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目的:建立HPLC-UV法测定多维生素糖浆中D-生物素的含量。方法:样品用混合溶剂(丙酮:己烷:乙酸乙酯=1:2:1)萃取后,采用HPLC-UV法测定其中D-生物素的含量,使用Agela AQ-C18柱(250 mm×4.6 mm,5μm),流动相为5 mmol.L-1己烷磺酸钠溶液(以0.02 mol.L-1磷酸调pH至2.3)-乙腈(92∶8);流速为1.0 mL.min-1;检测波长为200 nm。结果:D-生物素的浓度在1~10μg.mL-1范围内与峰面积线性关系良好(r=0.999 9,n=7);平均回收率为100.7%(RSD=1.4%,n=6)。结论:本实验所建立的HPLC-UV分析方法快速、灵敏,色谱峰分离度良好,结果准确可靠,可作为多维生素糖浆的质量控制项目之一。
Objective: To establish HPLC-UV method for the determination of D-biotin in multivitamin syrup. Methods: The sample was extracted with mixed solvent (acetone: hexane: ethyl acetate = 1: 2: 1), and the content of D-biotin was determined by HPLC-UV method using Agela AQ-C18 column The mobile phase consisted of 5 mmol·L-1 hexanesulfonate solution (adjusted to pH 2.3 with 0.02 mol·L-1 phosphate) -acetonitrile (92: 8) at a flow rate of 1.0 mL · min-1 ; Detection wavelength of 200 nm. Results: The concentration of D-biotin was linear with the peak area in the range of 1 ~ 10μg.mL-1 (r = 0.999 9, n = 7). The average recovery was 100.7% (RSD = 1.4%, n = 6 ). Conclusion: The HPLC-UV analysis method established in this experiment is rapid, sensitive and has good chromatographic peak resolution. The results are accurate and reliable and can be used as one of the quality control items for multi-vitamin syrup.