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Objective: To investigate the chemical constituents from the stems of Wisteria sinensis. Methods: The 14 chemical constituents were isolated and purified by silica gel, polyamide, Sephadex LH-20 column chromatography, and semi-preparative HPLC, and their chemical structures were identified by physico-chemical constants and spectral data. Results: Fourteen isoflavones were isolated from the methanol extract of the stems of W. sinensis and identified as tectorigenin-7-O-β-D-(6″-O-acetyl)-glucoside (1), 7,3',4'-trihydroxyisoflavanone (2), formononetin (3), afromosin (4), prunetin (5), biochanin A (6), gliricidin (7), 8-O-methylreyusin (8), tectoridin (9), genistin (10), sissotrin (11), ononin (12), kakkanin (13), and kushenol O (14). Conclusion: Compound 1 is a new compound named as tectoridin A, and compounds 2, 5, 7, 9-11, 13, and 14 are obtained from the genus Wisteria for the first time.
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Objective:To optimize the water extraction and alcohol precipitation technology of Xuanfei Zhike granule .Methods:Orthogonal test was used to investigate the effects of adding water , decocting time and boiling time on the water extraction , and the effects of relative density , alcohol precipitation concentration and alcohol precipitation time on the alcohol precipitation technology by taking comprehensive score including the amount of hesperidin , the amount of tectoridin and the yield of dry cream as the indices .Re-sults:The preferred water extraction technology was as follows: added 10 times water and extracted 1.5 h firstly, and then added 8 times water and extracted twice with 0.5 h for each.The preferred alcohol precipitation technology was as follows:concentrated the wa-ter extraction to a relative density of 1.05 (measured at 60℃), slowly added 95％ethanol to 80％alcohol solution and stored 18 h at low temperature .Conclusion:The optimal water extraction and alcohol precipitation technology is stable and feasible , which can pro-vide reference for the standardized production of Xuanfei Zhike granule .
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Objective To establish a quality standard for compound Heishen granules .Methods Scrophulariae Radix and Belamcandae Rhizoma were identified by TLC .HPLC was used to determine the content of cinnamic acid ,tectoridin and irisflo-rentin .The HPLC was performed on a column of Kromasil-C18 (150 mm × 4 .6 mm ,5 μm)with a mobile phase of acetonitrile (A)-0 .1％ hydrochloric acid (B)at a temperature 30 ℃ .The detection wavelength was set at 270 nm and the flow rate at 1 ml/min .Results The TLC method had good specificity without interference from negative control .The calibration curve showed a good linear relationship within the range of 16 .22-113 .57μg/ml for cinnamic acid(r=0 .9998) ,48 .19-337 .34μg /ml for tectoridin(r= 0.9998)and 16.40-114.80 μg/ml for irisflorentin(r= 0.9999) .The average recoveries were 99.23％ , 98.82％ ,99.17％ .Conclusion The established method is rapid ,accurate and reproducible .It can be used in the quality control of compound Heishen granules .
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OBJECTIVE:To establish a method for the decoction of isoflavone content and its acid hydrolysis conversion rate in the flower of Pueraria lobata. METHODS:Using the flowers of Pueraria lobata as raw material,the isoflavone with main com-ponent of tectoridin in the flower of P. lobata was prepared with ethanol,ethyl acetate extracted,ethanol recrystallized and puri-fied,and it was converted to tectorigenin with hydrolysis in hydrochloric acid. By screening the solvent and wavelength,UV spec-trophotometry was established to determine tectovidin and tectorigenin,and calculate the isoflavone content and acid hydrolysis con-version rate of tectoridin(expressed by the relative percentage of tectorigenin). It was compared with HPLC detection results,the accuracy of UV method was evaluated. RESULTS:The solvent was 70％ethanol solution containing 1％triethylamine,and the iso-flavone content was detected at wavelength of 339,274 nm. The linear range of tectoridin was 8.80-29.33 nmol/mL(r=0.9999). RSDs of precision(n=6),stability(n=5)and reproducibility(n=6)tests were lower than 1.94％;average recovery was 99.7％(RSD=1.77％,n=9). There were no statistical significances in the contents of total flavonoids (UV:17.64-25.55 nmol/mL vs. HPLC:17.39-24.40 nmol/mL) and the relative percentage of tectorigenin (UV:57.65％-87.59％ vs. HPLC:55.62％-91.14％). CONCLUSIONS:The established method is accurate,reliable,and can be used for the rapid determination of acid hydrolysis con-version rate of tectoridin.
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OBJECTIVE:To optimize and improve the quality standards of Daziran wuhua yin. METHODS:TLC was used for the qualitative identification of Cichorii Herba,Glycyrrhizae Radix et Rhizoma and Gossampinus malabarica. HPLC was used for the contents determination of chlorogenic acid and tectoridin. The column was Diamonsil C18 with mobile phase of acetoni-trile-0.4% phosphoric acid(pH adjusted with to 2.05 with triethylamine)(10∶90,V/V,chlorogenic acid or 18∶82,V/V,tectoridin) at a flow rate of 1 ml/min,the detection wavelength was 327 nm(chlorogenic acid)or 265 nm(tectoridin),the column tempera-ture was 25 ℃,and the injection volume was 10 μl. RESULTS:TLC of Cichorii Herba,Glycyrrhizae Radix et Rhizoma and G. malabarica showed clear spots and good separation. The linear range was 19.7-98.7 μg/ml for chlorogenic acid (r=0.999 5) and 25.4-126.8 μg/ml for tectoridin(r=0.999 8);RSDs of precision,stability and reprosucibility tests were lower than 4%,average re-coveries were 97.01%-104.98%(RSD=3.64%,n=9)、98.44%-104.58%(RSD=1.84%,n=9). CONCLUSIONS:The optimized and improved standard is helpful for the quality control of Daziran wuhua yin.
