayed.

14. Hook the front of each SDF bag in place.

14.1 Secure the SDF filter bags in place with the hooks located on the front part of Clamp Bar C.

14.2 Press the check mark () button on the Touch Screen Display. If you ran a method other than 991.43 prior to this

run, the following screen will be displayed.

pg. 38 Rev E 12/2/14

Page 39

Operator’s Manual

Do NOT leave the enzyme ports on the instrument open to the air or the enzymes in the valves may dry up and plug the ports.

Nitrogen supply ON?

AOAC 991.43 TDF

←



Fluid levels correct?

AOAC 991.43 TDF

←

Amylase -> Amylase AMG -> AMG HCL -> HCL Buffer -> Mes Tris Buffer

AOAC 991.43 TDF

←

Check supply containers

15. Verify that the fluid containers have the correct fluids for this procedure.

Verify that the fluid supply containers are configured according to the screen above and press the check mark () button. The following screen will be displayed.

16. Fill fluid containers.

To ensure that you have enough fluids to run a complete TDF procedure, you must begin with fluid levels above the Minimum Level lines on the chemical containers and at least 10 ml of each enzyme. Add fluids and enzymes as necessary.

With all fluid containers filled to the proper levels, press the check mark () button on the Touch Screen Display. The following screen will be displayed.

Rev E 12/2/14 pg. 39

Page 40

Operator’s Manual

during filtering if needed (see the “Status Screen” section of this manual for more

detail).

Check pH manually?

AOAC 991.43 TDF

←

Yes

AOAC 991.43 TDF

←



After release

After EtOH rinses

10.0

3.0

SDF Filter minutes OK?

(Press button with number to change)

17. Connect the Nitrogen Supply to the instrument and turn on.

Make sure the Nitrogen supply in your lab is connected to the instrument and turned on. The pressure gauges on the instrument should show 50-55 psi on the left and about 4 psi on the right.

Press the check mark () button on the Touch Screen Display. The following screen will be displayed.

18. Set filter times (in minutes).

When all of the times shown on the screen are what you want, press the check mark () button on the Touch Screen Display. The following screen will be displayed.

19. Set the Manual pH Check.

If you plan to check the pH after the required HCl is added (during the IDF process), press the Yes button on the screen above. Otherwise, press the No button. The following screen will be displayed.

pg. 40 Rev E 12/2/14

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Operator’s Manual

Testing pump tubing.

Please wait..........

Initializing lines…

Amylase

EtOH78

Protease

EtOH95

AMG

Water

HCL

Buffer

Vacuum sensor:

AAAA

psi

Press START to begin

AOAC 991.43 TDF

←

START

20. Start the instrument.

The instrument is now completely set up and ready to automatically run the TDF procedure. Press the START button to begin. At the beginning of each new run, the instrument automatically runs a tube integrity test.

For the first run after a power-up cycle, or for the first run after the instrument has been idle for twelve hours, the following screen will be displayed.

If the fluid lines are already charged properly, or when the line charge is complete, the following screen will be displayed.

When the tube integrity check is complete, the instrument will automatically execute the TDF procedure stopping only for faults, aborts, and manual pH measurement (if enabled). The Status screen will show actions and faults as they occur during the automatic operation.

Rev E 12/2/14 pg. 41

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Operator’s Manual

When measuring pH use a probe that can be easily rinsed (with Distilled Water) to avoid loss of sample. If you add acid or base to adjust the pH, you must mix the solution in order to get an accurate pH reading. To manually mix the solution, press the outside of the bag with your fingers just above the clamp bar multiple times.

To rinse the SDF filter bags with acetone using the ANKOM TDF51 Rinse Stand, follow the steps below.

23.1 Remove the SDF bags from the instrument.

23.2 Place the bags on the Rinse Stand by sliding the back part of the bag under the pinch

mechanism. Keep the top of the bag open.

23.3 Using a wash bottle, squirt acetone into each bag, making sure that all residue on the

clear polypropylene surfaces is rinsed down into the filter.

23.4 Repeat step 23.3 so that each bag gets rinsed a total of two times.

23.5 Allow acetone to evaporate from the bags.

Hazardous Materials – Do NOT heat seal or place bags in an oven until all

acetone has evaporated.

Status

AOAC 991.43 TDF

Overall Progress

Action in progress

IDF

Time remaining:

Amylase/AMG Phase

XX:XX:XX

Time remaining: XX:XX:XX

SDF

Time remaining:

XX:XX:XX

pH required?

Display

Temps/Pressures YES

IDF

ABORT

PAUSE

SDF

ABORT

Check pH

then press continue

xx:xx:xx

Continue

21. Manually measure pH.

below, and make a sound to remind you to manually measure pH and adjust if necessary.

When you have completed the pH measurement process, press the Continue button on the Status screen. You will see “COMPLETE” next to “pH required?” in the upper left corner of the screen.

22. Make sure that each sample is completely rinsed down into the SDF filter bags (as needed).

Since the IDF Flow-thru bags are not weighed before and after the process, it is important to make sure that each sample gets rinsed down into the SDF filter bag when the digestion process is complete.

If you notice sample sticking to an IDF Flow-thru bag, use 78% EtOH in a wash bottle to rinse as needed.

23. Rinse the SDF filter bags with acetone.

After the instrument has completed a TDF process the TDF residue that has been collected in the SDF bag must be manually rinsed with acetone. It is recommended that you use an ANKOM TDF51 Rinse Stand (sold separately) for rinsing filter bags with acetone.

pg. 42 Rev E 12/2/14

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23.6 With your Heat Sealer set between 3 and 4 (settings may vary depending on the

heat sealer and the power source), seal each bag just above the filter to keep all residue contained to the filter area while handling the bags.

23.7 Place each bag in the Drying Rack.

% TDF

[(R1 + R2)/2] - P - A - B

(M1 + M2)/2

[(( fF1 - fS1 - D1) + ( fF2 - fS2 - D2))/2] - P1 - (A2 - D2) - B

(M1 + M2)/2

Where:

M1, M2

Original wt of duplicate samples adjusted for pre-treatment fat and sugar losses (g)

R1, R2

Residue for duplicate samples (g)

Final Filter Bag with residue (g)

Initial Filter Bag (g)

D =

Original wt of Diatomaceous Earth (g)

P =

Protein of residue and bag (g)

Ash of residue and bag (g)

Blank (g)

= [(BR1 + BR2)/2] - PB - (AB - DB)

= [((f

BF1

- f

BS1

- DB1) + (f

BF2

- f

BS2

- DB2))/2] - PB1 - (AB2 - DB2)

BR1, BR2

Residue for duplicate blanks (g)

fBF

Final Blank Filter Bag (g)

fBS

Initial Blank Filter Bag (g)

Protein of Blank Filter Bag (g)

Ash of Blank Filter Bag (g)

Original wt of Diatomaceous Earth in Blank Filter Bag (g)

X 100

24. Dry the SDF filter bags.

24.1 Make sure your oven is 105°C at the location where the bags will be placed.

24.2 Place the Drying Rack with the filter bags in the oven and dry to constant weight (about 90 minutes).

24.3 When dry, remove all SDF filter bags from the oven and place them in a desiccant pouch to cool.

25. Weigh the TDF residue.

25.1 Remove one SDF filter bag from the desiccant pouch.

25.2 Roll or fold the bag, place it in a tared Bag Weigh Holder (BWH), and place the BWH in the center of a balance.

25.3 Record the weight of the filter bag.

25.4 Repeat steps 25.1 – 25.3 for each SDF filter bag used in the fiber analysis.

26. Determine the Protein content within the TDF residue.

See the “Protein Determination” section of this manual for recommended procedures.

27. Determine the Ash content within the TDF residue.

See the “Ash Determination” section of this manual for recommended procedures.

28. Calculate the % TDF value.

Rev E 12/2/14 pg. 43

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pg. 44 Rev E 12/2/14

Page 45

Operator’s Manual

Using a Bag Weigh Holder is critical to eliminate the effects of static electricity during the weighing process.

Because different balances have different sensitivities, the Bag Weigh Holder should be placed in the center of the balance for best results.

SDF Bag (longer bag with filter)

IDF Flow-thru Bag (shorter bag / no filter)

TDF Analysis (AOAC 985.29)

TDF

Dietary Fiber Analyzer to determine TDF based on the AOAC 985.29 method. This analysis requires an SDF filter bag (ANKOM DF-S) for the precipitation process and a non-filter IDF Flow-Thru bag

(ANKOM DF-FT) for the digestion process. When starting a new run, the ANKOM

TDF

Dietary Fiber Analyzer must have bags

installed at all stations (a total of twelve bags).

To perform a TDF analysis, follow the steps below.

1. Prepare chemicals and enzymes.

When using the ANKOM referred to therein. See Appendix A of this manual for the list of chemicals and enzymes and the instructions for how to prepare them for use in this instrument.

TDF

Dietary Fiber Analyzer for the AOAC 985.29 method use the chemicals and enzymes

2. De-fat (see "Appendix C – De-fatting Procedure" in this manual) and de-sugar your samples as

needed according to the AOAC 985.29 method.

