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

Adult Nutritional Powder Milk Protein Based

2

11750017V3

POWDER

9

4.42

Infant Formula Powder Partially Hydrolyzed Milk Based Infant Formula Powder Partially Hydrolyzed Soy Based

3

1172572116

POWDER

9

13.54

4

117257651Z

POWDER

9

12.56

5

Adult Nutritional Powder Low Fat

00394RF00

POWDER

9

4.38

6

Child Formual Powder

00412RF00

POWDER

9

5.91

7 8 9

Infant Elemental Powder

00403RF00 D04HTCVV

POWDER POWDER POWDER

9 9 9

2.92 6.35 6.56

Infant Formula Powder Milk Based Infant Formula Powder Soy Based

E29JVLV

SPIFAN CONTROL MILK FORM

10

Infant Formula RTF Milk Based

LIQUID

9

6.65

11 12

Adult Nutritional RTF High Protein Adult Nutritional RTF High Fat

00414RF00 00406RF00

LIQUID LIQUID

9 9

3.68 2.82

* Minor Includes = 5-CHO-THF + THF + 10-CH3-FA + 10-CHO-FA + Methenyl-THF

Conclusions:

1. A LC-MS/MS method was developed and validated for analysis of different folate vitamers. 2. HPLC conditions and mass spec parameters were optimized. 3. The composition of extraction buffer and analysis buffer were adjusted to provide best analytical response of the analytes. 4. SPE conditions and elution solvents were selected to provide best recovery of the analytes. 5. The method accuracy was evaluated and established by analysis of various reference materials. 6. The method employed to analyzed a series of the SPIFA samples. The powder samples were reconstituted to the liquid form and RTF samples were analyzed as is. The results expresse as mcg/100g of total folate as folic acid (sum of the detected folate compounds after calculation as folic acid) 7. The method precision was evaluated for folate analysis in the SPIFAN samples and was found satisfactory. 8. Results of selected folate vitamers are presented for certain reference material. 9. Five SPIFAN samples (two powders and three RTF) were spiked with targeted folate compounds at three different levels (spike Level I, II and III). % spike recovery of folates as folic acids ranged from 92 to 104%. All the spiked samples provided spike recovery within the recovery range 90-110% stipulated by the SMPR. Folate recovery analysis was satisfactory in all of the studied SPIFAN matrices.

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