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目的:采用HPLC法建立大黄中4种蒽醌类有效成分的含量测定方法。方法:采用色谱柱:Agi-lent C18柱(250mm×4.6mm,5μm);流动相:A为甲醇,B为0.5%磷酸水溶液,C为乙腈,洗脱梯度:0～5min(9%A,80%B,11%C),5～20min(9%～8%A,80%～72%B,11%～20%C),20～35min(8%～7%A,72%～60%B,20%～33%C),35～50min(7%～5%A,60%～50%B,33%～45%C),50～60min(5%～4%A,50%～36%B,45%～60%C),60～80min(4%～0%A,36%～9%B,60%～91%C);流速:1mL.min-1;检测波长:280nm;柱温:25℃。结果:4种蒽醌在80min内即可达到完全分离,芦荟大黄素、大黄酸、大黄素和大黄素甲醚分别在0.97～19.40μg.mL-1、1.08～21.60μg.mL-1、0.97～19.40μg.mL-1、1.60～32.00μg.mL-1范围内线性关系良好。大黄中大黄酸、大黄素在60%乙醇作为溶剂提取率最高。结论:提取溶剂不同对大黄饮片中蒽醌成分含量变化有影响。
OBJECTIVE: To establish a HPLC method for the determination of four anthraquinones in rhubarb. The mobile phase consisted of methanol A, B 0.5% phosphoric acid aqueous solution, acetonitrile C as elution gradient 0-5 min (9% A, 80% B, 11% C), 5-20 min (9% -8% A, 80% -72% B, 11% -20% C), 20-35 min % B, 20% -33% C), 35-50 min (5-7% A, 60-50% B, 33-45% C) ~ 36% B, 45% ~ 60% C), 60 ~ 80min (4% ~ 0% A, 36% ~ 9% B, 60% ~ 91% 280 nm; column temperature: 25 ° C. Results: Four kinds of anthraquinones could be completely separated within 80 minutes. The contents of aloe-emodin, rhein, emodin and physcion were 0.97 ~ 19.40μg.mL-1, 1.08 ~ 21.60μg.mL-1, 0.97 ~ 19.40μg.mL-1, 1.60 ~ 32.00μg.mL-1 in the range of good linear relationship. Rhein, rhein, emodin in 60% ethanol as the highest solvent extraction rate. Conclusion: Different extraction solvents affect the content of anthraquinone in rhubarb decoction.