Clinopodium chinense, a traditional folk medicinal herb, has been used to treat abnormal uterine bleeding(AUB) for many years. Saponins and flavonoids are the main active components in C. chinense. To study the pharmacokine-tics of multiple components from the total extract of C. chinense(TEC), we established a sensitive and rapid method of ultra-perfor-mance liquid chromatography coupled with tandem mass spectrometry(UPLC-MS/MS) for simultaneous determination of five compounds in the plasma of AUB rats. After validation, the AUB model was established with SD female rats which got pregnant on the same day by gavage with mifepristone(12.4 mg·kg~(-1)) and misoprostol(130 μg·kg~(-1)). The established method was applied to the detection of hesperidin, naringenin, apigenin, saikosaponin a, and buddlejasaponin Ⅳb in AUB rats after the administration of TEC. The pharmacokinetic parameters were calculated by DAS 2.0. The five compounds showed good linear relationship within the detection range. The specificity, accuracy, precision, recovery, matrix effect, and stability of the method all matched the requirements of biolo-gical sample detection. The above 5 compounds were detected in the plasma of AUB rats after the administration of TEC. The C_(max) va-lues of hesperidin, naringenin, apigenin, saikosaponin a, and clinoposide A were 701.6, 429.5, 860.7, 75.1, and 304.1 ng·mL~(-1), respectively. All the compounds owned short half-life and quick elimination rate in vivo, and the large apparent volume of distribution indicated that they were widely distributed in tissues. Being rapid, accurate, and sensitive, this method is suitable for the pharmacokinetic study of extracts of Chinese herbal medicines and provides a reference for the study of pharmacodynamic material basis of C. chinense in treating AUB.
Administration, Oral ; Animals ; Apigenin/analysis* ; Chromatography, High Pressure Liquid/methods* ; Chromatography, Liquid ; Drugs, Chinese Herbal/chemistry* ; Female ; Flavonoids/analysis* ; Hesperidin ; Lamiaceae ; Mifepristone ; Misoprostol ; Oleanolic Acid/analogs & derivatives* ; Plant Extracts/chemistry* ; Rats ; Saponins ; Tandem Mass Spectrometry/methods* ; Uterine Hemorrhage

This study explored the correlation between color and chemical components of Chrysanthemi Indici Flos(CIF), aiming at providing a reference for its procurement, evaluation, and breeding. Colorimeter and ultra-performance liquid chromatograph(UPLC) were used to determine the color(lightness-shade chromaticity value L~*, red-green chromaticity value a~*, yellow-blue chromati-city value b~*) and chemical components(cynaroside, linarin, luteolin, apigenin, and chlorogenic acid) of 84 CIF germplasms, respectively. Diversity analysis, correlation analysis, regression analysis, and cluster analysis were performed. The results showed that the color and chemical components of CIF were diversified. Chlorogenic acid was in significantly positive correlation with L~* and b~* and significantly negative correlation with a~*. Cynaroside and grey relational grade γ_i of chemical components were in significantly po-sitive correlation with b~* and L~*, respectively, whereas linarin, luteolin, and apigenin had no significant correlation with L~*, a~*, or b~*. The 84 CIF germplasms were clustered into 4 clades. In addition, germplasms in clade Ⅲ had higher γ_i and total color value(E~*_(ab)) than those in other clades, with the best quality and color, and a germplasm with the highest quality, bright yellow color, and highest content of linarin was screened out in this clade. Thus, CIF with bright yellow color had high content of cymaroside and chlorogenic acid and thereby high quality. In summary, the color can be used to quickly predict the quality of CIF. Our results provided data for the evaluation of CIF quality by color and a reference for its procurement and breeding.
Chrysanthemum/chemistry* ; Luteolin/analysis* ; Chlorogenic Acid/analysis* ; Apigenin/analysis* ; Plant Breeding ; Excipients ; Chromatography, High Pressure Liquid/methods*

