应用UPLC-Orbitrap-MS分析草乌及其不同炮制品化学成分差异

草乌是有毒药之一,需经过炮制后方能使用。中药中草乌常用的炮制方法为水煮至内无白心,口尝微有麻舌感,蒙古族药中常用诃子制草乌,2种方法均能减毒。该实验利用UPLC-Orbitrap-MS对生草乌、药典法制草乌、诃子制草乌进行分析,比较炮制前后化学成分变化,为阐明诃子制草乌炮制减毒原理提供依据。通过比对生草乌与各炮制品的正负离子模式离子流出峰分析草乌炮制前后成分变化情况,并以化合物炮制前后离子峰面积比为炮制前后变化指数,将草乌主要活性成分经2种炮制方法炮制后的含量变化趋势进行对比,发现生草乌经药典法炮制后,新增14-O-anisoylneoline,dehydro-mesaconitine等14种成分,同时N-demethyl-mesaconitine,aconitine消失,且双酯型生物碱含量明显减少,单酯型和醇胺型生物碱含量明显增加,说明药典法制草乌是通过降低主要毒性成分双酯型生物碱的含量来减毒的;草乌经诃子汤炮制后,原草乌中含有的生物碱类化学成分变化较小,新增gallic acid,chebulic acid,shikimic acid等9种诃子中含有的化学成分,猜测诃子制草乌的减毒机制可能与诃子中化学成分有关。该研究结论表明药典法制草乌与诃子制草乌中化学成分有质的不同,二者具有不同的减毒原理。

Chemical variation in Aconti Kusnezoffii Radix before and after processing based on UPLC-Orbitrap-MS

Some Chinese herbal medicine needs to be processed before it can be used as medicine,especially toxic Chinese medicine.Highly toxic Aconti Kusnezoffii Radix(Caowu in Chinese)is widely used in traditional Chinese medicine and Mongolian medicine.In traditional Chinese medicine,Caowu is usually processed by boiling with water(CW)until no white part inside and being tasted without tongue-numbing.In Mongolian medicine,it is usually soaked in Chebulae Fructus(Hezi in Chinese)decoction for several days(CH).Both methods could reduce toxicity according to reports.The biggest difference between CW and CH is that CW needs to be heated for 4-6 h,while CH needs Hezi as processing adjuvants.To explore the toxicity reduction mechanism of CW and CH,we studied the contents of various compounds in Caowu processed by two methods by UPLC-Orbitrap-MS.The results indicated that CW had 14 new ingredients,such as 14-O-anisoylneoline and dehydro-mesaconitine,while N-demethyl-mesaconitine and aconitine disappeared.At the same time,it could significantly decrease the content of diester diterpenoid alkaloids and increase the contents of monoester diterpenoid alkaloids and amine-diterpenoid alkaloids.CH had 9 new ingredients from Hezi,like gallic acid,chebulic acid and shikimic acid.Neither the kinds nor the contents of compositions from Caowu in CH changed little.This suggested that the processing mechanism of CW reduced highly toxic components(diester diterpenoid alkaloids)and increased the content of lowly toxic components(monoester diterpenoid alkaloids and amine-diterpenoid alkaloids).Attenuated principle of CH may be related to the components of Hezi.In this experiment,the conclusion shows that the chemical constituents of CW and CH are essentially different,and the two methods have different toxicity reduction principles.