3. Label the bags using a Solvent Resistant Marker.

4. Prepare for data collection.

You will need a place to store the data collected during this analysis. For your convenience a CD titled “Calculation Templates” is included with the instrument. This CD has TDF Data Spreadsheets that can be used for this analysis. Please read the “Instructions” tab in the MS-Excel file for information about the spreadsheets.

5. Weigh filter bags.

Roll or fold each bag and place it in a tared Bag Weigh Holder. Place the Bag Weigh Holder in the center of a balance and record the weight.

Rev E 12/2/14 pg. 45

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Operator’s Manual

Although the instrument works in accordance with AACC and NMKL methods, the Touch Screen Display only refers to the AOAC methods.

←

Diagnostics

Touch Screen Display version: A.AA SPLat controller software version: vX.XX dd-mm-yy

Pump Tube

Test

Expert Mode

(OFF)

Faults

Service

Mode

Clear IDF /

SDF lines

Digestion

Times

Heater Test

Temperatures

Pressures

Line Charge

Motor Test

Valve Test

Select a Function

AOAC 991.43

IDF/SDF

AOAC 985.29

TDF

Diagnostics

AOAC 991.43

TDF

6. Weigh Diatomaceous Earth (DE).

DE is used during fiber analysis to enhance the flocculation and filtration of the SDF fraction. Place ca 1 g of DE in each of six tared and numbered tins and record the weights.

7. Weigh Samples.

Place 0.50.05 g of sample in each of six tared and numbered tins and record the weights.

8. Turn the instrument power on.

When you turn the power on, the instrument will run through an initialization process and the Control Panel will turn on.

9. On the Touch Screen Display, select the instrument function you would like to perform.

The Control Panel on the ANKOM you will press identified buttons on the Touch Screen Display and buttons below the screen.

When your instrument is initialized and ready to operate, the following screen will be displayed.

TDF

Dietary Fiber Analyzer uses Touch Screen technology. To operate the instrument

9.1 If this is the first time the instrument is being operated after being installed, or after sitting unused for a period of

time (see Appendix B), or after tubing was replaced the lines must be charged to avoid a Vacuum Sensor Fault. To charge the lines follow the steps below.

9.1.1 Press the Diagnostics button on the “Select a Function” screen. The following screen will be displayed.

9.1.2 Press the Line Charge button. The following screen will be displayed.

pg. 46 Rev E 12/2/14

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Operator’s Manual

SDF Bags (and clamp bar D) installed?

AOAC 985.29 TDF

←

Select a Function

AOAC 991.43

IDF/SDF

AOAC 985.29

TDF

Diagnostics

AOAC 991.43

TDF

←

Line Charge

Amylase

Protease

AMG

HCl

EtOH78

EtOH95

Water

Buffer

ALL

9.1.3 Press the ALL button to charge all of the lines. Each button will change color as the associated line is being

charged. All lines are charged when all of the buttons return to their original green color.

9.1.4 Press the back button twice. The following “Select a Function” screen will be displayed again.

9.2 Press the AOAC 985.29 TDF button. The following screen will be displayed.

Rev E 12/2/14 pg. 47

Notice that the function you selected is now displayed in the top right corner of the screen and a back button is displayed in the top left corner of the screen.

Page 48

Operator’s Manual

Clamp Bar C

Clamp Bar D

Clamp Bar B

Clamp Bar A

10. Install SDF filter bags on the ANKOM

10.1 Remove Clamp Bars A, B, C, and D by lifting them off of the locator rods.

10.2 Gently pull the black SDF Delivery Nozzle out toward you.

TDF

Dietary Fiber Analyzer.

10.3 Place a labeled and weighed SDF bag up underneath the SDF Delivery Nozzle so that the Delivery Nozzle is

pg. 48 Rev E 12/2/14

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Operator’s Manual

When adding DE to the filter bags it is very important to keep it below the tip of the Delivery Nozzle so that the DE material can be properly rinsed.

Diatomaceous Earth

added in SDF Bags?

AOAC 985.29 TDF

←

Centering Lines

10.4 With the bag being held by the Delivery Nozzle, center it horizontally between the lines on the back part of

Clamp Bar C.

10.5 Repeat steps 10.2 – 10.4 for all six stations.

10.6 Re-install Clamp Bar D by setting it on the locator rods. Make sure the letter is on the top of the bar and the

rubber material is facing in toward the instrument.

10.7 Flatten the bags to remove any wrinkles.

10.8 With fingers away from the clamp bars, press the check mark () button on the Touch Screen Display to pinch

the bags. The following screen will be displayed.

11. Add DE to the SDF filter bags on the ANKOM

11.1 Open the top of the SDF bag and add a weigh tin of DE to the bag by folding the tin and then dipping it down into

the bag below the tip of the Delivery Nozzle.

TDF

Dietary Fiber Analyzer.

11.2 Rinse the tin with no more than 3 ml of Distilled Water to ensure complete transfer.

11.3 Repeat steps 11.1 – 11.2 for all six stations.

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After you confirm that the DE is added, the Clamp Bar D button on the Control Panel is disabled until the SDF process completes.

For proper mixing during the IDF process the IDF Flow-thru bags must be horizontally centered over the Heating Paddles and between the Centering Lines on the back part of Clamp Bar A.

IDF Flow-Thru Bags (and clamp bar B) installed?

AOAC 985.29 TDF

←

Centering Lines

Heating Paddle

11.4 Press the check mark () button on the Touch Screen Display. The following screen will be displayed.

12. Install IDF Flow-thru bags on the ANKOM

12.1 Gently pull the black IDF Delivery Nozzle out toward you.

12.2 Place a labeled IDF Flow-Thru bag up underneath the Delivery Nozzle so that the Delivery Nozzle is inside the

12.3 With the bag held by the Delivery Nozzle, center it horizontally between the Centering Lines on the back part of

Clamp Bar A.

TDF

Dietary Fiber Analyzer.

pg. 50 Rev E 12/2/14

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Operator’s Manual

When adding sample to the IDF Flow-thru bags it is very important to keep it below the tip of the Delivery Nozzle so that it can be properly rinsed.

Samples in IDF Flow-Thru Bags?

AOAC 985.29 TDF

←

20 mm (0.75 inches) of the IDF Flow-Thru bag inside the top of the SDF bag

Heating Paddle

12.4 Place at least 20 mm (0.75 inches) of the bottom of the IDF Flow-Thru bag inside the top of the SDF bag to allow

for the flow of solution into the SDF bag after the digestion process is complete.

12.5 Repeat steps 12.1 – 12.4 for all six stations.

12.6 Re-install Clamp Bar B by setting it on the locator rods. Make sure the letter is on the top of the bar and the

rubber material is facing in toward the instrument.

12.7 Flatten the IDF Flow-Thru bags to remove any wrinkles.

12.8 With fingers away from the clamp bars, press the check mark () button on the Touch Screen Display to pinch

the IDF Flow-Thru bags. The mixing pads will make contact with the bags when Clamp Bar B pinches the IDF Flow-Thru bags. The following screen will be displayed.

13. Add samples to the IDF Flow-thru bags.

13.1 Open the top of the IDF Flow-thru bag and transfer the sample from a weigh tin into the bag by folding the tin

and then dipping it down into the bag below the tip of the Delivery Nozzle.

13.2 Rinse the tin with no more than 3 ml of Distilled Water to ensure complete transfer.

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Hooking the bags in place will put a physical hole in the front of each SDF bag.

Front of SDF Bags hooked to clamp bar C?

AOAC 985.29 TDF

←

All remaining clamp bars installed?

AOAC 985.29 TDF

←

Hook on front part of Clamp Bar C

13.3 Repeat steps 13.1 – 13.2 for all IDF stations.

13.4 Re-install Clamp Bar A by setting it on the locator rods. Make sure the letter is on the top of the bar and the

rubber material is facing in toward the instrument.

13.5 Press the check mark () button on the Touch Screen Display. The following screen will be displayed.

13.6 Make sure that all clamp bars are installed with the letter on the top of the bar and the rubber material facing in

toward the instrument. Press the check mark () button on the Touch Screen Display. The following screen will be displayed.

14. Hook the front of each SDF bag in place.

14.1 Secure the SDF filter bags in place with the hooks located on the front part of Clamp Bar C.

14.2 Press the check mark () button on the Touch Screen Display. If you ran a method other than 985.29 prior to this

run, the following screen will be displayed.

pg. 52 Rev E 12/2/14

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Operator’s Manual

Do NOT leave the enzyme ports on the instrument open to the air or the enzymes in the valves may dry up and plug the ports.

Nitrogen supply ON?

AOAC 985.29 TDF

←



Fluid levels correct?

AOAC 985.29 TDF

←

Amylase -> Amylase AMG -> AMG HCL -> HCL Buffer -> Phosphate Buffer

AOAC 985.29 TDF

←

Check supply containers

15. Verify that the fluid containers have the correct fluids for this procedure.

Verify that the fluid supply containers are configured according to the screen above and press the check mark () button. The following screen will be displayed.

16. Fill fluid containers.

With all fluid containers filled to the proper levels, press the check mark () button on the Touch Screen Display. The following screen will be displayed.

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Operator’s Manual

during filtering if needed (see the “Status Screen” section of this manual for more

detail).

