Li Ke
The Center of Medical Laboratory Science, Jinling Hospital, Clinical School of Medical College, Nanjing University, No. 305 East Zhongshan Road, Jiangsu, People's Republic of China.
Biomed Chromatogr. 2002 Dec;16(8):504-7. doi: 10.1002/bmc.192.
A reversed-phase ion-pair high performance liquid chromatographic method (HPLC) has been developed and validated for the routine analysis of nicotinamide, pyridoxine hydrochloride, thiamine mononitrate and riboflavin in multivitamin with minerals tablets. HPLC separation of the vitamins was performed on a Hypersil C(18) column and detected by ultraviolet absorbance at 280 nm. The use of methanol-aqueous 0.5% acetic acid solution (18:82, v/v; containing 2.5 mM sodium hexanesulfonate, pH = 2.8) as the mobile phase at a flow-rate of 1.2 mL/min enables the baseline separation of the four analytes free from interferences with isocratic elution at 30 degrees C. The analysis time was 17 min per injection. The method was linear in the ranges of 5-90, 2.5-90, 5-95 and 25-450 micro g/mL for thiamine mononitrate, riboflavin, pyridoxine hydrochloride and nicotinamide, respectively. The average coefficients of variation of within- and between-day assays were 2.2 and 3.6% for thiamine mononitrate, 1.8 and 2.4% for riboflavin, 1.3 and 1.7% for pyridoxine hydrochloride and 1.0 and 1.5% for nicotinamide, respectively. The average recoveries of thiamine mononitrate, riboflavin, pyridoxine hydrochloride and nicotinamide were 97.0, 97.2, 98.9 and 100.4% for the tablets, respectively. The method has been successfully applied to the simultaneous determination of nicotinamide, pyridoxine hydrochloride, thiamine mononitrate and riboflavin in multivitamin with minerals tablets.
已开发并验证了一种反相离子对高效液相色谱法(HPLC),用于常规分析多种维生素矿物质片中的烟酰胺、盐酸吡哆醇、硝酸硫胺素和核黄素。维生素的HPLC分离在Hypersil C(18)柱上进行,并通过280 nm处的紫外吸光度进行检测。以甲醇 - 0.5%乙酸水溶液(18:82,v/v;含有2.5 mM己烷磺酸钠,pH = 2.8)作为流动相,流速为1.2 mL/min,可在30℃下等度洗脱,实现四种分析物的基线分离,无干扰。每次进样的分析时间为17分钟。该方法对于硝酸硫胺素、核黄素、盐酸吡哆醇和烟酰胺的线性范围分别为5 - 90、2.5 - 90、5 - 95和25 - 450μg/mL。硝酸硫胺素日内和日间测定的平均变异系数分别为2.2%和3.6%,核黄素为1.8%和2.4%,盐酸吡哆醇为1.3%和1.7%,烟酰胺为1.0%和1.5%。硝酸硫胺素、核黄素、盐酸吡哆醇和烟酰胺在片剂中的平均回收率分别为97.0%、97.2%、98.9%和100.4%。该方法已成功应用于同时测定多种维生素矿物质片中的烟酰胺、盐酸吡哆醇、硝酸硫胺素和核黄素。