ANKOM TDF Dietary User Manual

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

Rev E 12/2/14 pg. 5

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

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

Rev E 12/2/14 pg. 7

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

pg. 8 Rev E 12/2/14

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

Rev E 12/2/14 pg. 9

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

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:

Rev E 12/2/14 pg. 11

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

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

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

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

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

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.

Rev E 12/2/14 pg. 17

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. 18 Rev E 12/2/14

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.

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

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

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

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

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

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

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

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

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

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

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ANKOM TDF Dietary User Manual

Specifications and Main Features

Frequently Asked Questions

User Manual