食品科学

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钛胶RP-HPLC法同时测定5 种维生素

耿 瑛,李 荣,姜子涛,张发博   

  1. 天津商业大学生物技术与食品科学学院,天津市食品生物技术重点实验室,天津 300134
  • 出版日期:2016-07-25 发布日期:2016-07-26
  • 通讯作者: 李 荣
  • 基金资助:

    天津市自然科学基金项目(13JCYBJC18700;12JCZDJC34100)

Simultaneous Determination of Five Vitamins by Titania-Based RP-HPLC

GENG Ying, LI Rong, JIANG Zitao, ZHANG Fabo   

  1. Tianjin Key Laboratory of Food Biotechnology, College of Biotechnology and Food Science,
    Tianjin University of Commerce, Tianjin 300134, China
  • Online:2016-07-25 Published:2016-07-26
  • Contact: LI Rong

摘要:

采用钛胶反相色谱柱,建立高效液相色谱法同时测定5 种维生素:烟酸、生物素、烟酰胺、叶酸和钴胺素含量的检测方法。研究5 种维生素的保留行为与磷酸缓冲盐pH值、缓冲盐类型及浓度、柱温和流速的关系,得出了最优色谱条件:于波长210 nm处检测生物素和钴胺素,于波长270 nm处检测烟酸、烟酰胺和叶酸,磷酸盐pH 7.0,浓度5.0 mmol/L,柱温50 ℃,流速0.8 mL/min。分析5 种维生素保留的热力学参数焓、熵和吉布斯自由能。烟酸、生物素、烟酰胺、叶酸和钴胺素标准曲线线性良好(R2>0.999 0);烟酸、生物素、烟酰胺、叶酸和钴胺素的检出限分别为6、10、20、22、8 ng/mL;5 种物质的相对标准偏差均低于1.36%;加标回收率范围为92.30%~107.20%。方法的精密度和准确度均可满足高效液相色谱法的测定要求。

关键词: 钛胶反相色谱柱, 高效液相色谱法, 维生素

Abstract:

A new method for simultaneous separation and determination of five vitamins including nicotinic acid (vitamin
PP), biotin (VB7), nicotinamide (vitamin PP), folic acid (VB9), and cyanocobalamin (VB12) using high performance liquid
chromatography (HPLC) on a titania-based column has been developed. The influence of buffer pH, buffer type, buffer
concentration, column temperature and flow rate on separation efficiency was investigated. The optimized chromatographic
conditions were obtained as follows: 5.0 mmol/L phosphate solution at pH 7.0 as buffer solution, column temperature of
50 ℃, and a flow rate of 0.8 mL/min. Biotin and cyanocobalamin were detected at 210 nm and three other vitamins at
270 nm. The thermodynamic parameters enthalpy, entropy and Gibbs free energy were calculated for the retention of the
analytes. The proposed method presented good linearity (R2 > 0.999 0) for the five vitamins. The limits of detection (LODs)
for nicotinic acid, biotin, nicotinamide, folic acid and cyanocobalamin were 6, 10, 20, 22, and 8 ng/mL, respectively. The
precisions (RSDs) for the five vitamins were less than 1.36%. The recoveries of spiked samples were between 92.30% and
107.20%. The precision and accuracy of this method can meet the requirements of HPLC analysis.

Key words: titania-based reverse phase, high performance liquid chromatography (HPLC), vitamins

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