Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume
46, Number 3, March 2008, pp. 225-232
Simultaneous Determination of Four Water-Soluble
Vitamins in Fortified Infant Foods by Ultra-Performance Liquid
Chromatography Coupled with Triple Quadrupole Mass Spectrometry
Baiyi Lu[1], Yiping Ren[2], Baifen Huang[2],
Wenqun Liao[2], Zengxuan Cai[2], and Xiaowei Tie[2]
[1]Department of Food Science and Nutrition, College of Biosystem
Engineering and Food Science, Zhejiang University, Hangzhou 310029,
China and
[2]Zhejiang Provincial Center for Disease Prevention
and Control, Hangzhou 310007, China
A novel ultra-performance liquid chromatography
electrospray ionization tandem triple quadrupole mass spectrometry
method for the simultaneous determination of four water-soluble
vitamins, including vitamin B5 (VB5), vitamin B8 (VB8), vitamin
B9 (VB9), and vitamin B12 (VB12) in fortified infant foods is
developed and validated. A reverse phase UPLC separation system
consisting of a Waters ACQUITY UPLC BEH C-18 column (2.1 mm × 100
mm i.d., 1.7 µm) and a binary gradient acetonitrile–water
mobile phase is applied for the separation of the four water-soluble
vitamins. Formic acid is spiked into the mobile phase to enhance
the ionization efficiency. Tandem MS–MS analysis is performed
in multi-reaction monitoring mode (MRM). Product-ion traces at
m/z 220.1 → 89.9 for VB5, 245.1 → 227.1 for VB8, 442.3 → 295.2
for VB9, and 678.9 → 147.0 for VB12 are used for quantitation
of the corresponding vitamins, and traces at m/z 455.5 → 308.0
are used for methotrexate (internal standard). Limits of quantitation
(LOQs) are 0.016, 0.090, 0.020, and 0.019 µg/L for VB5,
VB8, VB9, and VB12, respectively. Intra- and inter-day precisions
for the determination of the four vitamins are better than 6.84%
and 12.26% in relative standard deviations, and recoveries for
the four vitamins are in the range of 86.0~101.5%. The developed
approach is applied for the determination of the trace amounts
of the vitamins in fortified milk powers and fortified rice powers.
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