A Preliminary Study On The Quality Control And Anti-inflammatory And Analgesic Effect Of Resina Populi

Objective:To preliminarily study the quality control method,chemical composition and analgesic effect of Resina Populi,and compare with the material composition characteristics of sap of Populus euphratica.Methods:（1）Combined with the literature,the morphology of the original plants and the properties of the medicinal materials were described.The calcium salt,sodium salt,potassium salt,iron salt,aluminum salt,magnesium salt,sulfate and chloride in Resina Populi were identified by the general identification test.The content of arsenic salt,heavy metal,extract,ash and weight loss on drying were examined.An ICP-MS method was established for the determination of Ca,Mg,Fe,Hg,CD and Pb in Resina Populi and sap of Populus euphratica.（2）The HPLC fingerprints of Resina Populi and sap of Populus euphratica were established by internal standard method.（3）A preliminary study on the chemical constituents of Resina Populi and its extracts by tube method.And UPLC-Q-TOF-MS was used to analyze the chemical constituents of the methanol extracts from Resina Populi.（4）Acute toxicity of Resina Populi and sap of Populus euphratica in mice were assessed.And evaluate the anti-inflammatory and analgesic effects of Resina Populi.Results:（1）Resina Populi are irregular lumps or crushed powder,and the surface is yellowish brown to brownish yellow.The smell is very slight,slightly bitter,astringent.Calcium salt,sodium salt,potassium salt,iron salt,aluminum salt,magnesium salt,sulfate and chloride were identified in 10batches of Resina Populi and 7 batches of sap of Populus euphratica.The content of arsenic salt and heavy metal is less than 2 mg·kg^-1and 20 mg·kg^-1respectively,the weight ratio of water-soluble extractives measured by cold leaching and hot leaching is not less than 62%and 64%,respectively.The weight ratio of ash is 57.76%～71.10%,the weight ratio of acid insoluble ash is 4.62%～8.21%,and the loss on drying is no more than 2%.There was a good linear relationship between the concentration of six elements and the response value in their respective concentration range（r>0.999 5）,the RSD（%）of the precision,stability and repeatability test results was less than 3.1%,the average recovery was 94.9%-100.4%,RSD was 1.5%-2.4%.The contents of Ca,Mg,Fe,Hg,Cd and Pb in Resina Populi are 5918.002±367.64 mg·kg^-1,5551.500±1574.29 mg·kg^-1,1652.213±566.24 mg·kg^-1,0.003±0.00 mg·kg^-1,0.030±0.02 mg·kg^-1,0.583±0.23 mg·kg^-1,respectively.The contents of Ca,Mg,Fe,Hg,CD and Pb in sap of Populus euphratica are539.262±62.96 mg·kg^-1,6799.027±166.48 mg·kg^-1,48.330±6.44 mg·kg^-1,0.002±0.00mg·kg^-1,0.008±0.00 mg·kg^-1,0.046±0.03 mg·kg^-1,respectively.（2）The conditions of HPLC fingerprint were as follows:the column was Agilent SB-C₁₈（250 mm×4.6 mm,5μm）,mobile phase was mixture of acetonitrile（B）and 0.1%formic acid solution（A）,gradient elution[0-60 min,12%-36%B;60-70 min,36%-38%B;70-130 min,38%B].The flow rate was 0.8 ml·min^-1;the column temperature was 40℃;the detection wavelength was[0-26 min,340 nm;26-70 min,300 nm;70-95 min,265 nm;95-130 min,300 nm],and the injection volume was 5μL.The 15 characteristic peaks in the fingerprint of Resina Populi were identified with the internal standard benzoic acid as the reference,the similarity of 10 batches of samples is more than 0.99,which has good quality consistency.11 characteristic peaks were identified in the fingerprint of sap of Populus euphratica,and the similarity of 7 batches of samples was greater than 0.99.（3）There are sugar,polysaccharide,saponin,tannin,phenols and alkaloids in resina of Resina Populi and its extracts.According to UPLC-Q-TOF-MS data,the chemical structures of 18components were analyzed in the positive ion mode,and the chemical structures of 12components were analyzed in the negative ion mode.（4）LD₅₀of Resina Populi is 7.06g·kg^-1(95%CI 6.63～7.51 g·kg^-1),LD₅₀of sap of Populus euphratica is 7.75g·kg^-1(95%CI7.02～8.57 g·kg^-1),the acute toxic symptoms of mice include decreased activity,inactivity,unstable standing and uncoordinated movement.The results of pathological examination of the dead mice showed that the high-dose herbal medicine had certain toxicity to the gastrointestinal tissues of mice.Compared with the control group,the analgesic effect of different doses of Resina Populi on mice was significant,and there was a dose-dependent relationship.Conclusion:There are differences in the material basis between Resina Populi and sap of Populus euphratica.The results of this study provide a reference for the improvement of the quality standard of Resina Populi.The acute toxicity of Resina Populi to mice was similar to sap of Populus euphratica.Resina Populi had definite analgesic effect on mice.