Check pH manually?

AOAC 985.29 TDF

←

Yes

AOAC 985.29 TDF

←



After release

After EtOH rinses

10.0

3.0

SDF Filter minutes OK?

(Press button with number to change)

17. Connect the Nitrogen Supply to the instrument and turn on.

Make sure the Nitrogen supply in your lab is connected to the instrument and turned on. The pressure gauges on the instrument should show 50-55 psi on the left and about 4 psi on the right.

Press the check mark () button on the Touch Screen Display. The following screen will be displayed.

18. Set filter times (in minutes).

When all of the times shown on the screen are what you want, press the check mark () button on the Touch Screen Display. The following screen will be displayed.

19. Set the manual pH check for after the Protease digestion.

When running the AOAC 985.29 method, the instrument will always stop just before the Protease digestion (after the bags have cooled to the appropriate temperature) and ask you to adjust pH.

If you plan to also check/adjust the pH after the Protease digestion, press the Yes button on the screen above. Otherwise, press the No button. The following screen will be displayed.

pg. 54 Rev E 12/2/14

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Operator’s Manual

Testing pump tubing.

Please wait..........

Initializing lines…

Amylase

EtOH78

Protease

EtOH95

AMG

Water

HCL

Buffer

Vacuum sensor:

AAAA

psi

Press START to begin

AOAC 985.29 TDF

←

START

20. Start the instrument.

For the first run after a power-up cycle, or for the first run after the instrument has been idle for twelve hours, the following screen will be displayed.

If the fluid lines are already charged properly, or when the line charge is complete, the following screen will be displayed.

When the tube integrity check is complete, the instrument will automatically execute the TDF procedure stopping only for faults, aborts, and manual pH adjustment. The Status screen will show actions and faults as they occur during the automatic operation.

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When measuring/adjusting pH use a probe that can be easily rinsed (with Distilled Water) to avoid loss of sample. If you add acid or base to adjust the pH, you must mix the solution in order to get an accurate pH reading. To manually mix the solution, press the outside of the bag with your fingers just above the clamp bar multiple times.

Status

AOAC 985.29 TDF

Overall Progress

Action in progress

IDF

Time remaining:

Amylase/AMG Phase

XX:XX:XX

Time remaining: XX:XX:XX

SDF

Time remaining:

XX:XX:XX

pH required?

Display

Temps/Pressures YES

IDF

ABORT

PAUSE

SDF

ABORT

Check pH

then press continue

xx:xx:xx

Continue

AOAC 985.29 pH adjustment

Continue

Adjust pH to 7.5 +/- 0.2 by adding NaOH solution.

Press "Continue" when complete.

21. Adjust pH before the Protease digestion.

Just before the Protease digestion (after the bags have cooled to the appropriate temperature) the instrument will stop, open Clamp Bar A, display the screen below, and make a sound to remind you to manually adjust pH.

After adjusting the pH, press the Continue button.

22. Manually measure/adjust pH after the Protease digestion.

below, and make a sound to remind you to manually measure pH and adjust if necessary.

When you have completed the pH measurement process, press the Continue button on the Status screen. You will see “COMPLETE” next to “pH required?” in the upper left corner of the screen.

23. Make sure that each sample is completely rinsed down into the SDF filter bags (as needed).

If you notice sample sticking to an IDF Flow-thru bag, use 78% EtOH in a wash bottle to rinse as needed.

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Operator’s Manual

24.1 Remove the SDF bags from the instrument.

24.2 Place the bags on the Rinse Stand by sliding the back part of the bag under the

pinch mechanism. Keep the top of the bag open.

24.3 Using a wash bottle, squirt acetone into each bag, making sure that all residue on

the clear polypropylene surfaces is rinsed down into the filter.

24.4 Repeat step 24.3 so that each bag gets rinsed a total of two times.

24.5 Allow acetone to evaporate from the bags.

Hazardous Materials – Do NOT heat seal or place bags in an oven until all

acetone has evaporated.

24.6 With your Heat Sealer set between 3 and 4 (settings may vary depending on

the heat sealer and the power source), seal each bag just above the filter to keep all residue contained to the filter area while handling the bags.

24.7 Place each bag in the Drying Rack.

24. Rinse the SDF filter bags with acetone.

To rinse the SDF filter bags with acetone using the ANKOM TDF51 Rinse Stand, follow the steps below.

25. Dry the SDF filter bags.

25.1 Make sure your oven is 105°C at the location where the bags will be placed.

25.2 Place the Drying Rack with the filter bags in the oven and dry to constant weight (about 90 minutes).

25.3 When dry, remove all SDF filter bags from the oven and place them in a desiccant pouch to cool.

26. Weigh the TDF residue.

26.1 Remove one SDF filter bag from the desiccant pouch.

26.2 Roll or fold the bag, place it in a tared Bag Weigh Holder (BWH), and place the BWH in the center of a balance.

26.3 Record the weight of the filter bag.

26.4 Repeat steps 26.1 – 26.3 for each SDF filter bag used in the fiber analysis.

27. Determine the Protein content within the TDF residue.

See the “Protein Determination” section of this manual for recommended procedures.

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Operator’s Manual

% TDF

[(R1 + R2)/2] - P - A - B

(M1 + M2)/2

[(( fF1 - fS1 - D1) + ( fF2 - fS2 - D2))/2] - P1 - (A2 - D2) - B

(M1 + M2)/2

Where:

M1, M2

Original wt of duplicate samples adjusted for pre-treatment fat and sugar losses (g)

R1, R2

Residue for duplicate samples (g)

Final Filter Bag with residue (g)

Initial Filter Bag (g)

D =

Original wt of Diatomaceous Earth (g)

P =

Protein of residue and bag (g)

Ash of residue and bag (g)

Blank (g)

= [(BR1 + BR2)/2] - PB - (AB - DB)

= [((f

BF1

- f

BS1

- DB1) + (f

BF2

- f

BS2

- DB2))/2] - PB1 - (AB2 - DB2)

BR1, BR2

Residue for duplicate blanks (g)

fBF

Final Blank Filter Bag (g)

fBS

Initial Blank Filter Bag (g)

Protein of Blank Filter Bag (g)

Ash of Blank Filter Bag (g)

Original wt of Diatomaceous Earth in Blank Filter Bag (g)

X 100

28. Determine the Ash content within the TDF residue.

See the “Ash Determination” section of this manual for recommended procedures.

29. Calculate the % TDF value.

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AOAC method 991.43 suggests that the Kjeldahl method be used for determining protein content. Since a large amount of acid is used in the Kjeldahl digestion process, make sure you use enough base in the Kjeldahl distillation process.

Cut the filter bag just above the seal.

Cut off the polypropylene skirt that covers the filter (both sides) being careful to keep the filter sealed and intact. This minimizes the acid needed to digest the filter bag during the Kjeldahl procedure.

Fold the remainder of the filter bag in thirds from side to side; then fold from top to bottom.

With your Heat Sealer set to “6” (setting may vary depending on heat sealer and power source), place the exposed polypropylene material from your final fold down, and press the Heat Sealer arm down for 3 to 4 seconds to seal each bag. Repeat if necessary.

Label the folded, sealed filter bag.

Repeat steps 1 – 5 for all IDF bags that will be used for determining protein content.

Run a Kjeldahl procedure on each filter bag using 17 ml of H2SO4, 10 g of K2SO4, and 30 mg of CuSO45H2O in a system with 250 ml tubes that have a 42 mm diameter. Pellets can be used if an equivalent amount of K2SO4 is added. To slow down the rate of foaming during the digestion process, ramp up the temperature slowly.

Protein Determination

IDF – Kjeldahl Method

To determine the protein content within the IDF residue, follow the procedure below.

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Cut the filter bag 8 -10 mm above the filter.

Fold the remainder of the filter bag in thirds from side to side.

With your Heat Sealer set to “6” (setting may vary depending on heat sealer and power source), press the arm down for 3 to 4 seconds to seal the polypropylene edge of the filter bag. Repeat if necessary.

Label the folded, sealed filter.

Repeat steps 1 – 4 for all SDF bags that will be used for determining protein content.

Run a Kjeldahl procedure on each filter bag using 27 ml of H2SO4, 13 g of K2SO4, and 45 mg of CuSO45H2O in a system with 250 ml tubes that have a 42 mm diameter. Pellets can be used if an equivalent amount of K2SO4 is added. To slow down the rate of foaming during the digestion process, ramp up the temperature slowly.

SDF / TDF – Kjeldahl Method

To determine the protein content within the SDF or TDF residue, follow the procedure below.

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ANKOM Technology recommends that you use ceramic crucibles with fitted covers to avoid cross contamination or loss of ash. Fold the filter bags so that they easily fit into the crucibles.

Starting at the bottom of the filter part of the bag, fold the bag upward, leaving at least 10 mm at the top of the clear polypropylene that is not folded. Then fold the bag in thirds from side to side.

With your Heat Sealer set to “6” (setting may vary depending on heat sealer and power source), press the Heat Sealer arm down for 3 to 4 seconds to seal the clear polypropylene part of the bag just above the white filter material.

Label the folded, sealed filter bag and place it in a labeled ashing crucible.

