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
a.) Pipette and appropriate amount of each folate stock solution (EXCPET THE INTERNAL STANDARD STOCK SOLUTION AND PTEROLYTRI-y-GLUTAMIC ACID) into a 10mL low-actinic volumetric flask. b.) Three separate solutions (one mixture and two individual solutions) are prepared as detailed in the following table. c.) 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. d.) Bring to volume with Folate Intermediate Standard Solvent and store approximately -20 o C. Stable for up to 30days.
Table 3: Preparation intermediate standard solutions
Mixture Intermed or Individual std. Soln
Stock std. Concn (mcg/mL)
Stock std. Volume (uL)
Intermed std Soln #
Folate compd #
Final Volume* (mL)
Concen (mcg/mL)
Stock Standard
Shircks (6S)-5-Formyl-5,6,7,8- tetrahydrofolic acid
1
607
356
21.6092
2
10-Formylfolic acid
597
335
19.9995
Mixture Intermed Std. Soln
4
10-methylfolic acid
529
378
19.9962
I
10
Shircks (6R,S)-5,10-methenyl- 5,6,7,8-tetrahydrofolic acid
5
587
368
21.6016
Shircks (6S)-5,6,7,8- tetrahydrofolic acid
7
542
369
19.9998
Indiv. Intermed. Std soln Indiv. Intermed. Std soln Conjugase check soln
Shircks (6R,S)-5-methyl-5,6,7,8- tetrahydrofolic acid
II
3
584
371
10
21.6664
III
8
Shircks Folic acid
555
360
10
19.98
Conjugase check soln
6
pteroyltri-y-L-glutamic acid
911
222
10
20.2242
*Make up final volume to 10 mL with Folate Intermediate std. solvent.
7.) Internal Standard Intermediate Standard Solution (20ug/mL):
8
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