To define the composition of relevant substances in Breviscapine for Injection, in order to improve the quality control of impurity, and ensure the clinical safety. The analysis and structural identification of relevant substances in different specifications and batches of Breviscapine for Injection powders were carried out by HPLC and UPLC-QTOF-MS. Three primary relevant substances, namely 5,6,7,3',4'-pentahydroxyflavone-7-O-glucuronide(3), 3,5,6,7,4'-pentahydroxyflavone-3-O-glucuronide(4) and scutellarein(10), as well as three minor impurities, namely 6-hydroxyapigenin-6-O-glucosyl-7-O-glucuronide(1), methoxylscutellarin(6) and apigenin-7-O-glucuronide(7) were structurally identified by matching retention time, UV spectra, and mass spectra with authentic compounds and MS fragmentation rules. The main relevant substances(3) and(4) were separated and purified by semi-preparative HPLC, and their structures were further confirmed by NMR data. The study defined relevant substances of Breviscapine for Injection, and provided reference for improving the quality control level of single impurity in breviscapine preparation.
Apigenin/analysis* ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/standards* ; Flavonoids/chemistry* ; Glucuronides/analysis* ; Injections ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Quality Control

Lomatogonium rotatum (L.) Fries, Gentianopsis barbata (Froel) Ma, and Gentianella acuta (Michx.) Hulten, the three kinds of Digeda-species Mongolian medicinal materials belonging to the family Gentianaceae, bad been widely used for the treatment of liver diseases. To analyze comparatively the content of swertiamarin and swertisin among these three kinds of Digeda-species Mongolian medicinal materials. HPLC method was applied for qualitative and quantitative analysis of swertiamarin and swertisin. The Phenomenex C18 (4.6 mm x 250 mm, 5 μm) was used, chromatographic methanol and water as mobile phase, the flow rate was 1.5 mL x min(-1) with UV detected at 237 nm, column oven temperature was 25 degrees C. Results showed that the contents of swertiamarin and swertisin were closely related the different species and producing areas. The content range of swertiamarin in L. rotatum from different habitats was 1.73% - 2.72%, 0.43% - 0.96% for the swertisin content; the content of swertiamarin in G. barbata from Alxa Left Banner was 0.38%, and the content of swertiamarin and swertisin in G. barbata from the others habitats and G. Acuta from different habitats were all detected qualitatively. The contents of swertiamarin and swertisin among these medicinal plants showed a significant difference due to the different species and producing areas. As a consequence, these medicinal plants should not be put together for clinical applications.
Apigenin ; analysis ; Chromatography, High Pressure Liquid ; Gentianaceae ; chemistry ; classification ; Gentianella ; chemistry ; classification ; Iridoid Glucosides ; analysis ; Medicine, Mongolian Traditional ; Mongolia ; Plant Extracts ; analysis ; Pyrones ; analysis

The chemical constituents of 95% ethanol extract of Melastoma dodecandrum were isolated and purified by chromatography on silica gel, Sephadex LH-20, and HPLC, to obtain thirteen compounds eventually. On the basis of their physico-chemical properties and spectroscopic data, these compounds were identified as quercetin (1), quercetin-3-O-β-D-glucopyranoside (2), quercetin-3-O-(6"-O-p-coumaroyl) -β-D-glucopyranoside (3), kaempferol (4), kaempferol-3-O-β-D-glucopyranoside (5), kaempferol-3-O- [2",6"-di-O-(E)-coumaroyl]-β-D-glucopyra-noside (6), luteolin (7), luteolin-7-O-(6"-p-coumaroyl) -β-D-glucopyranoside (8), apigenin (9), apigenin-7-(6"-acetyl-glucopyranoside) (10) , naringenin (11), isovitexin (12), and epicatechin-[8,7-e] -4β-(4-hydroxyphenyl)-3,4-dyhydroxyl-2(3H)-pyranone (13). Eight compounds(3,5,6,8-11 and 13) were obtained from M. dodecandrum for the first time.
Apigenin ; analysis ; Chromatography ; methods ; Chromatography, High Pressure Liquid ; Dextrans ; Flavanones ; analysis ; Flavonoids ; analysis ; chemistry ; Glycosides ; analysis ; chemistry ; Kaempferols ; analysis ; Luteolin ; analysis ; Magnoliopsida ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; analysis ; Silica Gel

