Study On The Improvement Of Quality Standard Of Temuren-5 Powder

Objective: To improve the quality evaluation system of Temuren-5 powder and lay an experimental foundation for upgrading the quality standards.Methods:(1)The components of berberine hydrochloride,gallic acid and geniposide in Temuren-5 powder were identified by TLC.(2)The properties,appearance uniformity,moisture contents and total ash contents of 10 batches of Temuren-5 powder were inspected.(3)The pretreatment process of Temren-5 sample samples was optimized according to the Qb D concept.(4)For 10 batches of Temune-5 powder,the contents of gallic acid,gardenoside,methyl gallate,corilagin,crocin I,genistein,granin β-1,2,3,4,6-pentagallic acid glucose were determined.(5)The fingerprint of Temuren-5 powder was established,and the similarity of the fingerprint was evaluated.Compare with the control sample and identify the common peaks were identified by comparing with chromatogram of reference substance.SPSS 26.0 software was used to carry out systematic cluster analysis,principal component analysis and partial least squares discriminant analysis on common peaks.Results:(1)The specificity of TLC method for the identification of gallic acid,gardenoside and berberine hydrochloride in Temuren-5 powder is good.In the test sample chromatography,the same color spots are displayed in the corresponding position of the control medicinal material chromatography..(2)Color appearance of Temuren-5 powder is uniform.Water content and total ash content range from 3.81 to 4.41% and 26.20% to30.59%,respectively.(3)The parameters of optimized pre-treatment process are as follows:weigh about 1.0g of Temuren-5 powder,put it into a 10 m L brown volumetric flask,add70% methanol,ultrasound extract at 30℃ for 15min(100W,40 k Hz),cool it,dilute to volume with 70% methanol solution,shake it well,stand still,accurately measure 2m L solution into a 25 m L brown volumetric flask,dilute it to volume,shake well,and the supernatant was filtrated by 0.22 μm microporous membrane,take the continuous filtrate and get it.(4)Within 0.009～0.298mg/m L,0.026～0.418mg/m L,0.001～0.036mg/m L,0.033～1.051mg/m L,0.004～0.118mg/m L,0.008～0.252mg/m L,0.014～0.452mg/m L,0.007～0.236mg/m L,the mass concentration of gallic acid,pomegranate glycoside,methyl gallate,gardenoside,corilagin,gentianoside,β-1,2,3,4,6-pentapacyl glucose,crocin I were linear with the chromatographic peak area.The average sample recovery rate of the above 8 substances was 103.2%,104.0%,93.2%,93.9%,93.7%,105.1%,98.7% and98.6%,respectively.The RSD for the precision,stability and repeatability test met the methodology requirements.The content of gallic acid is 3.010 ± 0.134(mg/g),the content of pomegranate is 9.752 ± 0.431(mg/g),the content of methyl gallate is 0.649 ± 0.124(mg/g),the content of genistein is 4.360 ± 0.677(mg/g),the content of gardenoside is13.046 ± 1.096(mg/g),and the content of corilagin is 1.522 ± 0.063(mg/g),the content ofβ-1,2,3,4,6-pentagalloyl glucose was 7.490 ± 0.280(mg/g),and the content of crocin I was 3.511 ± 0.471(mg/g).(5)A total of 23 common peaks were calibrated from the fingerprint of 10 batches of Temuren-5 powder,the similarity ranged from 0.979 to0.903.By comparison with the retention time of the reference substance and the UV spectrum,common peak 2 is gallic acid,and peak 5 is pomegranate glycoside α,peak 7 is methyl gallate,and peak 8 is pomegranate β,peak 9 is gentianoside,peak 11 is gardenoside,peak 13 is corilagin,peak 19 is ellagic acid,and peak 21 is β-1,2,3,4,6-pentagalloyl glucose,peak 22 is crocin I.Among the 10 batches of samples,S2,S8 and S10 belong to Class I,S5,S7 and S9 belong to Class Ⅱ,and the rest belong to Class Ⅲ.The cumulative variance contribution rate of the first four principal components is 91.62%.Peak 18,peak 19(ellagic acid),peak 20,and peak 23 are four chromatographic peaks of different markers.Conclusion: The systematic study on the identification,inspection,content determination and fingerprint of Temune-5 powder is helpful to comprehensively improve the quality standard of Temune-5 powder.The results of this paper can provide experimental data for its quality control,rational and safe drug use,and further development and utilization.