Study On The High Performance Liquid Chromatographic Fingerprint Of ShouTaiwan

Purpose:Shou Taiwan is the basic Prescription of reinforcing Kidney and antiabortion. It was created by Zhang Xichun in Qing period. It has very effect for protecting growth of the fetus. In this study, It established the chromatographic fingerprint of water and ethanol soluble component of shoutaiwan by using HPLC for identifying and evaluating its quality. We analyse 10 batch of finished products and herbs with HPLC. We choose the best condition from different preparation method, solid-mobile phase, elution gradients, detect wave-length, etc. We use the "Chinese medicine Chromatographic Fingerprint Similarity Evaluation Software" to identify it eligible or not. Finally, we established the HPLC chromatographic fingerprint of Shoutaiwan succefully.Methods and results:Experiment One:Based on the constitude and active ingredient of Shou Taiwan, this study reviews four extracting methods.Results:Combine with the aggregate analysis of pesticide effect and atlas, water and ethanol soluble component of shoutaiwan is bast in number, size and shape of the peak, also the active ingredient of the Shou Taiwan is most perfect and content of the component is most. Hence the water and ethanol soluble component of shoutaiwan is the best way to extract.Experiment Two:Analyse the Shou Taiwan and semen cuscutae, Taxillus chinensis, Dipsacus asper Wall, this study roughly monitors the the chemical constituents by thin layer chromatography method, clears the main composition and tries to find chromatographic condition of HPLC, aiming at the structure characteristic and rationalized features of the active ingredient of the end product.Results:According the simulation production flow of Shou Taiwan and refer the solution of Shou Taiwan, semen cuscutae, Taxillus chinensis and Dipsacus asper Wall with Rutin, quercetin, Quercitrin, hyperin, kaempferol, Dipsacaceae saponinsⅥ, oleanolic acid, finding spot corresponding to a lot of control article in thin layer of chromatographic (TLC), it proves that Shou taiwan and medicinal materials contain the component of the control article. So according the above character to choose suitable liquid condition, this study provide material basis for fingerprint.Experiment Three:Analyse the Shou Taiwan solutions for testing with HPLC, compare the different ways to deal with samples. Checking different chromatogram system and gradient elution procedure, taking the peak to be many and resolution to be good, and guaranteed the main chemical conponents occurs in the standard fingerprint, then finally the best solution and procedure. We chose 210nm,230nm,325nm,355nm,256nm switching to 210nm, and define the detece wavelength as the information of Chromatographic Peak. The research compares the results in different environment, such as Dikma Platisil ODS 5μm 250×4.6mm ser#:5031458, UltimateTM XB-C185μm,4.6×250mm ser#:210802004, Agilent HC-C18 Analytical 4.6×250mm 5-Micron P. N.518905-902, then chooses the best one to be the chromatographic column in HPLC fingerprint. Comparing different column temperature (25℃,30℃,35℃) effects to the abruption of chromatographic peak to choose the best temperature of chromatographic column. Above all, based on the Shou Taiwan fingerprint, checking the precision, stability, repeatability and intermediate precision (including different date, different people, different instrument) to find the scientific and feasibility of the methods. The study analyses ten groups of end products to find out if the precision is ok.Results:In this study, we used acetonitrile(A) with a-(0.05mol/L) KH2PO4-H3PO4 buffer solution pH=3.0(B) gradient mobile system. The detecting wavelength was set at 220 nm and the flow rate was 1.0ml/min.36 characteristic peaks was selected.We have known that there are five types materials in the sample solution of Shoutaiwan.They are flavonoids, organic acids, triterpenoid saponins, iridoids and andrograpuolide. We identificatied the chromatographic peaks which the peak area is bigger than 3. It was detected 36 chromatographic peaks. The number of chromatographic peaks which was belong to semen cuscutae was eight, The number of chromatographic peaks which was belong to Taxillus chinensis was seven, The number of chromatographic peaks which was belong to Dipsacus asper Wall was twentyone, the twelfth chromatographic peak was mixing peak. there are seven coupounds:Asperosaponin, Rutin, Quercetin, Quercitrin Hyperoside, Kaempferol, Chlorogenic Acid. Chlorogenic Acid is organic acids. Rutin, Quercetin, Quercitrin Hyperoside, Kaempferol are flavonoids. Asperosaponin is triterpenoid saponins. Hyperoside and Kaempferol are in the herb of semen cuscutae. Rutin, Quercetin, Quercitrin are in the herb of Taxillus chinensis. Asperosaponin and Chlorogenic Acid are in the herb of Dipsacus asper Wall. It defined the composition of Shoutaiwan deeply. It also established the Chromatographic Fingerprint of HPLC of semen cuscutae, Taxillus chinensis, Dipsacus asper Wall. The similiarity degree of three herbs were more than 0.900.Conclusion:First, seven kinds of materials detected by the fingerprint of Shou taiwan estabilished by HPLC respectively are:Asperosaponin, Rutin, Quercetin, Quercitrin Hyperoside, Kaempferol, Chlorogenic Acid.Second, compared the fingerprint of Shou taiwan with the fingerprint of corresponding herbs, primarily identifies Hyperoside and Kaempferol as the main component of semen cuscutae, Rutin, Quercetin and Quercitrin as the main component of taxillus chinensis, Asperosaponin and Chlorogenic Acid as the main component of dipsacus asper WallThis method shows high precision and good repeatability. So it can be used as the means to assess the quality of Shoutaiwan.