HPLC同时测定苦黄注射液中生物碱和大黄蒽醌类成分的含量

Determination of Alkaloid and Rhubarb Anthraquinones Constituents in Kuhuang Injection by HPLC

目的建立同时测定苦黄注射液中生物碱和大黄蒽醌类成分的定量分析方法。方法采用Venusil MP C_(18)(2)(4.6 mm×250 mm,5μm)色谱柱为分析柱,甲醇(A)-0.4%二乙胺(B),梯度洗脱(A为5%→5%→40%→60%→80%→5%,相应时间周期为0→5→20→30→55→60 min),流速为1.0 m L·min^(-1),紫外检测波长为254 nm,柱温位30℃。结果在选定色谱条件下,芦荟大黄素、大黄酸、苦参碱、大黄素、大黄酚、氧化苦参碱、大黄素甲醚、槐定碱、槐果碱分别在0.012 3~0.369μg、0.023~0.69μg、0.138~4.14μg、0.056~1.68μg、0.065~1.95μg、0.145~4.35μg、0.012 5~0.375μg、0.146~4.38μg和0.101~3.03μg内线性关系良好(r=0.999 3,0.999 6,0.999 6,0.999 6,0.999 4,0.999 3,0.999 5,0.999 4,0.999 4),加样回收率分别为98.38%,100.79%,98.03%,99.81%,98.62%,99.28%,98.90%,101.75%,100.75%,RSD分别为1.60%,2.65%,2.12%,2.78%,2.08%,2.45%,2.08%,2.35%,1.72%。结论该方法操作简便易行,重复性好,结果准确可靠,可为苦黄注射液的质量控制提供定量评价方法。

OBJECTIVE To carry out the quantitative determination of alkaloid and Rhubarb anthraquinones constituents in Kuhuang injection by HPLC. METHODS HPLC-UV was used to determine alkaloid and Rhubarb anthraquinones constituents in Kuhuang injection. The separation was performed on Venusil MP C18（2） column（4.6 mm×250 mm, 5 μm） with gradient elution using methanol（A）-0.4% diethylamine（B） as mobile phase, gradient elute was that A for 5%, 5%, 40%, 40%, 80%, 60%, when the corresponding time period was 0, 5, 20, 30, 55, 60 min, and the detection wavelength was set at 254 nm. The flow rate was 1.0 mL·min^-1. The column temperature was 30 ℃. RESULTS The linear relationship of aloe emodin, rhein, matrine, emodin, rhubarb phenol, oxymatrine, emodin methyl ether, sophoridine and sophocarpine were within the range of 0.012 3-0.369 μg, 0.023-0.69 μg, 0.138-4.14 μg, 0.056-1.68 μg, 0.065-1.95 μg, 0.145-4.35 μg, 0.012 5-0.375 μg, 0.146-4.38 μg and 0.101-3.03 μg. The correlation coefficient were 0.999 3, 0.999 6, 0.999 6, 0.999 6, 0.999 4, 0.999 3, 0.999 5, 0.999 4, 0.999 4, respectively. The average recoveries were 98.38%, 100.79%, 98.03%, 99.81%, 98.62%, 99.28%, 98.90%, 101.75%, 100.75%, respectively. And RSD were 1.60%, 2.65%,2.12%, 2.78%, 2.08%, 2.45%, 2.08%, 2.35%, 1.72%, respectively. CONCLUSION The HPLC method is reproducible, feasible and precise. It can be used for the quality control of Kuhuang injection.