Study On The Evaluation Of Mongolian Medicine Topical Ulcer And Realgar Quality

Objective:Ethno medicine is the cultural treasure of traditional Chinese medicine,and it is widely used.Mineral medicine is an important part of ethno medicine,but the detection methods are relatively backward,and lacking of perfect quality standards.External ulcer powder is the common medicine of gynecological diseases and skin diseases.External ulcer powder are consist of gypsum rubrum(cold processing),realgar,cinnabar,vermilion,concha haliotidis(calcined),and musk.Currently there is no perfect quality standard for external ulcer powder,especially for the evaluation of heavy metal and harmful element.Therefore,it is important to evaluate the quality of external ulcer powder.The research used traditional and modern technology to analyze and evaluate the external ulcer powder,and to provide a reference of quality control and clinical safety.To provide reference for the quality research of ethnic medicine,especially mineral drugs,in order to improve the evaluation method of national medicine.In the meanwhile,the research used zebrafish to analyze and evaluate the toxicity of realgar.A new way explored evaluates the toxicity of mineral medicines.Methods:1.Firstly,using polarized microscopic to analyze the external ulcer powder of different batches and the realgar of different producing areas.The composition of external ulcer powder and the associated mineral of realgar were preliminarily identified.Further analysis was made by combining Raman spectroscopy and X-ray diffraction to identify the components of exultant ulcer powder,and identified the types of gypsum rubrum and the impurities of realgar.The combination of three methods can conduct quick and accurate qualitative identification of external ulcer powder and realgar.2,The external ulcer powder and realgar were pretreated,and then the inductively coupled plasma mass spectrometry was used to measure.There were 38 kinds of inorganic elements to be tested,including Li,Be,Na,Mg,P,K,Ca,Sc,Ti,V,Cr,Mn,Fe,Co,Ni,Cu,Zn,Ga,Rb,Sr,Nb,Mo,Ag,Cd,In,Sn,Cs,Ba,Ta,W,Tl,Pb,Bi,Th,U,As,Sb and Hg.Inorganic elements fingerprints were established by construct content distribution curves of external ulcer powder and realgar.Then using analysis software SPSS 20.0 to conduct principal component analyses,and screening out the principal inorganic elements.Analyzing external ulcer disperses and realgar by comparing the relevant existing standards of harmful elements and heavy metal elements.3.To evaluate the safety of realgar by using model organism zebrafish.The realgar can be processed into micron-sized superfine powder,and the suitable dispersant and dispersion conditions are selected to make stable realgar suspension.Using 72 hpf zebrafish as a model to evaluate cardiotoxicity,hepatotoxicity,neurovirulence,nephrotoxicity,and gastrointestinal irritation of realgar.The research was established by methods of toxicity evaluation of traditional Chinese medicine in zebrafish juvenile fish.Verifing the accuracy and feasibility of toxicity assessment by using zebrafish model in associate with traditional animal acute toxicity tests,reports and clinical researchers.Results:1.Guizhou realgar containded more impurities.And cinnabar,realgar,gypsum rubrum were identified in external ulcer powder by using polarized light microscope.In external ulcer powder,gypsum rubrum was red gypsum but not calcite.The polarized light microscope was suitable for preliminary discrimination of medical material and preparation.Types and structures of compounds were analyzed by using the fingerprint which was made by Raman spectrum and x-ray diffraction method.By using Raman spectrum and x-ray diffraction method,we found out quartz of Guizhou realgar,and there was As2O5 in Hunan realgar,Hubei Realgar,Sichuan realgar,and Guizhou realgar.Calcium sulphate dihydrate was detected out in two batches of eight batches external ulcer disperses.Other powders only had Calcium sulfate.Crystal water in red gypsum may be affected by process.2.Elemental mercury content was the most,calcium content was the second,and arsenic element content was more than 10 mg·g-1 among the 38 kinds of inorganic elements that were tested by ICP-MS in external ulcer powder.Elemental elements,calcium,and arsenic may mostly come from cinnabar,vermilion,gypsum rubrum,and realgar.Pearson's coefficients were all above 0.99 for the inorganic elements contents in eight bathes of external ulcer powder.Inorganic elements fingerprints of external ulcer powder were established.The principal elements of external ulcer powder were K,Rb,Dc,U,Zn,Mn,Hg,Ti,Nb,Cs,Sc,Ga,P and Na by using principal component analysis.These elements can be used as key elements in the quality evaluation of external ulcer powder.In addition to As,the contents of Sb,Fe,Na,Mg,Sr and P were higher,and these elements were related to the deposit deposition of realgar.The amount of Pb and Hg in some of batches of external ulcer powder,and Cd of realgar was more than some standard limit.The quality control should be strengthened.3.The average particle size of ultramicro realgar was about 500 nm by using low temperature high speed ball mill,and stable disperse system can be made by choosing PVP as dispersant.Mass ratio of realgar compare PVP was one compare three.The duration of ultrasound was 20 min.The temperature of ultrasound was 60 Celsius degree.Using the realgar stable disperse system into zebrafish model,the toxicity results showed that realgar may have hepatotoxicity and gastrointestinal irritation in zebrafish larvae.The hepatic region of zebrafish showed dark color and yolk sac absorption delayed.The other toxicities were not obvious.Acute toxicity observation based on mice,and it showed that realgar may have hepatotoxicity and nephrotoxicity.Other toxicities were not obvious.The experimental period of toxicity test in zebrafish is shorter than in mice,and much more samples.Most of toxicity index can be observed.Zebrafish model has more advantages compare to traditional animal model.Conclusion:The research established qualitative and quantitative analysis method for external ulcer powder by combining polarization microscopic identification,Raman spectrum,x-ray diffraction technique,and ICP-MS,and provided reference for identification and quality evaluation of external ulcer powder.The toxicity results of realgar in zebrafish model compared to traditional animal model are highly consistent.It means zebrafish can be used as an animal model to evaluate realgar toxicity.The research provides new reference for mineral medicine quality evaluation.