Repeat steps 1 – 3 for all IDF bags to be ashed.

Place covered ashing crucibles with filter bags in a furnace at 600°C for 3 hours.

After the 3 hour furnace time, place the ashed crucibles in desiccator(s) and allow them to cool.

Remove cooled crucibles from the desiccator(s) and record their weights.

10 mm of clear polypropylene

Ash Determination

IDF

To determine the ash content within the IDF residue, follow the procedure below.

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During a TDF procedure there is no residue captured in the IDF Flow-Thru bag. Therefore, the ash procedure is only required for the SDF filter bag.

DE used in the precipitation process can sometimes be difficult to fully rinse from the walls of the polypropylene bags. Therefore, ANKOM Technology recommends that the entire SDF bag be ashed.

Set your Heat Sealer to “6” (setting may vary depending on heat sealer and power source) and press the Heat Sealer arm down for 3 to 4 seconds to seal the unfolded polypropylene to the folded part of the bag.

Label the folded, sealed filter bag and place it in a labeled ashing crucible.

Repeat steps 1 – 3 for all SDF bags to be ashed.

Place covered ashing crucibles with filter bags in a furnace at 600°C for 3 hours.

After the 3 hour furnace time, place the ashed crucibles in desiccator(s) and allow them to cool.

Remove cooled crucibles from the desiccator(s) and record their weights.

10 mm of clear polypropylene

SDF / TDF

To determine the ash content within the SDF / TDF residue, follow the procedure below.

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When starting a new run, the ANKOM

TDF

Dietary Fiber Analyzer must have bags installed at all stations (a total of twelve bags). If you are starting a digestion process while a precipitation process is running, you must install IDF or IDF Flow- Thru bags at all six IDF stations.

Select a Function

AOAC 991.43

IDF/SDF

AOAC 985.29

TDF

AOAC 991.43

TDF

←

Vacuum sensor:

AAAA

psi

Status

Overall Progress

Action in progress

IDF

Time remaining:

Process Complete

XX:XX:XX

SDF

Time remaining:

Precipitating.........

XX:XX:XX

Time remaining: XX:XX:XX

pH required?

Display

Temps/Pressures Yes

Method & Procedure

Start New

Procedure?

Vacuum sensor:

AAAA

psi

PAUSE

SDF

ABORT

Productivity Enhancement

Start a new procedure while a previous one is in progress

To enhance the productivity of your instrument, you can begin the digestion process (run in the IDF part of the instrument) of a new procedure (IDF/SDF or TDF) while the precipitation process (run in the SDF part of the instrument) of a previous procedure (IDF/SDF or TDF) is still running.

When a digestion process completes, the following screen is displayed showing that the process in the top bag is complete. The Method & Procedure shown in the top right corner will be the one that is currently in progress in the bottom bag.

At this time you can remove the IDF or IDF Flow-Thru bag from the instrument; then press the Start New Procedure button on the screen. The following screen will be displayed.

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Because an SDF process from the previous run is still in progress, you can leave the bottom of the IDF or IDF Flow-Thru bags outside of the SDF bags for now. However, you must REMEMBER to place the bottom of the IDF or IDF Flow- Thru bags in the top of the SDF bags after new SDF bags are installed.

After the pump tube testing is complete, the Status screen will be displayed.

The procedure shown in the top right corner of the Status screen reflects the procedure that was most recently started. Therefore, if you ran an IDF/SDF procedure, and then started a TDF procedure before the precipitation process for the previous IDF/SDF procedure completed, the top right corner of the display will show “TDF” even though the IDF/SDF procedure is not fully complete.

The material from the IDF or IDF Flow-Thru bags will not empty into the SDF bags until you verify that new SDF bags have been installed.

Testing pump tubing.

Please wait..........

Press START to begin

Method & Procedure

←

START

To start a new assay immediately after an IDF process completes, follow the steps below.

1. Prepare samples and configure the instrument for a new digestion.

Refer to the IDF/SDF Analysis and TDF Analysis sections of this manual, following all steps except for those that deal with the precipitation process (SDF). This will include interaction with the Touch Screen Display to verify the instrument set-up (as shown in the IDF/SDF Analysis and TDF Analysis sections).

After you confirm your set-up, the following screen is displayed. The Method & Procedure shown in the top right corner will be the one that you just started in the top bag.

2. Perform the digestion process using the ANKOM

To perform the new digestion process, press the START button to begin. The following screen will be displayed.

TDF

Dietary Fiber Analyzer.

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Remember to place the bottom of the IDF or IDF Flow-thru bags in the top of the SDF bags when installing the new SDF bags.

Vacuum sensor:

AAAA

psi

Overall Progress

Action in progress

IDF

Time remaining:

XX:XX:XX

Time remaining: XX:XX:XX

SDF

Time remaining:

XX:XX:XX

pH required?

Display

Temps/Pressures Yes

Method & Procedure

Vacuum sensor:

AAAA

psi

PAUSE

Vacuum sensor:

AAAA

psi

Overall Progress

Action in progress

IDF

Time remaining:

XX:XX:XX

Time remaining: XX:XX:XX

SDF

Time remaining:

Process

XX:XX:XX

Complete

pH required?

Display

Temps/Pressures Yes

Method & Procedure

Vacuum sensor:

AAAA

psi

PAUSE

Press to

continue SDF

3. Install new SDF bags on the ANKOM

When an SDF process completes while an IDF process is in progress, the following screen is displayed. The Method & Procedure shown in the top right corner is the one that is running in the top bag.

In the “Action in progress” box for the SDF process you will see a Press to continue SDF button. Press that button and follow the instructions on the Touch Screen Display.

TDF

Dietary Fiber Analyzer and continue the run.

After you confirm your set-up, the following Status screen is displayed. The Method & Procedure shown in the top right corner is the one that is running in the top bag.

The instrument is now ready to run an SDF process that will be automatically initiated by the internal computer at the correct time.

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You must be careful while in Expert Mode or you could cause problems!

The IDF/SDF and TDF analyses both start filling the SDF bag with EtOH about an hour after the IDF process starts. Therefore, if you start the IDF process, and wait more than an hour to properly install your SDF bags, the instrument will deliver EtOH to the empty bottom stations, wasting EtOH, and ruining a run.

While in Expert Mode you will use the buttons on the Control Panel just below the Touch Screen Display to operate the clamp bars.

←

Diagnostics

Touch Screen Display version: A.AA SPLat controller software version: vX.XX dd-mm-yy

Pump Tube

Test

Expert Mode

(OFF)

Faults

Service

Mode

Clear IDF /

SDF lines

Digestion

Times

Heater Test

Temperatures

Pressures

Line Charge

Motor Test

Valve Test

Select a Function

AOAC 991.43

IDF/SDF

AOAC 985.29

TDF

Diagnostics

AOAC 991.43

TDF

Expert Mode – save some set-up time

At the beginning of each new run, the instrument gives the user step-by-step instructions for set-up. These instructions are provided to make sure that all important steps are done before starting a run. After gaining experience with the instrument by running it over time, a user can choose to skip all set-up instructions except for “Check pH manually?” and “Filter minutes OK?” by enabling Expert Mode through Diagnostics.

The primary benefits of skipping most of the set-up instructions are:

 Time saved at the beginning of a run.  The ability to change the order of set-up. For example, a user can install the IDF bags first, start the IDF process

running, and then install the SDF bags.

To enter Expert Mode, follow the steps below.

1. From the Select a Function screen below, press the Diagnostics button.

The following screen will be displayed.

2. Press the Expert Mode button until it says “ON.”

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If you start a TDF procedure while you are still running the SDF process from an IDF/SDF procedure, “TDF” is displayed in this box even though an SDF process is in progress.

This button is only displayed when an ABORT operation is allowed.

Overall Progress

Action in progress

5 7 8

pH required?

13 1 15

Vacuum sensor:

psi

Status

Status Screen

The screen below shows the primary sections of the instrument Status screen when running an assay. This screen is NOT displayed when running Diagnostics.

The Status screen includes sections that present information that cannot be changed by the user, along with buttons that allow the user to modify the current operations or get additional information.

For explanation purposes, we have placed a number in each section of the screen and white borders around boxes 1 and 2. Your computer screen will NOT show these numbers nor will it show the borders around boxes 1 and 2.

1. pH check

Box #1 contains information about the manual pH check. One of the questions you are asked before starting the automated procedure is: “Check pH manually?” If you answered “Yes” to this question, the word “YES” is displayed in box #1 until the pH has been checked, at which time the word “COMPLETE” is displayed. Otherwise, the word “NO” is displayed in this box.

2. Function and Procedure

Box #2 displays “AOAC 991.43” along with the most recent procedure that you have started (either “IDF/SDF” or “TDF”). For example, “AOAC 991.43 IDF/SDF” is displayed when the most recent procedure started is an IDF/SDF.

3. Top process – Name

Box #3 always displays “IDF” to represent an IDF process within an IDF/SDF procedure or the digestion process within a TDF procedure.

4. Top process – Progress

Box #4 displays the actual time remaining for the top process.

5. Top process – Action

Box #5 displays different detailed actions as they occur during the process that is currently running in the top section of the instrument.

