HPLC-DAD法检测桑叶中的1-脱氧野尻霉素

Determination of 1-deoxynojirimycin in mulberry leaves by HPLC-DAD

本实验旨在建立检测桑叶中1-脱氧野尻霉素(1-DNJ)的高效液相色谱(HPLC)-二极管阵列检测器(DAD)方法。以0.05mol/LHCl为提取溶剂,芴甲氧羰酰氯(FMOC-Cl)为衍生剂,在pH8.5的硼酸盐缓冲液中进行衍生化反应,生成具有紫外吸收的1-DNJ-FMOC络合物,采用C18色谱柱,乙腈和水为流动相,最大吸收波长为265nm处对目标物进行检测。结果表明:1-DNJ的色谱图分离效果良好,线性相关系数为0.9997,检出限为0.015mg/mL。样品平均加标回收率为81.3%~92.1%,相对标准偏差(RSDs)为4.7%~6.5%(n=5)。

1-deoxynojirimycin (1-DNJ) is a special multi-hydroxyl alkaloid contained in the genus Morus,which is the main active ingredient of mulberry leaf lowering blood sugar.A high performance liquid chromatography(HPLC)-diode array detector (DAD)method was developed for determination of 1-DNJ in mulberry leaves.Taking 0.05 mol/L HCl as the extraction solvent,FMOC-Cl was the derivative agent,the extract was derived in the borate buffer (pH=8.5)to generate the 1-DNJ-FMOC with UV absorption.Under maximum absorption wavelengths of the studied analytes was 265 nm,the 1-DNJ detected by C18 column with acetonitrile and water as mobile phase.The results showed that 1-DNJ chromatography was suitable for separation,the linear correlation coefficients was 0.9997,the detection limits was 0.015 mg/mL,the average spiked recoveries was 81.3%~ 92.1%,and the relative standard deviations(RSDs)was 4.7%~ 6.5%(n=5).