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Silliker
Chem, Res. Center Crete, IL – Validation of a LC-MS/MS Method for Vitamin K Analysis
Silliker Laboratories, Chemistry Research Center, Date: 1/30/15
4
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
VitK-01 w/SLV
FOR ERP USE ONLY
DO NOT DISTRIBUTE