6. Top process – ABORT

When pressed, button #6 aborts the process currently running in the top section of the instrument.

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While paused, the instrument will continue the mechanical operation of the current step. For example, if you pause the instrument while mixing is occurring in the IDF process, the mixing paddles will continue to function even though the process timer has paused.

This button is only displayed when an ABORT operation is allowed.

←

Display

Temperatures

Display

Pressures

7. Bottom process – Name

Box #7 always displays “SDF” to represent an SDF process within an IDF/SDF procedure or the precipitation process within a TDF procedure.

8. Bottom process – Progress

Box #8 displays the actual time remaining for the bottom process.

9. Bottom process – Action

Box #9 displays different detailed actions as they occur during the process that is currently running in the bottom section of the instrument. When completing an SDF process while an IDF process is in progress, the “Press to continue SDF” button will appear in this box.

10. PAUSE

When pressed, button #10 pauses the timers for all processes currently in progress. This will stop the next process from starting. While paused, the button will read “CONT.” To continue the operation, press the “CONT” button.

11. Bottom process – ABORT

When pressed, button #11 aborts the process currently running in the bottom section of the instrument.

12. FAULT

Button #12 is displayed if a fault occurs within the instrument. When button #12 is pressed, the Fault Screen is displayed. This screen has information about all faults that have not been cleared. See the Fault Handling section of this manual for more information.

13. Display Temps/Pressures

The instrument allows the user to view temperature and pressure readings during an assay. When button #13 is pressed, the following screen is displayed.

When the Display Temperatures button is pressed, the following screen is displayed showing the temperatures of the Inline Heater and all six Heating Paddles.

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If you mistakenly increase your filter time using the Filter time increment button, you can use the Filter bypass to stop the filter process before the additional filter time occurs.

Pressure Monitor (psi)

←

xx.x

Line 1

xx.x

Line 2

xx.x

Line 3

xx.x

Line 4

xx.x

Line 5

xx.x

Line 6

xx.x

Input Pressure

xx.x

Vacuum Sensor

Temperature Monitor (°C)

←

Paddle 1

Paddle 2

Paddle 3

Paddle 4

Paddle 5

Paddle 6

In-line Heater

The values shown on this screen will dynamically change as the temperatures change. When the Display Pressures button is pressed, the following screen is displayed.

The readings shown on this screen for the input pressure and the lines to all six stations represent the absolute pressure. These values will dynamically change as the pressures change.

14. Start new procedures

Button #14 is displayed when a top process completes. At that time, the button reads: “Start New Procedure?” Button #14

is also displayed when a bottom process completes and no top process is in progress. At that time, the button reads: “Main

Menu.” When button #14 is pressed, the appropriate screen for starting a new procedure is displayed.

15. Filter time increment

Button #15 is displayed when a top or bottom process begins its filter action. At that time, the button reads: “Filter Time +.” When button #15 is pressed, the remaining filter time is increased by one minute for every time the button is pressed up to a limit of 54 additional minutes.

16. Filter bypass or Precipitation bypass

Button #16 is displayed when a top or bottom process begins its filter action or when a bottom process begins its precipitation phase. During the filter actions, the button reads: “Filter bypass.” During a precipitation phase, the button reads: “Precipitation bypass.” When pressed, button #16 instructs the instrument to stop the filter or precipitation action and start the next action.

17. Vacuum sensor pressure

The value shown in box #17 is the current pressure (in psi units) read by the vacuum sensor. If an error occurs during operation, and the user overrides the error to continue the operation, this box will not display a value.

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The computer clears all fault codes at the beginning of each new procedure. However, if you start a new procedure without correcting the cause of a fault, it will continue to re-appear.

Name

Description

Result

Correction

Paddle Temperature Sensor Failure

One of the Paddle Sensors is reporting erroneous information.

Process continues to run.

Run diagnostics. Replace Paddle Bar

assy (TDF24).

In-line Heater Temperature Sensor Failure

The device that heats the fluid before delivery is reporting a temperature that is lower than the specified range.

In-progress processes are aborted.

Run diagnostics. Replace sensor or

heater.

Paddle Heater Over Temp

One of the Paddle Sensors is reporting a temperature that is higher than the specified range.

IDF process is aborted. SDF process continues.

Run diagnostics. Replace Paddle Bar

assy (TDF24).

←

Faults

Fault Handling

When the instrument detects a problem, it generates a fault. Some faults require the instrument to abort operations. Other faults will allow the instrument to continue operation.

The screen below shows the primary sections of the instrument fault screen. The Faults screen includes sections that present information that cannot be changed by the user, along with buttons that allow

the user to modify the current operations or get additional information. The back button is available for returning to the Status screen or the Diagnostics screen depending on the function you were doing when you entered the Faults screen.

For explanation purposes, we have placed a number in each section of the screen. Your computer screen will NOT show these numbers.

1. Fault description

Box #1 contains the description of a fault that has recently occurred.

2. Reset fault after it has been repaired

Button #2 is only displayed when a low input pressure fault occurs. When button #2 is pressed, the fault is reset.

3. Next fault

Multiple faults can occur during a procedure. Pressing button #3 will walk through all of the existing faults.

List of Faults and Alerts (see www.ankom.com for the most current Fault Handing information)

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Name

Description

Result

Correction

In-line Heater Over Temp

The device that heats the fluid before delivery is reporting a temperature that is higher than the specified range.

In-progress processes are aborted.

Run diagnostics. Replace sensor or

heater.

Paddle Heater Under Temp

One of the Paddle Sensors is reporting a temperature lower than the specified range.

IDF process continues. SDF process unaffected.

Run diagnostics. Replace Paddle Bar

assy (TDF24).

In-line Heater Under Temp (IDF Amylase Phase)

The device that heats the fluid before delivery is reporting a temperature that is lower than the specified range for the Amylase phase.

IDF process continues. SDF process unaffected. Failed heater is disabled.

Run diagnostics. Replace sensor or

heater.

In-line Heater Under Temp (IDF Water Rinse)

The device that heats the fluid before delivery is reporting a temperature that is lower than the specified range for the IDF Water Rinse phase.

IDF process continues. SDF process unaffected. Failed heater is disabled.

Run diagnostics. Replace sensor or

heater.

In-line Heater Under Temp (SDF EtOH95 delivery)

The device that heats the fluid before delivery is reporting a temperature that is lower than the specified range for the SDF EtOH95 Delivery phase.

IDF process unaffected. SDF process continues. Failed heater is disabled.

Run diagnostics. Replace sensor or

heater.

In-line Heater Cooling Failure

The device that heats the fluid before delivery is reporting a temperature that is higher than the specified range for the Water Cooling phase.

In-progress processes are aborted.

Run diagnostics. Replace sensor or

heater.

E10

High Line Pressure

The pressure in one or more of the delivery tubes is higher than the specified values.

In-progress processes are aborted.

Check for plugged tube or replace output valve.

E11

Low N2 Input Pressure

The pressure at the input port is lower than the specified range.

In-progress processes are frozen in time.

The mixer is stopped. The N2 filter is disabled

until fault is corrected.

Check that N2 supply is connected and working.

E12

High N2 Input Pressure

The pressure at the input port is higher than the specified range.

Fault is reported. All in-progress processes

continue.

Adjust high pressure regulator gauge on the instrument to 50psi.

E13

Tubing Failure

At least one of the pump tubes has broken OR one of the output valves is not closing.

In-progress processes are aborted.

Replace pump tubing or repair output valve.

E14

Empty Chemical Container

The instrument detected that one or more chemical containers are empty.

A message is displayed. User can choose to retry, override, or abort. Override will prevent the error message for the rest of the run.

Fill the associated container.

E15

Supply Line Plugged

The instrument detected that one or more chemical supply lines are plugged or the associated valves are stuck in the closed position.

A message is displayed. User can choose to retry, override, or abort. Override will prevent the error message for the rest of the run.

Ensure that the supply line connector is fully engaged in the top of the container, and check for a clogged filter.

Contact ANKOM for assistance.

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←

Valve Test

Amylase Supply

(closed)

HCl Supply

(closed)

AMG Supply

(closed)

Protease Supply

(closed)

Clamp Bar A

(out)

Clamp Bar D

(out)

Clamp Bar C

(out)

Clamp Bar B

(out)

Water Supply

(closed)

EtOH95 Supply

(closed)

EtOH78 Supply

(closed)

Buffer Supply

(closed)

IDF Inlet

(open)

Waste

(open)

SDF Inlet

(open)

IDF N2

(off)

Mixer

(off)

SDF N2

(off)

Mixer

Duty Cycle

←

Diagnostics

Touch Screen Display version: A.AA SPLat controller software version: vX.XX dd-mm-yy

Pump Tube

Test

Expert Mode

(OFF)

Faults

Service

Mode

Clear IDF /

SDF lines

Digestion

Times

Heater Test

Temperatures

Pressures

Line Charge

Motor Test

Valve Test

Select a Function

AOAC 991.43

IDF/SDF

AOAC 985.29

TDF

Diagnostics

AOAC 991.43

TDF

Diagnostics Mode

To help solve problems that may occur while running the ANKOM maintenance procedures, a computer controlled Diagnostics Mode is available through the Touch Screen Display.

