Research On Quality Group Standards And Evaluation Of Radix Isatidis In Heilongjiang Province

Radix isatidis is the dried root of Isatis indigotica Fort,a cruciferous plant.Indications for exogenous fever,sore throat,warm toxin rash.It is often used clinically to treat influenza and has high medical value.At present,the quality of Radix isatidis is uneven,and there is a lack of an evaluation system for grading its quality.The current quality standard content measurement items of Radix isatidis only use(R,S)-Goitrin as the detection index,and it is difficult to standardize the overall quality of Radix isatidis.Therefore,It is necessary to establish the basis for the quality classification of Radix isatidis and to study the quality control method of multiindicators.In this study,a total of 38 batches of Radix isatidis medicinal materials mainly in Heilongjiang Province were collected.According to the 2020 edition of "Chinese Pharmacopoeia",they carried out character identification,microscopic identification,thin layer identification,moisture detection,total ash and acid insoluble ash detection,and extracts.(R,S)-Goitrin content determination,based on the current standards,38 batches of medicinal materials all meet the standards,and the content of(R,S)-Goitrin medicinal materials in Heilongjiang Province is about 1.5 times that of other provinces,of which The average content of Daqing City and Qiqihar Tailai County is the highest,respectively 0.118%and 0.123%,and the content limit of(R,S)-Goitrin is raised to 0.025%when formulating group standards.Quantify the above-mentioned test results and carry out cluster analysis combined with SPSS software to obtain the group quality standards and quality grading evaluation basis of Radix isatidis,and draw up equal weights for each test item.32 batches of Radix isatidis from Heilongjiang Province were scored and graded,and the results It shows that the comprehensive quality of Radix isatidis in Daqing is the best,moreover,the overall quality of Radix isatidis in Heilongjiang Province is slightly better than other major producing areas.GC-MS and UPLC-MS were used to detect the organic pesticide residues in the collected Radix isatidis,and no residues of banned pesticides were detected,indicating that the use of 33 banned organic pesticides did not occur during the growth of Radix isatidis.ICP-MS was used to detect heavy metals and harmful elements.Pb,Cd,As,Hg,Cu and other 6 metal elements Na,Ca,Mg,Cr,Ni,Ba in Radix isatidis,among which Pb,Cd,As,Hg,Cu The contents of Ca and Mg,which are beneficial to the human body,are in the range of 3198.02～9711.96mg/kg and 1174.03～3018.86mg/kg,which have a high degree of enrichment.such essential trace elements as Cr and Ni also exist in a small amount in Radix isatidis.The establishment of the multi-index content determination method inspected the detection wavelength,the choice of mobile phase,the column temperature,the flow rate,etc.secondly,the extraction of Radix isatidis was investigated,including the extraction solvent,extraction time,extraction method and other conditions.Finally,it was determined to extract the index components by ultrasonication of 5%methanol for 30min.The chromatographic conditions were column temperature:30℃;injection volume:10μl;flow rate:0.8mL/min;detection wavelength:254nm,230nm.Mobile phase:methanol-water solution.The content of 6 index components guanosine,adenosine,adenine,(R,S)-Goitrin,uridine,and Clemastanin B in the sample was determined by gradient elution dual-wavelength detection.The research found that Adenosine in the extract of Radix isatidis will be continuously degraded to generate adenine due to the action of endogenous components,and the ratio of the reflected substance is 1:1.guanosine,(R,S)-Goitrin,Clemastanin B,adenosine and adenine content are 210.71～844.45ug/g,105.85～450.44ug/g,962.32～3734.86ug/g,0～499.29ug/g,1.37～2.74μmol/g,among which the average content of Clemastanin B in Heilongjiang medicinal materials was higher than that in other provinces,and there were a batch of other provinces that did not detect this ingredient.Based on the multi-index content determination method,the fingerprints of 32 batches of Radix isatidis samples in Heilongjiang Province were established,and the feasibility of the method was investigated through precision experiments,repeatability experiments,and stability experiments.The established fingerprints had 11 common peaks.Six of the ingredients were identified,namely uridine,guanosine,adenine,(R,S)-Goitrin,adenosine,and Clemastanin B,and the similarity calculation results of 32 batches of medicinal materials were all above 0.944.The established fingerprints can better reflect the quality characteristics of Radix isatidis in Heilongjiang Province.Using DRS Origin software combined with double-standard linear calibration method(standard retention time method)to conduct qualitative analysis on 6 index components and establish the template of the double-standard linear calibration method for the 6 components,and screened 13 out of 20 chromatographic columns The chromatographic column suitable for this method is compared with the relative retention time method,and 4 chromatographic columns that have not been established are selected for analysis by two qualitative methods,and it is found that the double-standard linear correction method can predict and qualitatively analyze the retention time.more reliable in terms of accuracy.