Study On The Substance Benchmarks Of Classical Prescription Mahuang Decoction

Mahuang Decoction is a classic antiseptic prescription.It was first recorded in Shanghanlun compiled by Zhang Zhongjing in the Han Dynasty.It is composed of Ephedra Herba(Mahuang,MH),Cassia Twig(Guizhi,GZ),stir-baked Armeniacae Amarum Semen(Chankuxingren,CKXR)and honey-roasted licorice(Zhigancao,ZGC).It has the effects of inducing sweat and dispelling exogenous evils,and ventilating lung and relieving asthma,and indications for exogenous wind-cold manifestations,aversion to cold and fever,head and body pain,panting without sweating,thin white tongue coating,and floating pulse.It is commonly used clinically to treat colds,influenza,acute bronchitis,bronchial asthma,etc.In a bid to promote inheritance and innovation of traditional Chinese medicine industry,the "Catalogue of Ancient Classic Prescriptions(First Batch)" released by the State Administration of Traditional Chinese Medicine in 2018 included 100 classic prescriptions such as Mahuang Decoction.In this dissertation,Mahuang decoction was investigated from the following four aspects:the prescription history and herbal textual research,substance benchmarks preparation precudures,substance benchmarks quality control and chemical composition characterization of Mahuang decoction.By consulting the photocopy of ancient books,research documents and modern books,etc.,the prescription and historical evolution of Mahuang Decoction were studied,and the prescription usage,dosage and function were clearly identified as:MH 41.4 g,cassia twig 27.6 g,bitter almond 23 g,and roasted licorice 13.8 g,decoct MH with 1800 ml of water to 1400 ml,add cassia twig,bitter almonds and roasted licorice,boil to 500 ml,and administered warm in three times.According to historical records and research documents,the medicinal composition of Mahuang Decoction was verified,and its medicinal plant sources are confirmed to be Ephedra sinica Stapf.,Cinnamomum cassia Presl.,Prunus armeniaca L.var.ansu Maxim.and Glycyrrhiza uralensis Fisch.After textual research,bitter almond is found as a multi-sourced herbal medicine,which is consistent with the records in the "Chinese Pharmacopoeia".Taking the market and origin factors into account,mountain apricot is selected as the medicinal origin of bitter almond.Preparation procedure was investigated by the principle of "inheriting classics,keeping integrity and innovation",and the original record in "Treatise on Febrile Diseases"and "Notice on Printing and Distributing the Management Regulations of Traditional Chinese Medicine Decoction Chambers in Medical Institutions".Single factor investigations of soaking time,decoction time,first decoction time,added water and decoction times were conducted to optimize the preparation tecnology by comparing with reference to ancient decoction,and the indicatiors of extraction rate,content,fingerprint spectrum and transfer rates.Finally,the optimal preparation procedure was determined as follows:weigh 45 g ofMH.30 g of GZ,25 g of CKXR and 15 g ZGC in a 3500 ml electric casserole,and add 450 ml(10 times)and 700 Soak MH and other three flavor decoction pieces in 700 ml(10 times)water for 30 min.Fry MH first,heat to boiling,simmer for 10 minutes,add cassia twig,bitter almonds,roasted licorice pieces and their soaking liquid,heat to boiling,simmer for 1 hour,and filter.Then add 920 ml(8 times)water to the dregs,heat to boiling,and simmer for 0.5h,filtered.The filtrates were combined and lyophilized directly.Since cinnamaldehyde in cassia twig was a volatile component in Mahuang Decoction,this component was lost duing the evaporation.Therefore,it was selected to freezedry directly to fully reflect the chemical substance basis of Mahuang Decoction.The research on the quality control method of Mahuang Decoction’s substance benchmarks mainly included three aspects:thin-layer chromatography identification,fingerprint spectrum and content determination.MH,GZ,CKXR and ZGC were identified in three thin-layer chromatography conditions established in this study.Methodological verification showed that each method had good specificity and durability.Using ephedrine hydrochloride,pseudoephedrine hydrochloride,amygdalin,glycyrrhizin,cinnamic acid cinnamic aldehyde and glycyrrhizic acid as the reference peaks,the preparations of the test product were investigated and the chromatographic conditions were optimized to establish the fingerprinting method of Mahuang Decoction.The method verification results show that the method has good precision,stability and specificity.The four chemical components of ephedrine hydrochloride,pseudoephedrine hydrochloride,amygdalin and glycyrrhizic acid were used as the quantitative determination markers of Mahuang decoction.Three quantification methods were established and validated through optimization of the preparation and chromatographic conditions.Through UPLC-LTQ-Orbitrap-MS combined with molecular network data processing method,the different types of chemical components in Mahuang Decoction can be quickly classified.In the negative ion mode,the chemical components are aggregated into 20 molecular networks,mainly involving flavanone-O-glycosides.Triterpene saponins,flavonoids-Cglycosides,proanthocyanidins,phenolic acids and glycosides,etc.,according to the classification results,combined with the self-built Mahuangtang compound database,the chemical composition of Mahuangtang was characterized,and a total of 156 compounds were identified.In this study,the substance benchmarks of Mahuang Decoction,including the establishment of the quality control method of Mahuang Decoction and clarification of the chemical substance basis of substance benchmarks were systematically investigated,which could provide a quality evaluation method for the later development of the classic Mahuang Decoction preparations,and provide a scientific basis for the safety and efficacy of Mahuang Decotion.