OBJECTIVEA high-performance liquid chromatographic (HPLC) method was developed for simultaneous determination of chlorogenic acid, scutellarin, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid in different parts of Erigerontis Herba.

METHODThe four constituents were measured on an Agilent Zorbax SB-C18 column (4.6 mm x 450 mm, 5 microm) with a gradient elution of acetonitrile (A) -0.3% phosphoric acid solution (B) (0-10 min, 12%-15% A, 10-32 min, 15% A, 32-33 min, 15%-20% A, 33-50 min, 20%-22% A) at wavelength of 335 nm and 327 nm, and a flow rate of 1.0 mL x min(-1) and the column temperature was 30 degrees C.

RESULTLinearity of each standard was established in the concentration range of 0.050 1-1.002 microg for chlorogenic acid, 0.165 9-3.318 microg for chlorogenic acid, 0.049 7-0.994 microg for 3,5-dicaffeoylquinic acid, 0.048 7-0.974 p.g for 4,5-dicaffeoylquinic acid respectively, with correlation coefficient r > 0.999 6. Average recoveries (n = 6) of 4 compounds were 98.53% with a RSD of 0.94%, 99.68% with a RSD of 0.49%, 98.78% with a RSD of 1.1%, 99.06% with a RSD of 0.81%, respectively.

CONCLUSIONThe developed method is simple, accurate, and precise, it can be used for the quantitative analysis of Erigeron breviscapus.

Apigenin ; analysis ; chemistry ; Chlorogenic Acid ; analogs & derivatives ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Erigeron ; chemistry ; Glucuronates ; analysis ; chemistry ; Quinic Acid ; analogs & derivatives ; analysis ; chemistry

AIM: To provide a comprehensive procedure to evaluate the contribution of the floral parts to the yield of the major components from the flowers of Trollius chinensis, to underlay the selective breeding, cultivation, development, and utilization of the flowers. METHODS: Five floral parts from eleven batches of the flowers of T. chinensis were examined by HPLC analysis for the content of orientin and vitexin, and by gravimetric analysis for their respective mass fraction. The contribution of each floral part was calculated using mathematical methods based on the results of the content and mass fraction. Variance analysis was carried out by Kruskal-Wallis H test and PCA method. RESULTS: The calculated mean contributions of calyx, corolla, stamens and pistils, stalk, and ovary to the yield of both orientin and vitexin were 76.99% and 71.93%, 9.60% and 8.33%, 9.21% and 8.10%, 2.17% and 6.62%, and 2.03% and 5.02%, respectively. CONCLUSION The floral parts contribute unequally to the yield of orientin and vitexin, and the calyx contributes the highest and makes a significant difference compared with any other part.
Apigenin ; analysis ; Drugs, Chinese Herbal ; analysis ; Flavonoids ; analysis ; Flowers ; chemistry ; Glucosides ; analysis ; Ranunculaceae ; chemistry

