基于物质基础的雪上一枝蒿饮片及其水提物体内安全性比较研究  

Comparative Study on Safety of Processed Aconitum brachypoaum Diels.and Its Aqueous Extract in Vivo Based on Material Basis

作　　者：杨宏博 曹红云 张柏娥 李志玲 柳启涛 那敏 泰映玲 王京昆 YANG Hongbo;CAO Hongyun;ZHANG Baie;LI Zhiing;LIU Qitao;NA Min;TAI Yinging;WANG Jingkun(Yunnan Institute of Materia Medica,Yunnan Province Company Key Laboratory for TCM and Ethnic Drug of New Drug Creation,Kunming 650111,China)

机构地区：[1]云南省药物研究所,云南省中药和民族药新药创制企业重点实验室,云南昆明650111

出　　处：《中国中医药信息杂志》2024年第5期125-130,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：云南省重大科技专项(生物医药)课题(20181105)。

摘　　要：目的建立HPLC同时检测雪上一枝蒿饮片6个乌头类生物碱含量的方法,比较饮片及其水提干膏粉的急性毒性反应特征和严重程度差异,为雪上一枝蒿饮片质量标准提高及合理使用提供依据。方法采用ThermoAcclaim 120 C18色谱柱,以乙腈-四氢呋喃(5∶3,V/V)和0.1 mol/L醋酸铵溶液为流动相,梯度洗脱,柱温30℃,流速1.0 mL/min,测定雪上一枝蒿饮片及其水提干膏粉6个生物碱含量。将100只健康ICR小鼠随机分为溶媒对照组(0.5%羧甲基纤维素钠溶液),雪上一枝蒿饮片6.50、7.65g/kg组及水提干膏粉10.33、12.15 g/kg组,给予相应药物后观察小鼠14 d内的中毒表观特征及死亡情况。结果雪上一枝蒿炮制饮片及其水提干膏粉中苯甲酰乌头原碱的含量为0.10%和0.42%,其余5个生物碱均未检出。毒性试验结果表明,雪上一枝蒿饮片小鼠最大耐受剂量为6.50g/kg(相当于临床人拟用剂量的7831倍),其水提干膏粉小鼠最小致死剂量为30.99g/kg(相当于临床人拟用剂量的193688倍)。结论本研究建立的HPLC同时测定雪上一枝蒿饮片及其水提干膏粉6个乌头类生物碱含量的方法线性关系良好且准确性高,雪上一枝蒿饮片水提干膏粉乌头类生物碱含量更高且安全性更好。Objective To establish an HPLC method for simultaneous detection of six aconitum alkaloids in processed Aconitum brachypodum Diels.;To compare feature of the acute toxicity reaction and severity of the decoction pieces and its aqueous extract;To further provide a theoretical support for rising quality standards and reasonable use of Aconitum brachypodum Diels.Methods Six kinds of aconitum alkaloids of the decoction pieces and its aqueous extract of Aconitum brachypodum Diels.were separated on a Thermo Acclaim 120 C18 column,eluted gradually with mobile phase of acetonitrile-tetrahydrofuran(5∶3,V/V)and 0.1 mol/L ammonium acetate,with detected flow rate at 1.0 mL/min and column temperature at 30℃.100 ICR mice were randomly divided into 5 groups,i.e.control group(0.5%CMC-Na solution),two processed Aconitum brachypodum Diels.groups in doses of 6.50 g/kg and 7.65 g/kg,two aqueous extract groups of processed Aconitum brachypodum Diels.in doses of 10.33 g/kg and 12.15 g/kg.Apparent symptoms and deaths of mice were observed every day in 14 days after administration of corresponding drugs.Results Benzoyl aconitine contents which detected in processed Aconitum brachypodum Diels.and its aqueous extract,were 0.10%and 0.42%,respectively,as well as other five aconitines were not detected in test substances.The results of toxicity test showed that the maximum tolerate dose(MTD)of processed Aconitum brachypodum Diels.in mice was 6.50 g/kg(equivalent to 7831 times of proposed clinical dosage in human),while the minimum lethal dose(MLD)of its aqueous extract was 30.99 g/kg(equivalent to 193688 times of proposed clinical dosage in human).Conclusion The established HPLC method for simultaneous detection of six kinds of aconitum alkaloids in decoction pieces and its aqueous extract of Aconitum brachypodum Diels.is well linear and accurate.Compared to processed Aconitum brachypodum Diels.,its aqueous extract with higher aconitum alkaloids content has a better safety.

关 键 词：雪上一枝蒿 炮制 物质基础 乌头类生物碱 急性毒性 安全性 

分 类 号：R284.1[医药卫生—中药学] R285.5[医药卫生—中医学]