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Objective To optimize the extraction technology of flos Puerariae lobata isoflavone .Methods The flos Pu-erariae lobata isoflavone was distilled by ethanol circumfluence .Total flavonoids ,tectoridin and tectorigenin extracted from Puerariae using the UV and HPLC spectromertry methods were taken as evaluation indexes .Extraction technology was opti-mized with L9 (34 ) orthogonal test on the base of single observation of ethanol concentration ,solvent dosage and distilling time . Results The best extraction technology of flos Puerariae lobata isoflavone was :to add 12 times the amount of 70% ethanol for 90 minutes for the first time ,and 10 times the amount of 70% ethanol for 60 minutes for the second time .Conclusion The op-timized extraction process of flos Puerariae lobata isoflavone is reasonable and feasible ,and it can offer reference to actual pro-duction .
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This study was aimed to establish the quality and quantity analysis methods of tectoridin contented in the Iris tectorum Maxim. Thin layer chromatography (TLC) method was used to give quality analysis on tectoridin in I. tectorum. And the HPLC method was used to give quantity analysis on tectoridin in I. tectorum. The Diamonsil C18 (200 mm í 4.6 mm, 5 μm) column was used as analytical column. The acetonitrile-0.2% phosphoric acid (20:80) was used as mobile phase at the flow rate of 1.0 mL·min-1. The detection wavelength was 265 nm and the column temperature was 25℃. The results showed that the quality analysis of I. Tectorum had specific identification with-out the interference of other ingredients. The amount of inlet tectoridin had a good linearity with the response val-ue of peak area in the range of 0.12~2.22 μg. The average recovery rate was 100.96%. It was concluded that this method was simple and reproducible, which can be used in the quality control of I. tectorum.
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Objective:To establish an HPLC method for the determination of four kinds of isoflavone compounds including daid-zin, tectoridin, daidzein and tectorigenin in the extracts of Pueraria lobata flowers. Methods:The separation was performed on an Agi-lent C18 column(250 mm × 4. 6 mm, 5 μm) using a mobile phase of methanol-acetonitrile-water(2∶1∶2). The flow rate was 1. 0 ml· min-1 ,and the detection wavelength was 264nm. The column temperature was 25℃ and the sample size was 20 μl. Results:The de-tection could be accomplished within 10 minutes with good separation and specificity of four isoflavone compounds with the retention timeof3.4,3.8,5.7and7.2min,respectively. Thelinearrangewas0.784-78.440 μg·ml-1(daidzin),2.000-200.000 μg· ml-1(tectoridin) and 0.800-80.020 μg·ml-1 (daidzein and tectorigenin),and the relative coefficient was 0.999 9, 0.999 8, 0. 999 7 and 0. 999 9, respectively. The average recovery was 100. 54%(RSD=1. 66%,n=6),100. 03%(RSD=1. 00%, n=6), 99. 48%(RSD=1. 76%, n=6) and 100. 92%(RSD=2. 26%, n=6), respectively. Conclusion: The method is simple, rapid and accurate with good repeatability, which can be used in the rapid determination of isoflavone compounds in the flowers of Pueraria loba-ta.
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Objective To study the non-covalent binding between the tectoridin(TE) and plasma proteins.Methods The molecular weights of TE,human serum albumin(HSA),?1-acid glycoprotein(AAG) and the protein-drug complexes were measured by the electrospray ion trap mass spectrometry(ESI-MS).The maximum stoichiometric ratios were obtained according to the molecular weight change of the complexes before and after binding reaction.The binding constants(K) of the complexes were calculated by the Scatchard equation.The main sorts of binding force of the complexes were deduced according to the relationship between the reaction temperature and the thermodynamic parameters(?H and ?S).Results The K of the complexes were 1.914?104 mol/L for TE-HSA and 5.893?104 mol/L for TE-AAG,and the number of binding sites were 7.8 and 3.3,respectively.The main sorts of binding force between TE-HSA or TE-AAG were static-electricity gravitation.Conclusion ESI-MS is a good method for studying of the TE-protein non-covalent binding with some advantages in sensitivity,high-speed,and accuracy.
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OBJECTIVE:To determine the content of Tectoridin in Rhizoma Iridis Tectori by HPLC. METHODS: The chromatographic separation was performed on Diamond ODS C18 column(150 mm?4.6 mm,5 ?m) with mobile phase consisted of MeOH-0.5% acetic acid(3∶7) at a flow rate of 0.8 mL?min-1. The detection wavelength was set at 265 nm.RESULTS: Good linear relationship was achieved over the concentration range of 0.079 2～0.396 0 ?g for tectoridin with an average recovery of 99.80%(RSD=1.09%,n=6). CONCLUSION: The method is sensitive, simple and accurate, and it can be used for the quality control of Rhizoma Iridis Tectori.