HPLC法测定化橘红中水合橘皮内酯、橘皮内酯、马尔敏和葡萄内酯的含量

目的: 建立HPLC法同时测定化橘红药材中水合橘皮内酯、橘皮内酯、马尔敏和葡萄内酯的含量。方法: 采用DiamonsilC₁₈ 色谱柱(250 mm×4.6 mm,5 μm),以甲醇(A)-水(B)为流动相,二元梯度洗脱(0~6 min,49%A;6~15 min,49%A→75%A;15~26 min,75%A→100%A;26~30 min,100%A;30~32 min,100%A→49%A;32~35 min,49%A),流速1.0 mL · min^-1,检测波长324 nm,柱温30 ℃。结果: 水合橘皮内酯、橘皮内酯、马尔敏和葡萄内酯的进样量分别在0.378 0~2.268 μg(r=0.999 7)、0.088 56~0.531 4 μg(r=0.999 7)、0.089 58~0.537 5 μg(r=0.999 8)和0.053 38~0.320 3 μg(r=0.999 9)范围内呈良好的线性关系;平均回收率(n=6)分别为100.6%、101.5%、99.68%、98.93%,RSD为2.9%、2.7%、2.2%、2.3%。12批化橘红样品中水合橘皮内酯、橘皮内酯、马尔敏和葡萄内酯含量测定结果分别为0.123%~0.452%、0.036%~0.094%、0~0.090%、0~0.035%。结论: 经方法学验证,本法可用于化橘红药材中水合橘皮内酯、橘皮内酯、马尔敏和葡萄内酯的含量测定。

Objective: To develop an HPLC method for simultaneous determination of meranzin hydrate, meranzin, marmin and aurapten in Citri Grandis Exocarpium. Methods: The HPLC system consisted of a Diamonsil C₁₈ column(250 mm×4.6 mm, 5 μm)and the mobile phase consisted of methanol(A)-water(B)in gradient mode(0-6 min, 49%A;6-15 min, 49%A→75%A;15-26 min, 75%A→100%A;26-30 min, 100%A;30-32 min, 100%A→49%A;32-35 min, 49%A)at the flow rate of 1.0 mL · min^-1.The detection wavelength was 324 nm and the column temperature was 30 ℃. Results: The contents of meranzin hydrate, meranzin, marmin and aurapten had good linear relationship in the ranges of 0.378 0-2.268 μg(r=0.999 7), 0.088 56-0.531 4 μg(r=0.999 7), 0.089 58 -0.537 5 μg(r=0.999 8)and 0.053 38-0.320 3 μg(r=0.999 9)respectively, and the average recoveries(n=6)of four components were 100.6% with RSD of 2.9%, 101.5% with RSD of 2.7%, 99.68% with RSD of 2.2% and 98.93% with RSD of 2.3% respectively.The contents in 12 samples of Citri Grandis Exocarpium were 0.123%-0.452% for meranzin hydrate, 0.036%-0.094% for meranzin, 0-0.090%for marmin, and 0-0.035% for aurapten. Conclusion: According to the methodology validation, this method can be applied to the determination of meranzin hydrate, meranzin, marmin and aurapten in Citri Grandis Exocarpium.

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