This paper aims to establish a new method of calibration and positioning in quantitative analysis of multicomponents by single marker (QAMS), using Shuanghuanglian oral liquid as the research object. Establishing relative correction factors with reference chlorogenic acid to other 11 active components (neochlorogenic acid, cryptochlorogenic acid, cafferic acid, forsythoside A, scutellarin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, baicalin and phillyrin wogonoside) in Shuanghuanglian oral liquid by 3 correction methods (multipoint correction, slope correction and quantitative factor correction). At the same time chromatographic peak was positioned by linear regression method. Only one standard uas used to determine the content of 12 components in Shuanghuanglian oral liquid, in stead of needing too many reference substance in quality control. The results showed that within the linear ranges, no significant differences were found in the quantitative results of 12 active constituents in 3 batches of Shuanghuanglian oral liquid determined by 3 correction methods and external standard method (ESM) or standard curve method (SCM). And this method is simpler and quicker than literature methods. The results were accurate and reliable, and had good reproducibility. While the positioning chromatographic peaks by linear regression method was more accurate than relative retention time in literature. The slope and the quantitative factor correction controlling the quality of Chinese traditional medicine is feasible and accurate.
Apigenin ; analysis ; Calibration ; Chlorogenic Acid ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Flavanones ; analysis ; Flavonoids ; analysis ; Glucosides ; analysis ; Glucuronates ; analysis ; Glycosides ; analysis ; Reproducibility of Results

OBJECTIVETo establish a determination method for contents of chlorogenic acid and vitexin 2"-rhamnoside in Crataegi Fructus extracts by HPLC.

METHODThe chromatographic column was SHISEIDO CAPCELL PAK C18 (4.6 mm x 250 mm, 5 microm); mobile phase was methanol(A) and THF-acetic acid-water (10:2:76, B), with gradient elution, flow speed was 1.0 mL x min(-1); detection wavelength was 330 nm; temperature of column was 25 degrees C.

RESULTChlorogenic acid and vitexin 2"-rhamnoside showed good linear relationships within the ranges of 1.34-164.8 and 1.18-148.7 mg x L(-1), with the recovery rates being 100.4% and 98.8%, and RSD being 1.5% and 1.3% respectively.

CONCLUSIONThe present method is so simply, accurate and highly repeatable that it can be used to effectively determine the contents of chlorogenic acid and vitexin 2"-rhamnoside in Crataegi Fructus extracts.

Apigenin ; analysis ; isolation & purification ; Chlorogenic Acid ; analysis ; isolation & purification ; Chromatography, High Pressure Liquid ; Crataegus ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Medicine, Chinese Traditional ; Plant Extracts ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results

OBJECTIVETo establish an UPLC-MS/MS method for simultaneous determination protocatechuic acid, isoorientin and scutellarin in rat tissues after a single intravenous administration of Compound Hongcao for injection, and investigate the distribution character of the three compounds.

METHODThree compounds were simultaneously determined by UPLC with Waters Acquity UPLC BEH C18 column. The mobile phase, consisting of acetonitrile and 0.1% aqueous formic acid, was programmed as a linear gradient. The flow rate was 0.35 mL x min(-1). The compounds were ionized in the electrospray ionization ion source of the mass spectrometer and detected in the multiple reaction model. The tissue samples were homogenated and the suspension were extracted with methanol for further use.

RESULTThe relationship between the concentration and the peak areas of the three compounds were all linear (r > 0. 99). The precisions, accuracy, extract recoveries and stability of the analytes meet the requirements. The method has been successfully applied to tissue distribution studies of the three compounds. The present study demonstrates that the higher tissues concentration of three components were obtained in kidney, lung and heart after a single intravenous administration of ones.

CONCLUSIONThe present study demonstrates that the three compounds have unequal distribution character. It was showed that the three compounds were mainly distributed in abundant blood-supply tissues such as kidney, lung and heart. It was also found that protocatechuic acid, isoorientin and scutellarin exceretion rapidly and have no long-term accumulation. The method was shown to be effective, convenient, and suitable for simultaneous study the distribution of the three compounds in rat.

Animals ; Apigenin ; analysis ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; pharmacokinetics ; Female ; Glucuronates ; analysis ; Hydroxybenzoates ; analysis ; Injections, Intravenous ; Luteolin ; analysis ; Male ; Rats ; Rats, Wistar ; Tandem Mass Spectrometry ; methods ; Tissue Distribution