AOAC OMA# 2011.06 Final Action Review-OMB

2011.06 (September 2017)-Fol-22 MLT Report

FOR ERP USE ONLY DO NOT DISTRIBUTE

6. SPE Columns – mixed mode weak anion exchange (WAX SPE) Phenomenex no. 8B-S038UBJ 7. LC Column: Waters ACQUITY UPLC HSS T3 Column (100Å, 1.8 µm, 2.1 mm X 50 mm), part# 186003538 or equivalent. 8. Refrigerator (capable of cooling to 4 o C) 9. Freezer (capable of cooling to -20 o C) 10. Water Bath, with shaker 11. Centrifuge (capable of up to 1400 x g) equivalent Sorvall Legend X1R and rotor F15-8x50c with radius (Rmax 10.4 cm). . 12. Microfuge (capable of up to 9500 xg) 13. Adjustable (100 – 1000 µl) mechanical pipette 14. Vortex mixer

E. Supplies:

1.) Disposable Glass Culture Tubes

Fisher Cat. No. 14-961-30

16x125mm 2.) 50mL centrifuge tubes

Fisher Cat. No. 14-375-150

3.)0.45 micron PVDF syringe filters 4.) Microcentrifuge tubes; 2.0 mL Fisher cat. No. 02-681-344 5.) SPE cartridge: Phenomenex Part# 8B-S038UBJ. (A mix mode weak anion exchange and reverse phase solid-phase extraction cartridge). Order for batch# 70 reserved for the study from Phenomenex in US. (Phone 310.212.0555 Ext. 2348, contact: Akeema Onalaja. 6.) Flasks, Volumetric: assorted sizes Pipettes, disposable glass Pasteur pipettes

7.) Pipettes, volumetric (Class A): assorted sizes 8.) Repeating pipette : 5 mL and 25 mL or equivalent 9.) Beakers; assorted sizes

F. Chemicals/Standards:

Most of the chemicals used are LCMS grade unless specified. Product numbers & suppliers, when listed, reflect those used in validation

I - Chemicals used as Standard and Internal Standard 1.) Folic Acid (Standard)

Schircks Laboratories

Cat. No. 16.203

2.) (6R,S)-5-Methyl-5,6,7,8-(6S)-5,6,7,8- Tetrahydrofolic acid calcium salt Schircks Laboratories

Cat. No. 16.235

3.) (6S)-5-Formyl-5,6,7,8- Tetrahydrofolic acid, calcium salt Schircks Laboratories

Cat. No. 16.221

Collaborative Study Report: Method AOAC 2011.06 (Folate by LC-MS/MS) 25