iridoids and paeonol

Simultaneous determination method of four marker components, paeoniflorin, loganin, 5-hydroxymethyl-2-furaldehyde and paeonol in traditional herbal medicine, Yukmijihwang tang, was developed to achieve rapidly and systematic quality control by high performance liquid chromatography coupled with diode array detection. To develop this high performance liquid chromatography method, C(18) column (5 microm, 120 A, 4.6 mm x 150 mm) was used with gradient elution of water and methanol as mobile phase. Validation of the chromatography method was evaluated by linearity, recovery, and precision test. Calibration curves of standard components showed excellent linearity with R(2) > 0.9990. Limits of detection and limits of quantification were in the ranges 0.01-0.02 and 0.04-0.07 microg/mL, respectively. The relative standard deviations of data of the intra-day and inter-day experiments were less than 1.15% and 3.76%, respectively. The results of recovery test were found to range from 94.88 to 107.43% with relative standard deviation values 0.07-2.66%. The results of validation suggested that this method was very accurate and stabilized.

Topics: Acetophenones; Benzoates; Bridged-Ring Compounds; Chromatography, High Pressure Liquid; Furaldehyde; Glucosides; Iridoids; Medicine, Korean Traditional; Molecular Structure; Monoterpenes; Plant Preparations; Quality Control; Reference Standards; Reproducibility of Results

To investigate the effects of four original constituents (Loganin, Morroniside, Peoniflorin, Paeonol) of Liu-wei Di-huang pills on the proliferation and differentiation of rat preadipocyte, and optimize the best concentration by the orthogonal test.. The rat preadipocytes were cultured, then above four original constituents according to 4 factors and 3 levels to make L9 (3(4)) orthogonal test, and the control was performed at the same time. The proliferation of preadipocytes was determined by MTT method, and the accumulation of cellular lipid was determined by Oil Red O staining, the differences between factors and variances were compared by the value of absorbance.. The combination of four original constituents of Liu-wei Di-huang pills could stimulate rat preadipocyte proliferation and inhibited its lipid accumulation. Both the effects were highly significant (P < 0.01) with the combination of A3 B1 C3 D2; Compared with the monosomic group, the effects of the A3 B1 C3 D2 was highly significant (P < 0.01).. Compared with the monosomic group, the effects of the combination of the four original constituents of Liu-wei Di-huang pills on the proliferation and differentiation of rat preadipocyte were highly significant. This combination consists of Loganin, morroniside with high doses, Paeonol, Peoniflorin with mid-dose and low-dose, respectively. Regnesent the overall regulatory role of synergy and officiencg.

Topics: Acetophenones; Adipocytes; Animals; Benzoates; Bridged-Ring Compounds; Cell Differentiation; Cell Proliferation; Cells, Cultured; Drug Combinations; Drug Synergism; Drugs, Chinese Herbal; Fats; Glucosides; Glycosides; Iridoids; Male; Monoterpenes; Plants, Medicinal; Rats; Rats, Wistar