To access the Diagnostics Mode, press the Diagnostics button on the screen above. The following screen will be displayed.

TDF

Dietary Fiber Analyzer, and to assist with periodic

This screen has buttons for each of the available diagnostic procedures. Descriptions of each procedure are provided below.

Valve Test

When you press the Valve Test button on the Diagnostics screen, the following screen will be displayed.

When this screen is displayed, the Control Panel buttons located below the Touch Screen Display are disabled. Pressing the buttons on this screen will actuate specific valves (with the exception of the Mixer Duty Cycle button).

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The value on the Mixer Duty Cycle screen is only used during Diagnostics and in an AOAC 2009.01/2011.25 assay during the 16 hour Amylase/AMG digestion and Trizma phases. The default value for the specified AOAC 2009.01/2011.25 phases is 50.

After completing the Heater Test you should turn all heaters off.

Pressure Monitor (psi)

←

xx.x

Line 1

xx.x

Line 2

xx.x

Line 3

xx.x

Line 4

xx.x

Line 5

xx.x

Line 6

xx.x

Input Pressure

xx.x

Vacuum Sensor

←

Heater Test

Paddle Heater

Power Enable

(off)

In-line Heater

(off)

Paddle 1

(off)

Paddle 2

(off)

Paddle 3

(off)

Paddle 4

(off)

Paddle 5

(off)

Paddle 6

(off)

Heater Test

When you press the Heater Test button on the Diagnostics screen, the following screen will be displayed.

With the Paddle Heater Power Enable button set to “on”, pressing any of the Paddle 1 through Paddle 6 buttons will turn the heat on for the specific paddle. Pressing the In-line Heater button will turn the In-line Heater on and light the LED located below the Touch Screen Display on the Control Panel. The heater temperatures are shown below each heater button.

Faults

When you press the Faults button on the Diagnostics screen, the Faults screen is displayed. See the “Fault Handling” section of this manual for more information.

Pressures

When you press the Pressures button on the Diagnostics screen, the following screen will be displayed.

The readings shown on this screen for the input pressure and the lines to all six stations represent the absolute pressure. These values will dynamically change as the pressure changes.

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Running the Motor Test could leave fluid in some of the lines. After running a Motor Test, always run the Clear IDF/SDF Lines diagnostic.

←

Set Valves

Amylase Supply

(closed)

HCl Supply

(closed)

AMG Supply

(closed)

Protease Supply

(closed)

Water Supply

(closed)

EtOH95 Supply

(closed)

EtOH78 Supply

(closed)

Buffer Supply

(closed)

IDF Inlet

(open)

Waste

(open)

SDF Inlet

(open)

Output

Supply

←

Motor Test

Set Valves

Motor

Direction

(FWD)

120 RPM

240 RPM

Motor Speed

10.0

Amount (ml)

00.0

Max Vacuum (psi)

Motor Test

When you press the Motor Test button on the Diagnostics screen, the following screen will be displayed.

This diagnostic allows you to test the motor at different speeds (default is 240 RPM) and directions using different amounts of fluid. Before pressing the GO button to start the Motor Test, it is recommended that you first open at least one Supply Valve and one Output Valve. To do that, press the Set Valves button. The following screen will be displayed.

This diagnostic also displays the maximum pressure read from the vacuum sensor during the test. The value starts at zero and will display a value while the motor is running and when it stops. This can be used for troubleshooting.

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If you start a new procedure with Digestion Times that are less than the official method times, a message will be displayed on the Touch Screen Display that allows you to reset the values.

All lines should be charged before test is run. Verify that all supply containers are connected and contain the desired fluid.

Cancel

Continue

(no line charge)

Continue

(line charge)

←

Pump Tube Test

START

Status

Pressure

difference

(psi)

1 2 3 4 5

←

AOAC 991.43/985.29 Digestion Times

(Press button with number to change)

Amylase

Digestion Time

(minutes)

SDF Precipitation Time

(minutes)

Protease

Digestion Time

(minutes)

HCl/AMG

Digestion Time

(minutes)

Reset to official

method times

Save to

Permanent

Memory

Digestion Times

When you press the Digestion Times button the following screen will be displayed.

This diagnostic allows you to modify the times used by the computer for the digestion processes. This can be helpful when troubleshooting or demonstrating the instrument. If you press the Save to Permanent Memory button, the values you entered will be saved until you change them again by using this diagnostic. If you do not press the Save to Permanent Memory button, the values you entered will be saved until power is turned off, or until you change them again by using this diagnostic.

Pump Tube Test

When you press the Pump Tube Test button on the Diagnostics screen, the following screen will be displayed.

This diagnostic tests the integrity of the tubing from the pump to the inlet valves (IDF, SDF, and Waste). When you press the START button, the following screen will be displayed.

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←

Line Charge

Amylase

Protease

AMG

HCl

EtOH78

EtOH95

Water

Buffer

ALL

Temperature Monitor (°C)

←

Paddle 1

Paddle 2

Paddle 3

Paddle 4

Paddle 5

Paddle 6

In-line Heater

When you press one of the Continue buttons, the Pump Tube Test screen is once again displayed with the bottom part of the screen showing the status of each tube. This diagnostic is automatically executed at the beginning of each new IDF/SDF or TDF run.

Temperatures

When you press the Temperatures button on the Diagnostics screen, the following screen will be displayed.

The readings for the In-line Heater and all six Heating Paddles dynamically change as the temperature changes.

Line Charge

When you press the Line Charge button on the Diagnostics screen, the following screen will be displayed.

This diagnostic allows you to pump individual fluids, or all of the fluids, into the lines from the Chemical Containers to the Tubing Support Panel. If by accident you run out of any fluid during a normal IDF/SDF or TDF run, there may not be enough fluid in the lines for the next assay. In that case you should use this diagnostic to re-fill the lines that ran out.

A button will change color as the associated line is being charged. A line is charged when its associated button returns to its original green color.

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You must be careful while in Expert Mode or you could cause problems!

The IDF/SDF and TDF analyses both start filling the SDF bag with EtOH about an hour after the IDF procedure starts. Therefore, if you start an IDF procedure, and wait more than an hour to properly install your SDF bags, the instrument will deliver EtOH to the empty bottom stations, wasting EtOH, and ruining a run.

Only use this option when demonstrating or troubleshooting the instrument. If the bypass is enabled, a warning screen will be displayed at the start of a new run. At that time you can tell the computer to disable the bypass and return to the standard operating mode of waiting for the heating paddles to reach the target temperature before starting the digestion timer.

←

Service Mode

Temperature

Sensor

Corrections

IDF Count

Volume

Calibration

Bypass Amylase

Phase Initial

Heat-up

(NO)

AOAC 2009.01/

2011.25

Registration

Clear IDF/SDF Lines

When you press the Clear IDF/SDF Lines button on the Diagnostics screen, the IDF and SDF lines are cleared of any fluids. Before running this diagnostic, place small waste cups at each IDF and SDF station under the nozzles and close the Front Cover of the instrument.

Expert Mode

When Expert Mode is “ON,” all of the question screens in the IDF/SDF and TDF procedures are bypassed except for the

“Check pH manually?” and “Filter minutes OK?” screens. See the “Productivity Enhancement” section of this manual for more

information about Expert Mode.

Service Mode

When you press the Service Mode button on the Diagnostics screen, the following screen will be displayed.

Service Mode – Volume Calibration

Refer to the “QC / Calibration Checks” section of this manual for details about this diagnostic.

Service Mode – Temperature Sensor Corrections

Refer to the “QC / Calibration Checks” section of this manual for details about this diagnostic.

Service Mode – Bypass Amylase Phase Initial Heat-up

For digestions to occur at the correct temperature for the correct amount of time, the instrument’s computer waits for the heating paddles to heat up to the target temperature before starting the digestion timer. When this button displays the word “YES,” the instrument’s computer will start the digestion timer without waiting for the heating paddles to reach the target temperature.

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AOAC 2009.01/2011.25 Registration

←

xxxxx

SET

Enter registration number, then press SET.

IDF Count

←

Number of IDF runs

Service Mode – IDF Count

When you press the IDF Count button on the Service Mode screen, the following screen will be displayed.

From this screen, you can see the number of digestion operations that have been done on this instrument.

Service Mode – AOAC 2009.01/2011.25 Registration

When you purchase the upgrade to Total Integrated Dietary Fiber, ANKOM Technology will provide you with a registration number that you will use to configure your instrument to run the AOAC 2009.01 and 2011.25 methods. When you press the AOAC 2009.01/2011.25 Registration button on the Service Mode screen, the following screen will be displayed.

To enable the AOAC 2009.01/2011.25 methods, enter the 5-digit registration number provided by ANKOM Technology in the white box with the gray border and press the SET button. If you enter an incorrect number, the instrument will tell you to try again.

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Observe the spray capability during the rinse cycle of an assay.

Remove any plugged nozzles by pulling and turning them (they are friction fit to the delivery tubes).

Clean out the six holes using the Spray Tip Cleaning Tool (part # 8130) to regain unobstructed fluid passage.

Clean out any foreign matter left by the process (pressurized air may be used to blow out the holes).

