目的:建立HPLC 法同时测定厚朴三物汤中芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚、和厚朴酚、厚朴酚的含量。方法:采用Diamonsil C18 色谱柱(250 mm×4.6 mm,5 μm),流动相为甲醇(A)-0.1％ 磷酸水(B),二元梯度洗脱(0~15 min,64%A;15~23 min,64%A → 75%A;23~33 min,75%A;33~47 min, 75%A → 90%A;47~55 min,90%A),检测波长为 294 nm,流速1.0 mL·min-1,柱温25 ℃ ,进样量20 μL; 同时进行方法学验证。结果:芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚、和厚朴酚、厚朴酚的进样量范围分别为40.18~1 607.2 ng(r=0.999 9)、38.06~1 522.4 ng(r=0.999 6)、35.9~1 436 ng(r=0.999 8)、65.86~2634.4 ng(r=0.999 8)、28.74~1149.6 ng(r=0.999 7)、24~964 ng(r=0.999 9)和48.3~1932 ng(r=0.999 8),与峰面积呈良好的线性关系;平均回收率(n=6)分别为96.4%、98.9%、99.0%、101.0%、100.8%、97.3%、98.8%,RSD 为1.5%、1.8%、2.0%、2.0%、1.4%、2.4%、1.1%;实测3 个厚朴三物汤中7 个成分含量结果分别为： 芦荟大黄素0.102 1~0.128 4 mg·mL-1、大黄酸0.147 2~0.184 0 mg·mL-1、大黄酚0.107 7~0.126 8 mg·mL-1、大黄素0.191 9~0.256 6 mg·mL-1、大黄素甲醚0.091 1~0.104 6 mg·mL-1、和厚朴酚0.031 7~0.035 2 mg·mL-1、厚朴酚0.093 4~0.106 2 mg·mL-1。结论:建立的方法准确、简便、灵敏,可用于厚朴三物汤中芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚、和厚朴酚、厚朴酚的含量测定及质量控制研究。
Objective: To develop an HPLC method for simultaneous determination of aloe-emodin,rhein, emodin,chrysophanol,physcion,honokiol and magnolol in Houposanwu decoction. Methods: A Diamonsil C18 column(250 mm×4.6 mm,5 μm) was adopted with the mobile phase of methanol(A)-0.1% phosphoric acid water(B) in gradient mode(0-15 min,64%A;15-23 min,64%A → 75%A;23-33 min,75%A;33-47 min,75%A → 90%A;47-55 min,90%A). The detection wavelength was 294 nm,the flow rate was 1.0 mL min-1,the column temperature was 25 ℃ and injection volume was 20 μL at the same time,the reliability of the method was validated. Results: The contents of aloe-emodin,rhein,emodin,chrysophanol,physcion,honokiol and magnolol had good linear relationship in the ranges of 40.18-1 607.2 ng(r=0.999 9),38.06-1 522.4 ng(r=0.999 6),35.9-1 436 ng(r=0.999 8),65.86~2 634.4 ng(r=0.999 8),28.74-1149.6 ng(r=0.999 7),24-964 ng(r=0.999 9) and 48.3-1 932 ng(r=0.999 8),respectively,and the average recoveries(n=6) of seven components were 96.4% with RSD of 1.5%,98.9% with RSD of 1.8%,99.0% with RSD of 2.0%,101.0% with RSD of 2.0%,100.8% with RSD of 1.4%,97.3% with RSD of 2.4% and 98.8% with RSD of 1.1%,respectively. The contents of 7 components in 3 Houposanwu were measured and the results were as following:aloe-emodin 0.102 1-0.128 4 mg·mL-1,rhein 0.147 2-0.184 0 mg·mL-1,chrysophanol 0.107 7-0.126 8 mg·mL-1,emodin 0.191 9-0.256 6 mg·mL-1,physcion 0.091 1-0.104 6 mg·mL-1,honokiol 0.031 7-0.035 2 mg·mL-1 and magnolol 0.093 4-0.106 2 mg·mL-1. Conclusion: The method is simple,accurate and reliable,and can be used as an effective method for the determination of aloe-emodin,rhein,emodin,chrysophanol,physcion,honokiol and magnolol in Houposanwu(decoction),and it can also provide the basis for the quality control research.
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