AOAC SPIFAN ERP Meeting Book-March 16, 2016

VitK-01 w/SLV FOR ERP USE ONLY DO NOT DISTRIBUTE

Silliker Chem, Res. Center Crete, IL – Validation of a LC-MS/MS Method for Vitamin K Analysis

Method:

Vitamin K Analysis in infant formulas and adult nutritionals

Definition: The described method is an Ultra pressure LC-MS/MS method for determining Vitamin K1 (cis and trans), vitamin K2-7 and vitamin K2 -4 infant formulas and adult nutritional matrices after enzyme digestion.

Analyte Scope:

Vitamin K1 cis and trans Vitramin K2-7 Vitramin K2-4

Matrix Scope: This method is applicable to infant formula and adult nutritional samples. Principle: An aliquot of the sample is mixed with internal standard. Fat in the sample is hydrolyzed by lipase. Vitamin K in the sample after lipase treatment is extracted in a solvent. The reconstituted extract is analyzed by LC-MS/MS for vitamin K.

Range:

1-12.5 µg/100.

Applicable Matrices:

Infant formula and adult Nutritional Formulas

Equipment:

Shimadzu UFLC system connected to API 3200 triple quad mass spec detector API 3200 mass spec detector Column: YMC CAROTENOID S-3 2.0X150mm Part# CT99S03-1502WT Turbovap solvent Evaporator Centrifuge Shaker Vortex Mixer Water bath (37 o C)

Chemicals:

a.) Lipase – Sigma #L1754-100G b.) Potassium phosphate monobasic c.) 10N NaOH: 10N NaOH obtained from Fischer (Fisher #746616) d.) K1 IS – Aldrich part#705470; 2-methyl-3-phytyl-1,4-naphthoquinone; Vitamin K1-d7 (5,6,7,8-d4, 2-methyl-d4)

e.) Vitamin K1 Standard – Chromadex #ASB-00022816-001 f.) Vitamin K2-4 Standard – Chromadex #ASB-00022821-025 g.) Vitamin K2-7 Standard – Chromadex #ASB-00022822-005

h.) Iso-Octane – Fisher #O2964 i.) Ethanol – Sigma #459836-2L

Silliker Laboratories, Chemistry Research Center, Date: 1/30/15

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