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

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．