高效液相色谱法测定复方肝泰乐胶囊中维生素B_1、维生素B_2、烟酰胺和苯巴比妥含量

目的建立HPLC同时测定复方肝泰乐胶囊中维生素B1、维生素B2、烟酰胺和苯巴比妥含量的方法。方法色谱柱用十八烷基硅烷键合硅胶为固定相（150mm×4.6mm,4.6μm）。流动相A为乙腈-水-三乙胺（100：900：1,含0.01mol/L庚烷磺酸钠,用冰醋酸调pH值为3.6）,流动相B为乙腈。洗脱程序为0～12min,A100%～70%;12～200min,A70%;流速为1mL/min,检测波长为272nm。结果维生素B1、维生素B2、烟酰胺和苯巴比妥的保留时间分别约为4.7、10.1、12.4和14.2min,与各自相邻峰的分离度均大于1.5。以峰面积对进样量（μg）线性回归,维生素B1回归方程为y=186.83x＋17.85,线性范围为0.05332～0.34440μg,r=0.9999;维生素B2回归方程为y=402.47x＋12.54,线性范围为0.02511～0.19980μg,r=0.9999;烟酰胺回归方程为y=58.3141x＋206.8,线性范围为0.1641～1.265μg,r=0.9991;苯巴比妥回归方程为y=165.4x＋181.2,线性范围为0.5221～2.0030μg,r=0.9995。维生素B1、维生素B2、烟酰胺和苯巴比妥回收率分别为100.2%、100.1%、100.2%和100.2%,RSD分别为0.09%、0.09%、0.09%和0.08%。结论本方法精密度、准确度好,操作简便,可用于复方肝泰乐胶囊中维生素B1、维生素B2、烟酰胺和苯巴比妥含量测定。

Determination of vitamin B1, vitamin B2, nicotinamide and phenobarbital in the conpoumd Glucurolactone capsules by HPLC

Objective To develop an HPLC quantitative method for the determination of vitamin B1, vitamin B2, nicotinamide and phenobarbital in the compound Glucurolactone capsules. Methods The chromatographic conditions included C18 (150 mm×4.6 mm, 4.6 μm); mobile phase A was acetonitrile-water-triethylamine (100:900:1, contained 0.01 mol/L Sodium Heptane adjusted to pH 3.6 by acetic acid); mobile phase B was acetonitrile. Gradient elutiuon was 0-12 min,A100%-70%;12-200 min,A70%. The flow rate was 1 mL/min minand monitored at 272 nm. Results The retention time of vitamin B1, vitamin B2, nicotinamide and phenobarbital were 4.7 min, 10.1 min, 12.4 min and 14.2 min respectively. The separative degrees with each adjacent peak were bigger than 1.5. The regress equation for vitamin B1 was y = 186.83 x + 17.85, r = 0.999 9 and linear range was 0.053 32-0.344 40 μg; vitamin B2 y = 402.47 x + 12.54, r = 0.999 9 and linear range was 0.025 11-0.199 80 μg; nicotinamide y = 58.314 1 x + 206.8, r = 0.999 1 and linear range was 0.164 1-1.265 0 μg, Phenobarbital y = 165.4 x + 181.2, r = 0.999 5 and linear range was 0.522 1-2.003 0 μg. The average recovery of vitamin B1, vitamin B2, nicotinamide and phenobarbital were 0.09%, 0.09%, 0.09% and 0.08% respectively. Conclusion The method is sensitive, time-saving and accurate. The method can be used for measuring the contents of vitamin B1, vitaminB2, nicotinamide and phenobarbital in the compound Glucurolactone capsules.