Study On The Fingerprint Of Alcohol Extract Of Polygonati Odorati Rhizoma And Raman Spectra Of Two Homoisoflavanones

Polygonati Odorati Rhizoma is the dried rhizome of Polygonatum odoratum（Mill.）Druce.It is a medicine and food homologous substance,which has the effects of nourishing,moistening,replenishing and quenching thirst.Widely used in medicine,health food,cosmetics and other industries.The resources of Polygonati Odorati Rhizoma are vast.Because of different habitats,the ingredients are different,resulting in differences in efficacy.Modern pharmaceutical research shows that the four homoisoflavanones（Ⅰ,Ⅲ,Ⅳ,Ⅴ）in Polygonati Odorati Rhizoma have strong biological activities in anti-cancer,bacteriostatic,anti-oxidation and non-enzymatic glycosylation of proteins.However,in the 2015 edition of“Chinese Pharmacopoeia”,in the detection of active ingredients in Polygonati Odorati Rhizoma,only“total polysaccharide≥6.0%”and“total alcohol extract≥50.0%”are included,which cannot fully reflect its quality.Based on the above situation,this paper for the first time systematically established the qualitative and quantitative analysis methods of the four homoisoflavanones（Ⅰ,Ⅲ,Ⅳ,Ⅴ）in Polygonati Odorati Rhizoma and the fingerprints of the alcohol extracts of Polygonati Odorati Rhizoma.Main content and innovation:1.Based on secondary chromatography HPTLC,research and establishment of the identification method of four homoisoflavanones in Polygonati Odorati Rhizoma was used for comparison and analysis of 20 batches of Polygonati Odorati Rhizoma samples.2.For the first time,based on TLC separation,in-situ Raman spectroscopy of two main homoisoflavanones（Ⅲ,Ⅳ）spots in Polygonati Odorati Rhizoma was carried out and applied to the identification of three batches of Polygonati Odorati Rhizoma samples.For the first time,an in-situ enrichment method for TLC main spots toward the center was established,which effectively increased the concentration of TLC components to be measured,and provided a new method for improving the sensitivity of Raman spectroscopy in situ detection.3.The UPLC fingerprint of 33 batches of Polygonati Odorati Rhizoma alcohol extract was established,13 common peaks were calibrated,and 4 of them were identified as four homoisoflavanones（Ⅰ,Ⅲ,Ⅳ,Ⅴ）.There were thirty-three batches of Polygonati Odorati Rhizoma with 19 similarities above 0.90.The use of principal component analysis（PCA）,partial least squares discriminant analysis（PLS-DA）and independent sample t test combined with the normal distribution analysis of the VIP values of various variables,divides the Polygonati Odorati Rhizoma of different origins into 5 group,six main different components were found,including three homoisoflavanones（Ⅲ,Ⅳ,Ⅴ）components.4.An Agilent Acquity UPLC system,Kromasil C₁₈（100 mm×2.1 mm,1.8μm）column was used,with 0.1%formic acid aqueous solution（A）-acetonitrile（B）as the mobile phase,gradient elution,the detection wavelength is 297 nm.For the first time,UPLC analytical methods was established analytical methods for the simultaneous determination of four homoisoflavanones in Polygonati Odorati Rhizoma,including internal standard method,one-test multi-evaluation method and external standard method.Methodological verification of the three quantitative methods shows that the internal standard method is similar to the one-test multi-evaluation method and meets the requirements of the Chinese Pharmacopoeia for quantitative analysis of Chinese medicine.The internal standard method used in 33 batches of Polygonati Odorati Rhizoma.the test results showed that the average content ofⅠ,Ⅲ,Ⅳ,andⅤwere 5.754μg/g,35.60μg/g,27.48μg/g,15.74μg/g,and the total content was 84.58μg/g.The content of the same ingredient in Polygonati Odorati Rhizoma in different producing areas is obviously different,among which 12 batches are lower than the lower limit of the linear range and 3 batches are higher than the upper limit of the linear range.