Reinstall the delivery nozzles by turning and pushing them onto the delivery tubes.

Inspect the containers and container filters (at the end of the tubing inside each container) checking for any precipitate, particles, or foreign matter that may be plugging the filters or settled at the bottom of the container.

Clean the filters by running them under fast running water.

Rinse the containers out with water and let air dry as needed.

With the instrument connected to pressurized nitrogen, inspect the pressurized air lines. Listen for any audible leaks and check for cracks or kinks.

Replace tubing as needed.

Periodic Maintenance

Inspect and clean the delivery nozzles at each IDF and SDF station

On a monthly basis (or more frequently depending on usage), follow the steps below to inspect and clean the delivery nozzles.

Inspect and clean chemical containers and filters

On a monthly basis (or more frequently depending on usage), follow the steps below to inspect and clean the chemical containers and filters.

Regularly clean the instrument surfaces

Wipe down the surface of the instrument whenever spills occur or splatters are observed.

Inspect silicone tubing for wear or leaks

On a monthly basis (or more frequently depending on usage), follow the steps below to inspect the silicone tubing.

Troubleshooting & Replacement Parts

The ANKOM Technology web site has the most current troubleshooting and replacement parts information. Therefore, if you have any questions about the operation of your ANKOM visit our web site at www.ankom.com.

TDF

Dietary Fiber Analyzer, or if you need replacement parts, please

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←

Diagnostics

Touch Screen Display version: A.AA SPLat controller software version: vX.XX dd-mm-yy

Pump Tube

Test

Expert Mode

(OFF)

Faults

Service

Mode

Clear IDF /

SDF lines

Digestion

Times

Heater Test

Temperatures

Pressures

Line Charge

Motor Test

Valve Test

Select a Function

AOAC 991.43

IDF/SDF

AOAC 985.29

TDF

Diagnostics

AOAC 991.43

TDF

QC / Calibration Checks

The ANKOM instrument automatically checks its operation in multiple ways. For example, during each run the instrument automatically checks for Tube Integrity.

In addition to reviewing assay results, you can determine if the instrument is functioning properly by executing the following procedures.

Fluid Delivery

You will use the Volume Calibration diagnostic to check and calibrate the Fluid Delivery system. This diagnostic allows you to verify that the instrument is delivering the correct amount of fluid through the pump. This calibration should be done after installing new pump tubing. To run the Volume Calibration procedure, follow the steps below.

TDF

Dietary Fiber Analyzer is an automated and highly integrated instrument. Utilizing an on-board computer, the

1. Press the Diagnostics button on the screen above. The following screen will be displayed.

2. Press the Service Mode button. The following screen will be displayed.

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←

Place spare containers under IDF Delivery Nozzles to catch pre-calibration charge.

Press OK when done.

Volume Calibration

←

xx.x

Calibration Time

START

←

Service Mode

Temperature

Sensor

Corrections

IDF Count

Volume

Calibration

Bypass Amylase

Phase Initial

Heat-up

(NO)

AOAC 2009.01/

2011.25

Registration

3. Press the Volume Calibration button. The following screen will be displayed.

4. Label and weigh six small empty glass beakers.

5. Press the START button on the Volume Calibration screen above. The following screen will be displayed.

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←

Place Calibration containers under the IDF Delivery Nozzles.

Press OK when done.

Volume Calibration

6. Install extra plastic containers under all six IDF Delivery Nozzles to catch any fluid currently in the lines.

With containers in place, press the OK button. After fluid empties into the containers, the following screen will be displayed.

7. Remove the containers with fluid.

8. Install the empty pre-weighed beakers under all six IDF Delivery Nozzles and press the OK button.

While the fluids are being delivered, the following screen will be displayed.

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Volume Calibration

←

xx.x

Calibration Time

Enter average volume delivered by pressing the number input, entering the number, then pressing the “SET” button.

SET

20.0

Volume Calibration

←

Final fluid delivery in progress...

When the fluid delivery is complete, the following screen will displayed.

9. Remove the beakers with fluid and weigh them.

10. Determine the amount of fluid delivered by doing the following. Subtract the weight of the empty beakers from the

weight of the beakers that contain fluid. Given that 1g of water equals 1 ml of water, convert the weight in grams to volume in ml.

11. Press the number input box on the above screen (it shows “20.0” as the default) and enter the average ml of fluid

delivered; then press the Enter button to capture the value and exit the input screen.

12. Press the SET button. This modifies the Calibration Time and stores the new value in memory.

Temperature Sensing

The instrument uses temperature sensors on the heating paddles to monitor and control the temperature of the digestion phases. Because the sensors have a tolerance range, you can set offsets to correct for differences in temperature sensors.

To calibrate the Temperature Sensing system, follow the steps below.

1. Start an IDF/SDF analysis on the instrument without using any samples. See the “IDF/SDF Analysis” section of this

manual for more details.

2. After 15 minutes of time has elapsed in the Protease phase of the IDF process, press the Display Temperatures button

pg. 86 Rev E 12/2/14

on the Status Screen.

3. With the front cover of the instrument in the down position (as it would be during normal operation), place a standard

Thermometer in each of the six IDF bags and record on a sheet of paper the temperature readings from the Thermometer and from the sensors. The temperature should be 60°C at this time.

4. Abort the IDF/SDF run. The following screen will be displayed.

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Operator’s Manual

←

Diagnostics

Touch Screen Display version: A.AA SPLat controller software version: vX.XX dd-mm-yy

Pump Tube

Test

Expert Mode

(OFF)

Faults

Service

Mode

Clear IDF /

SDF lines

Digestion

Times

Heater Test

Temperatures

Pressures

Line Charge

Motor Test

Valve Test

Select a Function

AOAC 991.43

IDF/SDF

AOAC 985.29

TDF

Diagnostics

AOAC 991.43

TDF

Vacuum sensor:

AAAA

psi

Status

Overall Progress

Action in progress

IDF

Time remaining:

IDF Aborted

XX:XX:XX

SDF

Time remaining:

XX:XX:XX

pH required?

Display

Temps/Pressures Yes

AOAC 991.43 IDF/SDF

Main Menu

Vacuum sensor:

AAAA

psi

5. Press the Main Menu button. The following screen will be displayed.

6. Press the Diagnostics button. The following screen will be displayed.

7. Press the Service Mode button. The following screen will be displayed.

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Temperature Sensor Corrections

←

XX.X

Station 1

XX.X

x.x

Enter the difference between actual and measured temperatures.

Offsets ->

Uncorrected temperature values ->

Station 2

Station 3

Station 4

Station 5

Station 6

Save to

Permanent

Memory

←

Service Mode

Temperature

Sensor

Corrections

IDF Count

Volume

Calibration

Bypass Amylase

Phase Initial

Heat-up

(NO)

AOAC 2009.01/

2011.25

Registration

8. Press the Temperature Sensor Corrections button. The following screen will be displayed.

9. Press the white button with the gray border for each sensor that did not show the same value as the thermometer and

type in the difference between what the sensor reported and the actual temperature shown on the thermometer.

10. Press the Save to Permanent Memory button after setting the offsets. They will remain until they are changed using

this diagnostic.

11. Repeat steps 1-3 of this procedure to check that the sensors read within 2° of the Thermometer.

Heating Paddle – Mixing Operation

Computer controlled heating paddles are used along with stationary mixing pads to mix the sample with the buffer and enzyme solutions during the IDF process. To check the mixing operation of the heating paddles, follow the steps below.

1. Start an IDF/SDF analysis on the instrument without using any samples. See the “IDF/SDF Analysis” section of this

manual for more details. When Clamp Bar A closes (at the beginning of the Amylase phase), the heating paddles will start actuating.

2. Count the number of heating paddle actuations that occur in 15 seconds.

You should see 10-15 actuations within 15 seconds.

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Enzyme Dilution Table

Enzyme

Dilution

α-Amylase (ANKOM Concentrate)

Dilute 5 ml of amylase to 25 ml

Protease (ANKOM Concentrate)

Dilute 5 ml of protease to 25 ml

Amyloglucosidase (ANKOM Concentrate)

Dilute 5 ml of Amyloglucosidase to 25 ml

The ANKOM

TDF

Dietary Fiber Analyzer delivers 1 ml of each enzyme solution to each of the six stations per run (6 ml of each enzyme solution). If you are NOT using the ANKOM Concentrate enzymes, you must prepare enzyme solutions that include 0.02% w/v of Sodium Azide (to prevent microbial growth) and the following enzyme concentrations:

α-Amylase:

150 Ceralpha Units per ml

Protease:

35 Tyrosine Units per ml

Amyloglucosidase:

652 Units per ml

To achieve the appropriate volume in the filter bags, the concentrations of the HCl and NaOH solutions were modified to accomplish the pH adjustment in a manner equivalent to the AOAC 985.29 method.

To avoid microbial growth, make all buffer solutions fresh and do NOT allow them to sit unused for more than two days.

Appendix A – Reagents (AOAC 991.43, AOAC 985.29)

Use Distilled Water throughout.

