高效液相色谱法同时测定复方褪黑激素片中维生素B_6与褪黑激素的含量

Simultaneous determination of vitamin B_6 and melatonin in Compound Melatonin Tablets by high performance liquid chromatography

目的应用高效液相色谱(HPLC)技术,建立同时测定复方褪黑激素片中维生素B_6和褪黑激素含量的方法。方法采用Alltima C_(18)色谱柱(150 mm×4.6 mm,5μm),以50 mmol/L磷酸二氢铵溶液(磷酸调至pH 3.0)-乙腈为流动相,梯度洗脱,流速0.5 m L/min,检测波长278 nm,柱温30℃,进样体积10μL,对复方褪黑激素片中的维生素B_6和褪黑激素进行含量测定。结果维生素B_6和褪黑激素检测浓度分别在80.34~401.7 mg/L(r=0.9997)和8.50~42.5 mg/L(r=0.9999)范围内与峰面积呈良好的线性关系;精密度和稳定性良好,相对标准偏差(RSD)均<0.8%;回收率良好,维生素B6和褪黑激素的平均回收率(n=9)分别为100.4%和98.86%,RSD分别为0.42%和0.86%。复方褪黑激素片中维生素B_6平均含量为14.14 mg/片,褪黑激素平均含量为0.9853 mg/片。结论该方法准确、简单、快速,可用于同时测定复方褪黑激素片中维生素B_6和褪黑激素的含量。

Objective To develop a high performance liquid chromatography （HPLC） method for the simultaneous de termination of vitamin B6 and melatonin in Compound Melatonin Tablets. Methods An Atltima C18 column （150 mm × 4.6 mm, 5 μm） was used for the separation, with gradient elution of 50 mmol/L ammonium dihydrogen phosphate （adjusted pH to 3.0 with HaPO4） and acetonitrile as the mobile phase at the flow rate of 0.5 mL/min. The detection wavelength was set at 278 nm. The column temperature was 30~C and injection volume was 10 μL. Vitamin B6 and melatonin in Compound Melatonin Tablets were quantified by the established method. Results The liner range of vitamin B6 and melatonin was 80.34-401.7 mg/L （r=0.9997） and 8.50-42.5 mg/L （r=0.9999） respectively. The precision and stability were satisfactory, and the relative standard deviation （RSD） were less than 0.8%. The average recoveries （n=9） was 100.4% （RSD=0.42%） and 98.86% （RSD=0.86%） for vitamin B6 and melatonin respectively. The average contents of vitamin B6 and melatonin in Compound Melatonin Tablets were 14.14 mg/tablet and 0.9853 mg/tablet. Conclusion The established method is accurate, simple and rapid, and can be utilized for simultaneous determination of vitamin B6 and melatonin in Compound Melatonin Tablets.