Aim To develop an HPLC method for the determination of Aconitum alkaloids extracted from Radix aconiti preparata in rats. Methods Waters 2690@996 PAD system was used. The analytical column was a Halsil 100 C18 column (250 mm×4.6 mm ID, 5 μm). The mobile phase was water, methanol and diethyl amine at the ratio of 75∶ 25∶ 0.1. The flow rate was 0.9 mL·min -1. The wavelength of the detector was 240 nm. Results The linear ranges of aconitine in the heart, spleen, lung and kidney were 0.4-100 μg·mL -1, the correlation coefficients were 0.997 2, 0.998 6, 0.999 3 and 0.999 4, respectively. The linear range of aconitine in liver was 2-200 μg·mL -1 and the correlation coefficient was 0.999 0. The linear ranges of hypaconitine in heart, liver, spleen, lung, kidney, brain and spinal cord were 5-100 μg·mL -1, the correlation coefficients were 0.999 4, 0.999 7, 0.999 8, 0.998 4, 0.999 8, 0.999 8 and 0.999 7, respectively. Detection limits (S/N=3) of aconitine and hypaconitine were 0.4 μg·mL -1. The recoveries of aconitine and hypaconitine ranged from 88.7% to 102.2% and 86.5% to 101.3%, respectively, and the RSD of precision of aconitine and hypaconitine was 10%. Conclusion It appears to be an accurate and effective method that can offer reference basis for in toxication of Radix aconiti preparata clinically.%目的建立HPLC法测定口服制川乌提取物后大鼠体内乌头类生物碱的方法.方法用Waters 2690@996 PAD系统,Halsil 100 C18柱(250 mm×4.6 mm ID, 5 μm),水-甲醇-二乙胺(75∶ 25∶ 0.1)为流动相,流速0.9 mL·min -1,检测波长240 nm.结果乌头碱在心脏、脾脏、肺脏、肾脏的线性范围:0.4-100 μg·mL -1,r分别为0.997 2,0.998 6,0.999 3,0.999 4;在肝脏的线性范围:2-200 μg·mL -1,r为0.999 0.次乌头碱在心、肝、脾、肺、肾、脑和脊髓的线性范围:5-100 μg·mL -1,r分别为0.999 4,0.999 7,0.999 8,0.998 4,0.999 8,0.999 8和0.999 7.乌头碱及次乌头碱的检测限(S/N=3)为0.4 μg·mL -1.各组织中的回收率:乌头碱为88.7%-102.2%,次乌头碱为86.5%-101.3%.结论本法为确证乌头类生物碱的中毒提供了科学有效的依据.
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