Quality Control Of Berberis Heteropoda Schrenk And Application Of Infrared Spectroscopy In Quality Control

Objective: To establish quality control methods of Xinjiang characteristic medicineBerberis heteropoda Schrenk, to study apply near infrared spectroscopy combined withspectra processing software and chemometrics methods to quantitative analysis andidentification of it. Method:1. With jatrorrhizine hydrochloride, palmatinehydrochloride and berberine hydrochloride as the reference substances.Different development system, different spotting sample volume, differentthin layer chromatography plate, different inspection, different temperatureand different humidity were investigated.2. To establish the method that withHPLC dual wavelength simultaneously determine the content of jatrorrhizinehydrochloride, berbamine hydrochloride, palmatine hydrochloride, and berberinehydrochloride in Berberis heteropoda. Determine and compare the content of4kinds ofalkaloids with different origin and different medicinal parts.3.10groups of Berberisheteropoda Schrenk were determined moisture and ash.4. To determine the totalflavonoids of Berberis heteropoda Schrenk by UV-Vis spectrophotometry,evaluate the antioxidant activity of the extracts from different parts ofBerberis heteropoda Schrenk in scavenging DPPH radicals,·OH radicalsand O2·-radicals.5. The various compositions in Berberis heteropoda Schrenkwere identified simultaneously with the mobile phases of microemulsion consisting withSDS/n-C4H9OH/n-C7H16/H2O. Deferent factors, such as the quantity of SDS, n-C4H9OH,and formic acid, which effect on the separation of the compositions were discussed.6. Todevelop rapid quantitative analysis method of berberine hydrochloride in Berberisheteropoda by NIR diffuse reflectance optic-fiber spectroscopy combine with PLS andPCR.7. To develop rapid origin and authenticity indentify of Berberis heteropoda by NIRdiffuse reflectance optic-fiber spectroscopy combine with PCA and Clustering analysis.Results:1. With jatrorrhizine hydrochloride, palmatine hydrochloride and berberinehydrochloride as the reference substances, TLC identification for Berberis heteropodaSchrenk was high specificity with toluene-ethyl acetate-methanol-isopropyl alcohol-ammonia water（6:3:2:1.5:0.5） as the developing system.2. Method of assayingalkaloids of Berberis heteropoda Schrenk by HPLC was established, and good linearitieswere obtained over the range of1.95⁶2.4μg·mL^-1（r=0.9998,n=6） for jatrorrhizinehydrochloride, the range of9.80³13.60μg·mL^-1（r=0.9997,n=6） for berbaminehydrochloride, the range of1³2μg·mL^-1（r=0.9999,n=6） for palmatine hydrochloride andthe range of4.93¹57.6μg·mL^-1（r=0.9999,n=6） for berberine hydrochloride, respectively.The average recoveris of jatrorrhizine hydrochloride, berbamine hydrochloride, palmatinehydrochloride and berberine hydrochloride were99.27%（RSD=2.17%,n=9）,98.46%（RSD=2.35%,n=9）,99.26%（RSD=2.94%,n=9）,98.46%（RSD=1.63%,n=9）,respectively.3. According to result of moisture and total ash determination, it is suggestthat moisture and total ash content of Berberis heteropoda Schrenk bark should be lessthan8.05%and7.82%, Berberis heteropoda Schrenk fruit should be less than15.48%and4.15%.4. Adopt ultraviolet spectroscopy to determine total flavonoids content of Berberisheteropoda Schrenk, it was high in fruit, then bark and root-bark; different officinal partsextracting solution of Berberis heteropoda Schrenk scavenging capacity of DPPH radicals results ranked in descending order as follows:fruit>bark>velamen, scavengingcapacity of·OH radicals results: root-bark> bark, scavenging capacity of O2·-radicalsresults: fruit>bark> root-bark.5. The berberine hydrochloride, jatrorrhizine hydrochlorideand palmatine hydrochloride in Berberis heteropoda were identified simultaneously withmobile phase of microemulsion form. The consist of microemulsion wasSDS:n-C4H9OH:n-C7H16:H2O=2.7:6.3:1.0:30.0（g/g）.6. NIR diffuse reflectance optic-fiberspectroscopy combine with PLSR and PCR method was used to create a quantitativemodel of calibration. It is shown that the root-mean-square error ofcross-validation（RMSECV） of the berberine hydrochloride are0.8102、1.0169, thecorrelation coefficients are0.9871、0.9795, the average recycle rate are101.40%、108.51%. Statistical test was used to analysis the dates and it’s shown that the differencebetween NIR prediction values and the values determinated by standard methods has nostatistical significance, it is indicated that NIR method can be used in rapid determinationof the contents of Berberis heteropoda Schrenk.7. PCA and Clustering analysis give abetter classification on39Berberis heteropoda Schrenk from3different geographicalorigins, the latter method was used to establish a discriminant analysis model which canbe used to identify the geographical origin of Berberis heteropoda Schrenk, by the model,14samples were analysed and the analysis results are basically identical with actual. It isindicated that NIR method is simple, accurate and can be used in rapid discrimination of Berberis heteropoda Schrenk from different geographical origins and authenticityidentification. Conclusion: To established quality control method of Berberis heteropodaSchrenk, it was providing a research foundation for the quality evaluation for alkaloidcomponents of Berberis heteropoda; Berberis heteropoda Schrenk extract has certainantioxidant capacity, this research can provide the basis for the development of naturalantioxidants; Microemulsion thin-layer chromatography as a new chromatographytechnology, it can be simple, accurate and efficient to identify the alkaloids of Berberisheteropoda Schrenk, and the amount of spotting is half of the ordinary chromatographicconditions, detection sensitivity was improved significantly, and could not use irritatingand toxic organic solvent such as stronger ammonia water, toluene, etc, reducedenvironmental pollution and the cost of experiments; Near infrared diffuse reflectionspectroscopy combined with chemometrics methods has the advantages including rapidanalysis, high efficiency, low cost, good reproductivity, pollution-free, withoutpretreatment and analysis of process does not destroy the sample etc. Such techniques canused to qualitative and quantitative analysis rapidly without destroy the sample, also canbe a auxiliary detection method for quality control of Xinjiang characteristic medicineBerberis heteropoda Schrenk.