Study On The Substance Benchmark Of The Famous Classic Formula Pipa Qingfeiyin

Objective:Taking the famous classical formula of Pipa Qingfeiyin as the research object,the formulas textual research and historical evolution were combed and studied.The Pipa Qingfeiyin substance benchmark preparation technology was determined,and then the substance bench-mark quality standard was established,so as to provided reference for the quality control and evaluation of the famous classical formula Pipa Qingfeiyin.Methods:（1）By consulting the literature and referring to the principles of textual research on the key information of ancient famous classical formula and the table of key information of ancient famous classical formula（7 formulas）,this paper combs the textual research,historical evolution and reseach status of the famous classical formula of Pipa Qingfeiyin.（2）Thin layer chromatography（TLC）was used to identify the flavor of Pipa Qingfeiyin.（3）The UPLC-DAD fingerprint of Pipa Qingfeiyin was established by ultra performance liquid chromatography（UPLC）-diode array detector（DAD）.The retention time and absorption wavelength of the corresp-onding peaks were identified by comparing with the UPLC-DAD of the reference substance,and the peaks were assigned by comparing with the UPLC-DAD of the negative samples.The contents of phellodendrine hydrochloride,chlorogenic acid,berberine hydrochloride,palmatine hydrochloride and ammonium glycyrrhizinate were determined by UPLC-DAD fingerprint of Pipa Qingfeiyin.（4）The UPLC-ELSD fingerprint of Pipa Qingfeiyin was established by ultra performance liquid chromatography（UPLC）-evaporative light scattering detector（ELSD）.The retention time of the corresponding peak was determined by comparing with the sample of single drug and the control sample UPLC-ELSD.（5）According to the records of yizongjinjian and the management standard of traditional Chinese medicine decocting room in medical institutions,the substance benchmark preparation method of Pipa Qingfeiyin was investigated to optimize its preparation process.（6）15 batches of substance benchmark of Pipa Qingfeiyin were prepared,and its properties,paste yield,moisture content,fingerprint and content of index components were studied,the quality control standard was established.Results:（1）The key information such as medicinal parts,processing specifications,dosage conversion,functional indications and clinical positioning of Pipa Qingfei drink were determined,and the related research of Pipa Qingfeiyin was reviewed.（2）Licorice TLC identification was good,negative without interference,spots clear,resolution was good,Coptis TLC identification,resolution is good,spots clear.The TLC identification of other drugs was negative,with interference or unclear spots.（3）The UPLC-DAD fingerprint of Pipa Qingfeiyin was established,and the methodology was well validated.Twenty three chromatographic peaks were marked,of which four peaks were from the leaves of Eriobotrya japonica,one was from the leaves of Eriobotrya japonica and Cortex mori,nine from Coptis chinensis,one from Phellodendron amurense,five from Coptis chinensis and Phellodendron amurense,and three from Glycyrrhiza uralensis.At the same time,by comparing the retention time and absorption wavelength curve of the corresponding peaks,a total of 11 characteristic peaks were identified.A UPLC-DAD method for the determination of phellodendrine hydrochloride,chlor-ogenic acid,berberine hydrochloride,palmatine hydrochloride and ammonium glycyrrhizinate was established and the methodology was well validated.（4）The UPLC-ELSD fingerprint of Pipa Qingfeiyin was established,and the methodology was well validated.Six chromatographic peaks were marked,all of which came from Ginseng flavor.By comparing the retention time of the corresponding peaks with the reference substance,five characteristic peaks were identified,which were ginsenoside Rg₁,ginsenoside Re,ginsenoside Rf,ginsenoside Rb₁and ginsenoside Rc,respectively.（5）After process optimization,the substance benchmark of Pipa Qingfeiyin preparation method was determined as follows:Ginseng 1.12g,honey Loquat leaf 7.46 g,Licorice 1.12 g,Coptis chinensis 3.73 g,Cortex mori 7.46 g,Cortex phellodendri 3.73 g were placed in a 2 L electric ceramic decoction pot,water 300 m L was added to determine the total mass before decoction,and then the decoction was boiled for about11 min with 400 W of Wu fire,and then the slight boiling was maintained for about 60 W with 200 W of gentle fire when the total weight was reduced by 110 g,stoped heat,passed through 400 mesh sieve while it was hot,and filtered to obtain 140 m L of Pipa Qingfeiyin.（6）The average paste yield and moisture content of 15 batches were14.23%and 8.87%,respectively.The similarity of the two fingerprints corresponding to 15 batches of substance benchmarks was greater than 0.9,and the difference of the content of index components was small.Conclusion:The formula research and historical evolution information of Pipa Qingfeiyin combed in this paper was reliabled.The optimized substance benchmark preparation process of Pipa Qingfeiyin was stable and feasible,and the material standard quality standard of Pipa Qingfeiyin substance benchmark was initially established,which could comprehensively and effectively control its quality,and provide reference for the rational development and research of the famous classical formula of Pipa Qingfeiyin.