Solutions common to both AOAC 991.43 and AOAC 985.29 methods

(a) Ethanol 95%. (b) Ethanol 78%—Place 821 ml of 95% ethanol into a 1 L volumetric flask. Dilute to volume with H2O. (c) Enzyme solutions—To make your enzyme solutions for use in the ANKOM

ANKOM Concentrate enzymes (TDF80/TDF81, TDF82/TDF83, TDF84/TDF85) with H2O according to the Enzyme Dilution Table below.

TDF

Dietary Fiber Analyzer, dilute

(d) Diatomaceous earth (DE)—(ANKOM XTC, or equivalent). (e) Acetone—reagent grade.

Solutions unique to the AOAC 991.43 method

(f) MES—2-(N-Morpholino)ethanesulfonic acid (No. M-8250, Sigma Chemical Co., or equivalent). (g) TRIS—Tris(hydroxymethyl)aminomethane (No. T-1503, Sigma Chemical Co., or equivalent). (h) MES-TRIS buffer solution—0.05M MES, 0.05M TRIS, pH 8.2 at 24°C. Dissolve 19.52 g MES and 12.2 g TRIS in 1.7

L H2O. Adjust pH to 8.2 with 6N NaOH, and dilute to 2 L with H2O. (Note: It is important to adjust pH to 8.2 at 24°C. However, if buffer temperature is 20°C, adjust pH to 8.3; if temperature is 28°C, adjust pH to 8.1. For deviations between 20 and 28°C, adjust by interpolation.)

(i) Hydrochloric acid solution (AOAC 991.43)—0.561N. Dilute 93.5 ml of 6N HCl to 1 L with H2O.

Solutions unique to the AOAC 985.29 method

(j) Hydrochloric acid solution (AOAC 985.29)—0.65N. Dilute 325 ml of 1N HCl to 500 ml with H2O. (k) Phosphate buffer solution—0.08M, pH 6.0. Dissolve 1.40 g Sodium Phosphate dibasic, anhydrous (Na2HPO4) (or 1.75

g dihydrate) and 9.68 g Sodium Phosphate monobasic monohydrate (NaH2PO4) (or 10.94 g dihydrate) in a 1 L volumetric flask. Dilute to 1 L with H2O. Adjust pH to 6.0.

(l) Sodium Hydroxide Solution—0.55N. Dissolve 11.00 g Sodium Hydroxide (NaOH ACS) in a 500 ml volumetric flask.

Dilute to 500 ml with H2O.

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These instructions are available with additional pictures in Service Procedure 208 located at www.ankom.com.

←

Line Charge

Amylase

Protease

AMG

HCl

EtOH78

EtOH95

Water

Buffer

ALL

Connect to HCl tube

Connect to right-side tubes

Connect to left-side (enzyme) tubes

Connect to 78% EtOH container

Appendix B – Instrument Out-of-use Procedure

The fluid lines can become obstructed when the instrument is left unused for one week or longer. To avoid fluid line obstruction, execute the following procedure BEFORE the instrument is left unused for one week or longer.

1. Flush and purge the lines.

1.1 With the instrument turned on, press the Diagnostics button on the Touch Screen Display.

1.2 Press the Line Charge button on the Touch Screen Display. The following screen will be displayed.

1.3 Fill the Enzyme Containers on the left side of the instrument with Distilled Water. The enzyme lines are first flushed

with Distilled Water to avoid any precipitation from occurring due to an interaction between the enzymes and the EtOH.

1.4 Press the Amylase button on the Touch Screen Display. This will draw water into the Amylase line. Do this a

second time to fully flush the line.

1.5 To flush the Protease and AMG lines, repeat step 1.4 using the Protease and AMG buttons on the Touch Screen

Display.

1.6 Disconnect Enzyme, HCl, EtOH78, EtOH95, Water, and Buffer containers. Store or dispose of all fluids properly.

1.7 Obtain the TDF70 Line Flush Tubing Assembly shown below. Note the nine connection points on the Tubing

Assembly.

1.8 Attach the Line Flush Tubing Assembly to the instrument and to the 78% EtOH Container as shown above.

1.9 Fill the 78% EtOH Container with 220ml of 78% EtOH.

1.10 Press the ALL button on the Touch Screen Display. This will draw 78% EtOH into all of the lines. Do this a second

time to fully flush the lines. This process will drain the 78% EtOH Container.

1.11 With the 78% EtOH Container empty, press the ALL button on the Touch Screen Display to purge the fluid from the

lines by sucking air into them. Press the ALL button a second time to fully clear the lines. There should now be minimal fluid remaining in the lines. If a significant amount of fluid is observed, press the ALL button again until the lines are clear (some droplets will remain). During the last line charge you should see no fluid being pumped out to waste.

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2.1 Unscrew the six acorn nuts on the

back of the instrument and remove the clear back panel to access the pump area.

2.2 Un-latch the two pump doors so

they are open.

2.3 Reinstall the clear back panel and

tighten the acorn nuts.

When returning the instrument to service after a period of non-use, you must close and latch the pump doors and charge all of the lines before running. See the IDF/SDF Analysis, TDF Analysis, or Diagnostics sections of this manual for information about charging the lines.

Acorn Nuts (the top two are behind the black torsion bar)

Cable Tie

Pump Doors

2. Open the pump doors.

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The XT4 Filter Bag is used in the ANKOM XT Fat Extractors. The bag has a porosity of 1-4 microns. As a result, the likelihood of sample loss is eliminated.

Number all of the XT4 Filter Bags to be used for testing.

Record the weight of each XT4 Filter Bag.

It is important to note that the weight of the XT4 Filter Bag must be included in the analysis calculations in the following ways:

 For an IDF/SDF analysis, add the weight of the XT4 bag to the weight of

the IDF bag.

 For a TDF analysis, add the weight of the XT4 bag to the weight of the

SDF bag.

Record the weight of each sample to be de-fatted in the appropriate XT4 Filter Bags.

Heat seal the Filter Bags.

With the heat sealer set at 5-6, seal the Filter Bag within 4mm of its open end. Be sure to hold the sealer arm down for 2-3 seconds after the red sealer light turns off to properly cool the seal.

Appendix C – De-fatting Procedure

According to the AOAC 985.29E method (and referred to in the AOAC 991.43 method), any sample with a fat content ˃10% must have the fat removed before determining the amount of Dietary Fiber.

In the conventional method, most labs perform the de-fatting step in the same beaker in which the enzyme digestion occurs. While this method would work with the ANKOM beaker into the IDF or IDF Flow-Thru bags less than efficient. As a result, we have developed a de-fatting method using an ANKOM XT4 Filter Bag in an effort to eliminate sample loss.

To de-fat your sample prior to running the IDF/SDF or TDF analyses, follow the steps below.

TDF

Dietary Fiber Analyzer, many would find transferring the sample from the

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Soak the bags in Petroleum Ether.

5.1 Place all XT4 Filter Bags into a beaker or other container.

5.2 Pour enough Petroleum Ether in the container to cover the bags and allow them to

soak in the solvent for 10 minutes.

Remove the Filter Bags from the container and allow the solvent to evaporate until the bags are completely dry.

Carefully cut the XT4 Filter Bags inside the IDF Filter Bags or the IDF Flow-Thru Bags on the TDF instrument.

To get the XT4 Filter Bag pieces into the TDF bags, hold each XT4 Filter Bag inside the appropriate TDF bag and cut the XT4 into approximately a dozen pieces being careful not to lose any sample. If any sample sticks to the scissors, brush it off into the TDF bag.

For a TDF analysis, make sure all XT4 pieces drop from the IDF Flow- Thru Bags into the SDF Filter Bags.

Use tongs if necessary to help move the pieces.

When performing the Ashing procedure, use an XT4 ash BLANK value in the calculation.

10.

When performing the Kjeldahl procedure, in order to appropriately digest the extra XT4 bag material, add 4-5ml more sulfuric acid than is specified in the "Protein Determination" section of this Operator's Manual.

The XT4 Filter Bags do NOT contain any protein.

A small amount of PTFE polymer will NOT digest during the Kjeldahl procedure. This is of NO concern.

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TDF51 – Rinse Stand assy

XTC – Diatomaceous Earth (DE)

1915 – Heat Sealer (120v) 1920 – Heat Sealer (220v)

DF-S – SDF Filter Bags (50)

DF-I – IDF Filter Bags (50)

DF-FT – Flow-thru Filter Bags (50)

Appendix D – Accessories (sold separately)

Contact ANKOM Technology or an authorized dealer to order accessories.

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Automation saves time and money!

ANKOM Technology is an international company with products that include…

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 Crude Fiber (AOCS Ba 6a-05), ADF, NDF  Automatically adds solutions and rinses

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 Official Method AOCS Am 5-04  Fully automatic  Solvent recovery at 97% or greater

 Batch process - up to 15 samples at one time

RF Gas Production System

 High sensitivity pressure measurement  Applications include: Biomass-to-Energy analysis (e.g., Ethanol, methane, etc.),

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 Wireless Computer control and data storage

Chemicals

 A wide variety of chemicals used for many different lab operations

 Pre-mixed solutions available

Please visit our web site at www.ankom.com for more information.

2052 O’Neil Rd, Macedon NY 14502 Telephone: (315) 986-8090 Fax: (315) 986-8091 www.ankom.com