Chen Pei, Wolf Wayne R
United States Department of Agricultural Research Service, Beltsville Human Nutrition Research Center, Food Composition Laboratory, Beltsville, MD, 20705, USA.
Anal Bioanal Chem. 2007 Apr;387(7):2441-8. doi: 10.1007/s00216-006-0615-y. Epub 2006 Nov 8.
The purpose of this study was to optimize chromatographic and detection conditions for the simultaneous determination of water-soluble vitamins in multi-vitamin dietary supplements using a single chromatographic run. An approach using liquid chromatography with diode array and/or mass spectrometry for quantitation of seven B-complex vitamins [thiamine (B(1)), riboflavin (B(2)), nicotinamide (B(3)), pyridoxine (B(6)) pyridoxine, biotin, pantothenic acid, and folic acid] in multi-vitamin/multi-mineral daily supplements is described. This approach utilizes a reversed phase C18 column (4 mum; i.d.: 250x2.0 mm) with a gradient mobile elution profile, performed at a flow rate of 0.25 ml/min. After a 5-min isocratic elution at 100% A (0.1% formic acid in water), a linear gradient to 50% A and 50% B (0.1% formic acid in acetonitrile) at 15 min and then to 5% A and 95% B at 17 min was employed. Detection was performed with a photodiode array detector (DAD) in sequence with a triple-quad mass spectrometer in the multiple reaction mode (MS-MRM). Although good chromatographic separation of ascorbic acid was also obtained in extracts from multi-vitamin/multi-mineral supplements, the ascorbic acid could not be quantified properly due to rapid oxidation catalyzed by the minerals. This method was initially applied to determine water-soluble vitamins in representative multi-vitamin/multi-mineral tablets following the extraction of ground samples with a phosphate buffer (10 mM, pH 2.5). For multi-vitamin supplement tablets, this approach does not require any sample clean-up/pre-concentration steps except for centrifugation and filtration of the extract.
本研究的目的是优化色谱和检测条件，以便在一次色谱运行中同时测定多种维生素膳食补充剂中的水溶性维生素。本文描述了一种使用液相色谱结合二极管阵列和/或质谱法定量多种维生素/多种矿物质日常补充剂中七种B族复合维生素[硫胺素（B1）、核黄素（B2）、烟酰胺（B3）、吡哆醇（B6）、生物素、泛酸和叶酸]的方法。该方法采用反相C18柱（4μm；内径：250×2.0mm），梯度流动洗脱，流速为0.25ml/min。在100%A（0.1%甲酸水溶液）下等度洗脱5分钟后，在15分钟内线性梯度至50%A和50%B（0.1%甲酸乙腈溶液），然后在17分钟内至5%A和95%B。检测采用光电二极管阵列检测器（DAD），并依次与三重四极杆质谱仪在多反应监测模式（MS-MRM）下进行。尽管在多种维生素/多种矿物质补充剂的提取物中也获得了抗坏血酸的良好色谱分离，但由于矿物质催化的快速氧化，抗坏血酸无法正确定量。该方法最初应用于测定具有代表性的多种维生素/多种矿物质片剂中的水溶性维生素，方法是用磷酸盐缓冲液（10mM，pH2.5）提取研磨后的样品。对于多种维生素补充剂片剂，除了对提取物进行离心和过滤外，该方法不需要任何样品净化/预浓缩步骤。
