Quality Control Of Traditional Chinese Medicine By Nonaqueous RP-HPLC

Traditional Chinese Medicine is a treasure for all thepeople in the world. As we know, a part of the components in itis fat-soluble, and many of these components have biologicactivities. To determine the content of these fat-solublecomponents, the traditional method is NP-HPLC, RP-HPLCwith the mobile phase involving water, GC, etc. Compared withRP-HPLC, NP-HPLC is more tedious, too expensive and thereis too many influence factors. So it is unsuitable for a regularanalysis method. GC is generally used for the analysis of thesample that can be volatilized at the work temperature. So GC isunsuitable for the analysis of the drugs with high boiling pointor unstable at heat. And the retention time of these contents islong by RP-HPLC with the mobile phase involving water.However nonaqueous RP-HPLC has many good characteristicsin separate low-pole components: larger the load capacity ofcolumn; shorter the analysis time; longer the chromatographycolumn life. In this paper we studied nonaqueous RP-HPLCused in separation low-pole components in Traditional Chineseherb Medicine, Traditional Chinese patent Medicine andbiologic samples. We complete the separation of Oleanalic acidin Fructus Ligustri lucidi, nuciferene in lotus leaves,anthraquinone in sanhuang tablets, Niuhuangjiedu Soft Capsules,Zhigan -ning Capsules and plasma, feces and urine of rats. Theresults showed that nonaqueous RP-HPLC is simple, rapid, widelinearity and reproducible. And it can be used for a regularanalysis in production to determine low-pole contents intraditional Chinese medicine, and solve the problem of analysisof fat-soluble components.PART 1(1) A rapid method to determine Oleanalic acid in FructusLigustri lucidiObjective: To develop a rapid method to determineOleanalic acid in Fructus Ligustri lucidi.Method: The separation was performed on an inertsilC8(4.6mm×250mm,um)column with a mobile phase composedof methanol-acetonitrile=20:80. The detection wavelength was210 nm and the flow rate was 1.0 ml/min. The columntemperature was maintained at 30℃.Results: Under this condition, the capacity factor ofOleanalic acid exceeded 2.1, the theroy plate number exceeded7000, and the peak of Oleanalic acid separated from the othercontains.Conclusion: Compare with the traditional method,nonaqueous RP-HPLC is simple, rapid method to determineOleanalic acid in Fructus Ligustri lucidi.-aqueous RP-HPLC(2) Determination of nuciferene in lotus leaves by nonaqueousreverse phase high performance liquid chromatographyObjective: To develop a rapid method to determinenuciferene in lotus leaves.Method: A Kromasil C18 column (250×4.6 mm, 5.0 μm)with a mobile phase composed of acetonitrile-triethylamine(99.9:0.1) was used. The detection wavelength was 270 nm andthe flow rate was 1.0 ml/min.Results: The linear range for nuciferene was 0.238~80.42μg/ml. The average recovery of nuciferene was 99.2%.Conclusion: Compare with the traditional method,nonaqueous RP-HPLC is simple, rapid, wide linear andreproducible.PART 2(1) Determination of rhein, emodin, chrysophanol and physcionin sanhuang tablets by nonaqueous reverse phase highperformance liquid chromatographyObjective: To determine the content of rhein, emodin,chrysophanol and physcion in Sanhuang tablets by NonaqueousRP-HPLC.Method: A Kromasil C18 column (250×4.6 mm, 5.0 μm)with a mobile phase composed of methanol-acetic acid (99.9:0.1)was used. The detection wavelength was 254 nm and the flowrate was 1.0 ml/min.Results: The linear range for emodin, rhein, chrysophanoland physcion was 0.395~31.6 μg/ml, 0.420~33.6 μg/ml,1.00~80.2 μg/ml, 0.525~42.0 μg/ml respectively, the averagerecoveries of rhein, emodin, chrysophanol and physcion were98.2%, 99.5%, 100.1%, 101.2%.Conclusion: Compare with the traditional method,nonaqueous RP-HPLC is simple, rapid, wide linear andreproducible.(2) Determination of Emodin, Chrysophanol and Physcion inNiuhuangjiedu Soft Capsules by Nonaqueous RP-HPLCObjective: To determine the emodin, chrysophanol andphyscion in Niuhuangjiedu soft capsules.Methods: Nonaqueous RP-HPLC was used to determinethe contents of emodin, chrysophanol and physcion inNiuhuangjiedu soft capsules. The separation was performed onKromasil C18 column with methanol-peracetic acid (99.9:0.1) asa mobile phase and the wavelength of UV detector was 254nm.Results: The emodin, chrysophanol and physcion sampleshowed a good linear relationship at the range of 0.420μg/ml~33.6μg/ml, 1.00μg/ml~80.2μg/ml, 0.525μg/ml~42.0μg/mlrespectively. The average recoveries of the added sample were99.6%, 100.2%, 100.0% respectively. The RSD were 1.0%,0.4%, 0.4% respectively.Conclusion: The method is simple, accurate and rapid. Itcan be used for content determination of emodin, chrysophanoland physcion in Niuhuangjiedu soft capsules.(3) Determination of Emodin and Physcion in ZhiganningCapsules by Nonaqueous Reverse-Phase High PerformanceLiquid ChromatigraphyObjective: To determine the content of emodin andphyscion in Zhiganning Capsules by Nonaqueous Reverse PhaseHigh Performance Liquid Chromatography.Method: Nonaqueous RP-HPLC was used. The separationwas performed on Kromasil C18 column (250×4.6 mm, 5.0 μm)with the mobile phase of cosisting of methanol-acetic acid(99.9:0.1). The flow rate was 1.0 ml?min-1 and the detectionwavelength was 254 nm.Results: The linear ranges for rhein and physcion were0.420~33.6 μg?ml-1 and 0.525~42.0 μg?ml-1 respectively. Theaverage recoveries of rhein were 99.6%, 98.8% and 100.2% withcorresponding RSD of 1.02%, 1.31% and 0.97%. The averagerecoveries of physcion were 98.7%, 100.7% and 101.4% withcorresponding RSD of 1.6%, 1.4% and 1.5%.Conclusion: It is simple, rapid, accurate and reproduciblewith RP-HPLC to detect rhein and physcion.PART 3(1) Studies of Pharmacokinetics of Emodin in RhizomaPolygontum Cuspidatum and its CompoundObjective: To study the difference of the pharmacokineticsof emodin in Zhiganning Capsules and Rhizoma polygontumCuspidatum by Nonaqueous Reverse Phase High PerformanceLiquid Chromatography.Method: The rats were oral administered with the RhizomaPolygontum Cuspidatum extraction and Zhiganning capsules.After hydrolyze and extraction, the content of emodin in theplasma was determined by Nonaqueous RP-HPLC.Results: The concentration-time profiles of emodin fittwo-compartment model. The pharmacokinetics parameters,t1/2 ,AUC(0 α～∞),CL(s) and Cmax of emodin in RhizomaPolygontum Cuspidatum group and its componds group showedsignificant differences.Conclusion: The study shows that the two groups havesignificant differences in pharmacokinetics of emodin.