反相高效液相色谱法同时测定地黄中5种成分的含量

Simultaneous Determination of Five Components in Rehmanniae Radix by HPLC

目的建立地黄药材中梓醇、益母草苷、腺苷、肉苁蓉苷A及毛蕊花糖苷等5种成分的含量测定方法。方法采用HPLC,色谱柱为Grace Allitma C18(4.6 mm×250 mm,5μm),以乙腈-水为流动相,线性梯度洗脱,流速为1.0 mL.min-1,检测波长为203 nm,柱温为25℃。结果梓醇、益母草苷、腺苷、肉苁蓉苷A和毛蕊花糖苷质量浓度分别在12.47～997.22、2.85～227.78、0.36～28.75、1.30～104.17、1.65～131.94μg.mL-1内呈良好线性关系,相关系数分别为0.999 7、1.000 0、0.999 9、0.999 9和0.999 7,平均回收率分别为99.7%(RSD=2.20%,n=9)、98.8%(RSD=2.38%,n=9)、98.8%(RSD=3.10%,n=9)、99.1%(RSD=3.28%,n=9)和100.0%(RSD=2.10%,n=9)。结论该方法简便、准确、重现性好,可用于地黄的质量控制。不同产地地黄药材中5种指标成分含量差异较大。

OBJECTIVE To develop an HPLC method for simultaneous determination of catalpol, leonuride, adenosine, cistanoside A and acteoside in Rehmanniae Radix. METHODS The HPLC analysis was carried out on a Grace Allitma Cls column （4. 6 mm ×250 mm,5 μm）, with a linear gradient elution of acetonitrile-water at the flow rate of 1.0 mL·min-1. The detective wave length was set at 203 nm, and the column temperature was set at 25 ℃. RESULTS The calibration curves were linear within the range of 12.47 -997.22 μg·mL-1 for catalpo], 2. 85 -227.78μg·mL-1 for leonuride, 0. 36 -28.75 μg·mL-1 for adenosine, 1.30 - 104. 17 μg·mL-1 for cistanoside A, and 1.65 - 131.94μg·mL-1 for aeteoside, respectively. The average recoveries for the five marker compounds were 98.8% - 100. 0% , r = 0. 999 7 - 1. 000 0. CONCLUSION This method is simple, accurate and practical for the quality control of Rehmanniae Radix. There are some differences in the contents of the five marker compounds in the samples from different producing areas.