Waters Biopharmaceutical Training Kit - Manual (Page 2)

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[ CARE AND USE MANUAL ]
Biopharmaceutical Training Kit
II. STANDARD P REPARATION
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 H
2
O: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.
4. Vortex and sonicate for 10 minutes.
5. Label as “75:25 H
2
O:ACN + 0.1% FA”.
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.
5. Label as “50:50 H
2
O:ACN + 0.1% FA”.
Table 2. Additional Components Required (not included in kit)
Solvents Containers Pipettes (and tips)
LCMS-grade acetonitrile
5 x 1.5 mL Waters disposable
glass vials (with caps)
Recommended Part # 186000327c
20 µL pipette
LCMS-grade or Milli-Q ® water
2 x 20 mL scintillation vials with
teflon-lined screw caps or 50 mL
polypropylene falcon tubes for
making up and storing diluents
200 µL pipette
3% hydrogen peroxide, approximately
2 mL (quality that is available and can
be purchased at local drug or grocery
store will suffice)
2 x 100 mL graduated
measuring cylinders
1,000 µL pipette and 5,000 µL pipette
Caution: Use solvents of the appropriate
grade. Waters recommends Fisher Scientific
LC/MS grade high purity solvents. Caution: All glassware must be clean,
rinsed with LCMS-grade methanol and
dry prior to use. Do not use any detergents. 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.