止痛温经散中乌头碱经皮给药的血药浓度测定及安全性分析

Blood Concentration Determination of Aconitine in ERON Powder by Transdermal Delivery and Security Analysis

目的:探究止痛温经散中乌头碱经皮给药的血药浓度的测定及安全性。方法:将12只小鼠平均分为对照组与敷药组,对照组不进行处理,敷药组小鼠背部敷用止痛温经散膏状制剂,采用液相色谱-质谱联用仪(LC-MS)对给药后2h、4h、6h、12h、24h、48h血浆中乌头碱浓度进行测定;另选取18只小鼠分为三组,每组6只,在小鼠脊背左侧皮肤敷含有不同浓度的止痛温经散(7.5mg/g、15mg/g、30mg/g),每只小鼠均以右侧皮肤作为对照,对1h、24h、48h、72h皮肤红斑、水肿情况进行评分;采用标准II导联仪记录小鼠给药1h、2h、4h、6h心电图,以给药前测定为基线值,记录小鼠给药后心率、QT间期、QRS间期变化。结果:乌头碱线性方程为y=0.243x-0.007(r=0.998),在1ng^500ng范围内线性关系良好,1、20、40ng/ml乌头碱标准品回收率分别为(94.53±4.07)%、(96.43±5.73)%、(93.20±3.68)%,稳定性RSD均<5%,重复性RSD均<1。经皮给药后,在2~24h内随着时间延长,小鼠血浆中乌头碱浓度逐渐增高,48h浓度降低,在24h时浓度达到最大值(P<0.05)。低剂量草乌细粉处理时,不同时间点皮肤得分对比,差异无统计学意义(P>0.05)。中剂量浓度处理时,1h给药侧皮肤得分高于对照侧,高剂量止痛温经散处理后,不同时间点给药侧皮肤得分均高于对照侧,且高于低、中剂量皮肤得分,差异有统计学意义(P<0.05)。经皮给药后,小鼠心率无明显变化,QRS间期整体呈现不规律缩短,在给药6h后QT间期变化最大。结论:止痛温经散经皮给药后,乌头碱成分能够通过皮肤较好的吸收,连续敷用能够较好维持血浆中药物浓度,对皮肤刺激性小、心率变化较小。

Objective： To explore blood concentration determination of aconitine in ERON powder by transdermal delivery and analyze the security. Methods： 12mice were divided into controlled group and treated group, the controlled group received no treatment and were treated mice back apply ointment preparation temperature analgesic powder, ERON ointment was applied to the back of a mouse, and liquid chromatography mass spectrometry （LC-MS） was used to detect plasma aconitine concentration after administration of 2h, 4h, 6h, 12h, 24h and 48h. ERON ointment, with different concentrations of ERON powder （7.5mg/g, 15mg/g, 30mg/g）, each mouse was treated with the right skin as the control. Another 18mice were divided into three groups, 6rats in each group, the right skin was used as a control, and skin erythema and edema of lh, 24h, 48h, 72h were scored. Standard II lead instrument was used to recorded electrocardiogram at administration of lh, 2h, 4h, 6h. The baseline value was determined before administration, and the changes of heart rate, QT interval and QRS interval were recorded after administration. Results ： Aconitine linear equation was that y = 0. 243x-0. 007 （ r = 0, 998）, with good linear relations in lng -500ng. Aconitine standard product recovery rates of 1, 20, 40ng/ml were （94. 53±4. 07）%, （96. 43±5.73）%, and （93.20±3.68） %, the stability of RSD was less than 5%, and the repeatability of RSD was less than 1. After transdermal ad- ministration, as time extended in 2- 24h, the concentration of aconitine increased gradually in mice plasma, and at 48h de- creased, and the concentration reached the maximum at 24h （P〈0. 05）. In treatment of low-dose aconitum powder, there were no statistically significant differences in skin score at different time points （ P〉0. 05 ）. In treatment of median-dose aconitum powder, skin score in the administration side was higher than that in the controlled side. In treatment of high-dose aconitum powder, skin scores in the administration side at different time points were higher than those in the controlled side, higher than those of low-dose and median-dose aconitum powde, with statistically significant difference （P〈0. 05）. After percutaneous ad- ministration, it showed no changes of heart rate of the mice, and QRS interval whole showed irregular shortening, and QT interval changed reached to the maximum after administration of 6h. Conclusion： After transdermal administration of ERON powder, aconitine can be better absorbed by the skin, and continuous application can better maintain plasma concentration of drugs, with small skin irritation and heart rate change.