AOAC SPIFAN ERP Meeting Book-March 16, 2016

2011.06 (Fol-22) w/SLV FOR ERP USE ONLY DO NOT DISTRIBUTE

Silliker Chem, Res. Center Crete, IL – Report of A Validation of LC-MS/MS Method for Folate Analysis

Table 4: Preparation of Calibration Standard Solutions:

Internal Std Soln. (2 µ g/mL) (µL)

Neutr Elution Solv. ml

Working Internal Std Soln. (40ng/mL) (µL)

Conc. #3 (ng/mL )

Conc. #1 (ng/mL )

Conc. #7 (ng/mL )

Conc. #4 (ng/mL )

Conc. #2 (ng/m L)

Intermed iate STD Soln.

Final Vol (mL)

Conc. #8 (ng/ mL)

Conc. #5 (ng/mL)

K STD

25 uL Intermed . Std. soln. # I, II, III 10 uL Intermed . Std. soln. # I, II, III 0.1 mL Work std Soln A 0.1 mL Work std Soln B 0.1 mL Work std Soln C 0.1 mL Work std Soln D 0.05 mL Work std Soln D

20

905

1.0 mL

499.99 5

499.90 5

499.9 875

A

0

499.5

541.66

540.23

540.04

20

950

1.0 mL

216.66 4

216.09 2

199.99 8

199.96 2

199.9 95

B

0

199.8

216.016

0

0

1.0 mL

49.999 5

49.990 5

49.99 875

C

900

49.95

54.166

54.023

54.004

0

0

1.0 mL

21.666 4

21.609 2

19.999 8

19.996 2

19.99 95

D

900

19.98

21.6016

0

0

1.0 mL

4.9999 5

4.9990 5

4.999 875

E

900

4.995

5.4166

5.4023

5.4004

0

0

1.0 mL

2.1666 4

2.1609 2

1.9999 8

1.9996 2

1.999 95

F

900

1.998

2.16016

0

0

1.0 mL

1.0833 2

1.0804 6

0.9999 9

0.9998 1

0.999 975

G

950

0.999

1.08008

11.) Folate Rat Plasma Test Solution (20ug/mL Pteroyltri-y-glutamic acid (FA-glu 3 )): a.) Pipette and appropriate amount of FA-glu 3 stock solution into a 10mL low-actinic volumetric flask. b.) Add approximately 5mL of Folate Intermediate Standard Solution Solvent and sonicate in water bath set at 40 o C for about 10min to thoroughly mix. c.) Bring to volume with Folate Intermediate Standard Solvent and store approximately -70 o C. Stable for up to 180days. Dissolve 0.1g protease in 50 mL of water. Filter through glass wool if necessary and store at 4-8 °C. until use. Prepare fresh on day of use. Each sample requires 1mL. 12.) Protease Solution (2 mg/mL):

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