Waters Biopharmaceutical Training Kit User Manual
[ CARE AND USE MANUAL ]
Biopharmaceutical Training Kit
CONTENTS
I. INTRODUCTION
II. STANDARD PREPARATION
III. ADDITIONAL RESOURCES
IV. ORDERING INFORMATION
The Biopharmaceutical Training Kit comes in two separate boxes. The Biopharmaceutical Training Standard Kit (see Table 1) contains
the standards. This kit is shipped at ambient temperatures, however upon receipt of this kit, please store at -20 °C to maximize long
term stability. The Biopharmaceutical Training Prep Kit (see Table 2) contains the materials needed for solubilization and standard prep.
This kit is shipped at ambient temperatures and can be stored at room temperature. Once any component is opened, use right away for
optimum results.
I. INTRODUCTION
The Biopharmaceutical Training Kit is designed to provide trainers
and customers with high quality standards that can be utilized as a
starting point for running biopharmaceutical applications on their
Waters UPLC inlet with Tof or Q-Tof mass spectrometry detection
systems. The training kit pulls together many of the individual
standard samples that are often used in demos and training classes,
including reference compounds for calibration and lockmass. T his
kit provides a foundation for biopharmaceutical LC-MS analysis and
can be used for optimization of methods prior to running valuable
samples to ensure system performance and confidence in results.
Table 1. Kit Components for Biopharmaceutical Training Standards (P/N 186006921)
Part # Contents Description of Components Acid
186006921-1 1 x 15 mL Horse Heart Myoglobin 2.5 mg 15 mL Clear Nalgene Bottle
1860 0 6921-2 1 vial Intact mAb Mass Check Standard 1 mg
1860 0 6921-3 1 vial Glu-fibrinopeptide B 0.5 mg 4 mL Amber Screw Top Vial
1860 0 6921-4 1 vial MassPREP Enolase Digest 1 nmol
1860 0 6921-5 1 vial Reverse Peptide Ser-Asp-Gly-Arg-Gly 1.0 mg of each 4 mL glass amber bottle with screw top
1860 0 6921-6 1 vial Reverse Peptide Gly-Arg-Gly-Asp-Ser 1.0 mg of each 4 mL glass amber bottle with screw top
Amount
per Vial
Container
TruView™ Max Recovery with non-slit PTFE/
silicone Septa (p/n 186005668CV)
TruView Max Recovery with non-slit PTFE/
silicone Septa (p/n 186005668CV)
Table 2. Kit Components for Biopharmaceutical Prep Kit (P/N 186007517)
Part # Contents Description of Components Acid
186007517-1 1 x 2 mL Formic Acid 2 mL 2 mL Ampoule
186007517-2 1 x 30 mL Sodium Cesium Iodide 2 μg/μL 30 mL Nalgene
290002010 3 30 mL Nalgene Fludics Bottle (with cap) Empty 30 mL Nalgene Fludics Bottle (with cap)
Amount
per Vial
Container
[ CARE AND USE MANUAL ]
Table 2. Additional Components Required (not included in kit)
Solvents Containers Pipettes (and tips)
5 x 1.5 mL Waters disposable
LCMS-grade acetonitrile
®
LCMS-grade or Milli-Q
3% hydrogen peroxide, approximately
2 mL (quality that is available and can
be purchased at local drug or grocery
store will suffice)
Caution: Use solvents of the appropriate
grade. Waters recommends Fisher Scientific
LC/MS grade high purity solvents.
water
glass vials (with caps)
Recommended Part # 186000327c
2 x 20 mL scintillation vials with
teflon-lined screw caps or 50 mL
polypropylene falcon tubes for
making up and storing diluents
2 x 100 mL graduated
measuring cylinders
Caution: All glassware must be clean,
rinsed with LCMS-grade methanol and
dry prior to use. Do not use any detergents.
20 µL pipette
200 µL pipette
1,000 µL pipette and 5,000 µL pipette
Caution: All pipettes used to make up diluents
or samples must be calibrated and within
specification. Use appropriate pipettes and
tips during the sample dilution steps.
II. STANDARD PREPARATION
Section 1. Diluent Preparation
a. 90:10 water:acetonitrile + 0.1% formic acid
1. Using a 5,000 µL pipette, add 8.990 mL of LC/MS grade
or Milli-Q water to a clean vessel such as a 20 mL glass
scintillation vial or a 50 mL polypropylene Falcon tube.
2. Using a 1,000 µL pipette, add 1.000 mL of LC/MS grade
acetonitrile. Vortex.
3. Using a 20 µL pipette, add 10.00 µL of formic acid.
4. Vortex.
5. Label as “90:10 H2O:ACN + 0.1% FA”.
b. 75:25 water:acetonitrile + 0.1% formic acid
1. To a clean, dry 100 mL graduated cylinder, fill to the
25.00 mL mark with LC/MS grade acetonitrile.
2. Add LC/MS grade or Milli-Q water to the 100.00 mark.
3. Using a 200 µL pipette, add 100 µL of formic acid.
c. 0.2% formic acid
1. Using a 5,000 µL pipette, add 9.980 mL LC/MS or Milli-Q
water to a clean vessel such as a 20 mL glass scintillation vial
or a 50 mL polypropylene Falcon tube.
2. Using a 20 µL pipette, add 20.00 µL of formic acid.
3. Vortex.
4. Label as “0.2% FA”.
d. 50:50 water:acetonitrile + 0.1% formic acid
1. To a 100 mL graduated cylinder, add acetonitrile to the
50.00 mL mark.
2. Add LC/MS or Milli-Q grade water to fill to the 100.00 mark.
3. Using a 200 µL pipette add 100 µL formic acid into the
graduated cylinder.
4. Sonicate for 5 minutes.
4. Vortex and sonicate for 10 minutes.
5. Label as “75:25 H2O:ACN + 0.1% FA”.
5. Label as “50:50 H
O:ACN + 0.1% FA”.
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Biopharmaceutical Training Kit
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