Comparison of different extraction methods for simultaneous determination of B complex vitamins in nutritional yeast using LC/MS-TOF and stable isotope dilution assay
Comparison of different extraction methods for simultaneous determination of B complex vitamins in nutritional yeast using LC/MS-TOF and stable isotope dilution assay
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DOI：
10.1007/s00216-012-6538-x
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发表时间：
2013-02-01
影响因子：
4.3
通讯作者：
Nisamedtinov, Ildar
中科院分区：
文献类型：
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作者：
Haelvin, Kristel;Paalme, Toomas;Nisamedtinov, Ildar
研究了LC/MS-TOF法结合稳定同位素稀释法测定食品中维生素B1、B2、B3、B4、B5、B6的含量。营养酵母粉被用作模型食品基质。在甲酸铵缓冲液中将酸提取与各种酶处理进行比较，以找到将更复杂的维生素转化为与所用同位素标记的内标物相同形式的合适方法。酶制剂α-淀粉酶、takadastase、β-葡萄糖苷酶和酸性磷酸酶都能够释放硫胺素和核黄素。糖化酶制剂α-淀粉酶和takadiastase也表达蛋白水解的副作用，导致小肽的形成，干扰硫胺素和核黄素的质谱。烟酰胺和泛酸分别从NAD（+）和CoA中释放，不能用任何研究的酶制剂实现。盐酸萃取在121 A度C为30分钟，被认为是破坏泛酸，但增加了吡哆醛的释放。
The application of LC/MS-TOF method combined with stable isotope dilution assay was studied for determination of thiamine, riboflavin, nicotinamide, nicotinic acid, pantothenic acid, pyridoxal, and pyridoxine in food. Nutritional yeast powder was used as a model food matrix. Acid extraction was compared with various enzymatic treatments in ammonium formate buffer to find a suitable method for the conversion of more complex vitamers into the same forms as the used isotope-labeled internal standards. The enzyme preparations alpha-amylase, takadiastase, beta-glucosidase, and acid phosphatase were all able to liberate thiamine and riboflavin. The diastatic enzyme preparations alpha-amylase and takadiastase also expressed proteolytic side activities resulting in the formation of small peptides which interfered with the mass spectra of thiamine and riboflavin. Liberation of nicotinamide and pantothenic acid from NAD(+) and CoA, respectively, could not be achieved with any of the studied enzyme preparations. Hydrochloric acid extraction at 121 A degrees C for 30 min was found to be destructive to pantothenic acid, but increased the liberation of pyridoxal.