On the base of establishing the fingerprint of JinKui ShenQi pills, the ultraviolet spectra-mass spectrometry/mass spectrometry, method was used to identify the fingerprint. Seperation was performed on the Symmetry Shield RP18 (5 microm, 4. 6 mm X 15 mm) analytical column with mobile phase consisting of 1% acetic acid and acetonitrile with gradient elute at the flow rate of 1.0 mL x min(-1), and the ultraviolet detection wavelength was set at 248 nm. Using the above-mentioned chromatographic condition, the fingerprint of different samples was established and the same fingerprint was defined. The fingerprints of different samples were compared with similarity evaluation software published by Pharmacopeia committee codex (2004A). The mass spectrograph with API-ESI ionization source was used, setting the flow rate at 0.5 mL x min(-1) after splitting stream. The pressure of atomization room was 50 Psi, the flow rate of dry gas was 9.0 L x min(-1), the capillary voltage was 4 kV, and the transmission voltage was 70 V. The negative scanner mode was chosen, scan scope was 100-2000, using ion trap to analyze quasimolecular ion peak and the selected fragment ion, and TIC chromatography and second order mass chromatogram were recorded. The major constituents among in JinKui ShenQi pills from different origins were separated well by HPLC. Although there was difference among different origins, they showed nineteen identical characteristic absorption bands. Three fingerprints chemical compositions such as loganin, cinnamal and paeonol were identified based on the retention time and ultraviolet spectra of standard preparation. According to their ultraviolet spectra, molecular weight and fragmentation information, ten peaks in the fingerprint were identified by ultraviolet spectroscopy-mass, spectrometry/massg spectrometry. They are 1,2,3-tri-O-galloyl-glucose, loganin, paeoniflorin, 1,2,3,6-tetro-O-galloyl-glucose, soya-cerebroside, cornuside, and PGG, benzoyl-oxypaeoniflorin. The result showed that the presented fingerprint of JinKui ShenQi pills contained plenty of information quite valuable for the quality evaluation as well as chemical characterization of JinKui ShenQi pills.

Topics: Acetophenones; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Iridoids; Principal Component Analysis; Spectrophotometry, Ultraviolet; Tandem Mass Spectrometry

An HPLC-photodiode array (PDA) detection method was established for the simultaneous determination of 12 components in Xiao-Yao-San-Jia-Wei (XYSJW): geniposide, puerarin, paeoniflorin, ferulic acid, liquiritin, hesperidin, naringin, paeonol, daidzein, glycyrrhizic acid, honokiol, and magnolol. These were separated in less than 70 min using a Waters Symmetry Shield RP 18 column with gradient elution using (A) acetonitrile, (B) water, and (C) acetic acid at a flow rate of 1 ml/min, and with a PDA detector. All calibration curves showed good linear regression (r(2)>0.9992) within the test ranges. The method was validated for specificity, accuracy, precision, and limits of detection. The proposed method enables in a single run the simultaneous identification and determination for quality control of 12 multi-structural components of XYSJW forming the basis of its therapeutic effect.

Topics: Acetophenones; Benzoates; Biphenyl Compounds; Bridged-Ring Compounds; Coumaric Acids; Drugs, Chinese Herbal; Flavanones; Glucosides; Glycyrrhizic Acid; Hesperidin; Iridoids; Isoflavones; Lignans; Medicine, Chinese Traditional; Molecular Structure; Monoterpenes; Quality Control; Reference Standards; Reproducibility of Results; Sensitivity and Specificity

To establish a high performance liquid chromatographic (HPLC) amalgamated to double UV waves method for simultaneous determination of loganin and paeonol in Liuwei Dihuang pills.. A HPLC method was developed. The separation was carried out on a Agilent Zorbax SB C18 column (5 microm, 4.6 mm x 250 mm). The mobile phase consisted of water (A) and acetonitrile (B) with linear linear gradient elution [0-8 min, (B) from 1% to 12%; 8-21 min, B keep 12%; 21-40 min, (B) from 12% to 90%; 40-50 min, B keep 90% for 10 min]. The detection was Photodiode Array with the detection wavelengths were at 236 nm and 274 nm. The column temperature being 30 degrees C and the flow rate was 1.0 mL min(-1). Extracting the chromatergraph from 274 nm and 236 nm, we amalgamated the two chromatographs by matlab programmed.. The calibration curves of loganin and paeonol were linear in the ranges of 0.0362-1.09 microg (r =0. 9998) and 0.0450-1.35 microg (r =0.9998), respectively. The average recoveries of loganin and paeonol were 97.3% (RSD 1.4 %) and 103.0% (RSD 1.9%), respectively. Three different batches of Liuwei Dihuang pills were determined with this method.. This is a more convenient, reasonable and credible quality control method for the traditional Chinese medicine.

Topics: Acetophenones; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Iridoids; Reproducibility of Results; Spectrophotometry, Ultraviolet