ABSTRACT
To study the metabolic products of main compounds of Chuankezhi injection in rat, 12 Sprague Dawley rats were classed into 2 groups, a blank control group and an intermuscular administration group, respectively. Rat feces and urine samples were collected from 0-24 h and 24-48 h after administration. All the samples were ultrasonically treated with methanol and then analyzed using LC-LTQ Orbitrap MSn. By comparison with the total ion chromatogram of samples from the blank control group, the metabolites in the samples of drug-treated group were screened. These metabolites were further analyzed by multistage product ion scanning and comparison of retention time with reference substances. As a result, a total of 12 flavonoid metabolites were tentatively identified from the rat feces and no metabolite was discovered in the rat urine. Epimedin C and icariin were detected in the rat blood samples after 30 min of administration, but their metabolites and other original flavones were not detected. Furthermore, no original flavones and their metabolites were detected in rat blood samples after 2 and 4 h of administration. The potential metabolism paths were further characterized and the principal in vivo transformation of flavones from Chuankezhi injection were deglycosylation, dehydration, methylation, oxidation and isomerization in rats.
ABSTRACT
A quantitative method for epimedin A, B, C and icariin in rat plasma was established using LC-MS/MS after intermuscular administration of Chuankezhi injection to rat. Chromatographic separation was performed on an Agilent Eclipse XDB-C18 column (150 mm×2.1 mm, 5.0 μm) at 40℃. Mobile phase consisted of acetonitrile -0.1% formic acid in water (35:65), and the flow rate was 0.22 mL·min-1. The LC effluent was detected and analyzed using an ESI-triple quadrupole tandem mass spectrometer under the multiple reaction monitoring (MRM) in the negative ion mode. The plasma samples were treated with solid phase extraction prior to LC-MS/MS analysis. As a result, all of the four analytes displayed a good linearity over the concentration of 1-1000 ng·mL-1. The RSDs of intra-day and inter-day assays were less than 5.99% and 10.16%, respectively. The relative recovery of each analyte was between 88.1%-101.1% with RSD<7.9% and the absolute recovery was between 72.0%-86.6% (RSD<6.3%). In conclusion, the established method shows good specificity, sensitivity and efficiency for quantifying the four flavonoid glycosides contained in rat plasma.
ABSTRACT
To study pharmacokinetic characteristics of epimedin A, B, C and icariin after intermuscular administration of Chuankezhi injection to rat. The established RRLC-MS/MS method was applied for simultaneous determination of four analytes in rat plasma and calculating their pharmacokinetic parameters. As a result, each analyte showed a good linear relationship in the concentration range of 1-1 000 μg•L⁻¹.The intra-day precise was 96.9%-107.5% with RSD<5.99%, inter-day precise was 92.3%-105.0% with RSD<10.16%. The relative recovery of four analytes was 88.1%-101.1% with RSD<7.9% and their absolute recovery was 72.0%-86.6% with RSD<6.3%. After intermuscular administration of Chuankezhi injection, the plasma concentration of four flavonoid glycosides rapidly arose to peaks at about 10 min, and then quickly declined in rat. Tmax of epimedin A, B, C and icariin was 0.21, 0.19, 0.16 and 0.49 h, respectively, and their mean elimination half-life(t1/2z) was 0.60, 0.62, 0.47 and 0.49 h. The established method was validated to be sensitive, rapid and specific for determination of the four analytes. Serum concentration of 4 species of epimedium flavonoids in Chuankezhi injection was low, and their absorption and elimination seem quickly, displaying similar pharmacokinetic characteristics in this study.
ABSTRACT
In this study, an analytical method was developed and used to quantify simultaneously protocatechuic acid, neochlorogenic acid, cryptochlorogenic acid and 1, 3-dicaffeoylquinic acid--four bioactive compounds contained in Fructus Xanthii using UPLC. The contents of four phenolic components of 28 batches of samples collected from different product areas and markets were determined and compared by means of this established method. The mobile phase was composed of methanol and water containing 0.1% phosphoric acid. Chromatography was monitored at dual-wavelengths--220 and 327 nm. Flow rate was 0.4 mL x min(-1) and column temperature was 35 degrees C. The correlation coefficient between concentration and chromatographic peak area of protocatechuic acid, neochlorogenic acid, cryptochlorogenic acid and 1, 3-dicaffeoylquinic acid was over 0.9999 in the range of 0.3570-35.70, 2.500-250.0, 1.060-106.1, 1.010-101.0 microg x mL(-1), respectively. The average recoveries of the four compounds were 97.68%, 99.55%, 97.92% and 100.4%, respectively. In conclusion, the established method can rapidly attain an accurate and reproducible result used to control the quality of Fructus Xanthii.
Subject(s)
Chlorogenic Acid , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Fruit , Chemistry , Hydroxybenzoates , Plants, Medicinal , Chemistry , Quality Control , Quinic Acid , Reproducibility of Results , Xanthium , ChemistryABSTRACT
To establish a sensitive and specific HPLC method for quality control of Radix Paeoniae Alba, HPLC method was applied for quality assessment of Radix Paeoniae Alba. HPLC analysis was performed on a Symmetry C18 column (250 mm x 4. 6 mm ID, 5 microm, Waters, USA). The mobile phase consisted of acetonitrile (solvent A) and water containing 0.1% (v/v) phosphoric acid (solvent B) at a constant flow rate of 0.8 mL x min(-1). An increasing linear gradient (v/v) of solvent A was used (t/min, % A): (0,10), (5,10), (25,15), (45, 22), (46, 65), (50, 80) and (60, 80). The column temperature was set at 25 degrees C. The chromatograms were monitored at 230 nm and the on-line UV spectra were recorded in the range of 190 - 400 nm. The HPLC chromatographic fingerprinting of Radix Paeoniae Alba, showing 11 characteristic peaks, was established from 28 lots of Radix Paeoniae Alba. The areas of main chromatographic peaks were found to complied with the following rule: paeoniflorin > 1, 2, 3, 4, 6-penta-O-galloyl-glucos > albiflorin > methyl gallate > other compounds. The chromatographic fingerprinting of Radix Paeoniae Alba with high specificity can be used to control its quality and assure lot-to-lot consistency.
Subject(s)
Benzoates , Bridged-Ring Compounds , China , Chromatography, High Pressure Liquid , Methods , Ecosystem , Glucosides , Mass Spectrometry , Methods , Monoterpenes , Paeonia , Chemistry , Plant Roots , Chemistry , Plants, Medicinal , Chemistry , Reproducibility of ResultsABSTRACT
<p><b>AIM</b>To analyze the chemical constituents of cortex moutan by liquid chromatography coupled with electrospray ionization mass spectrometry.</p><p><b>METHODS</b>An on-line optimized HPLC-DAD/MS2 technique was employed.</p><p><b>RESULTS</b>In the negative ion detection mode, 38 components such as monoterpene glucosides, galloylglucoses and acetophenones were isolated. Among them, over thirty compounds were identified, including paeonol, paeonilflorin, oxypaeoniflorin, benzoylpaeoniflorin, benzoyloxypaeoniflorin, galloylpaeoniflorin, galloyloxypaeoniflorin, mundanpioside A, C, D, E, H, etc. by the high performance liquid chromatography with diode-array detection in parallel with electrospray ionization and quadrupole-time of flight tandem mass spectrometry (HPLC-DAD/ESI-MS2).</p><p><b>CONCLUSION</b>This method can be used to rapidly determine the constituents of cortex moutan.</p>
Subject(s)
Acetophenones , Chemistry , Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Chemistry , Glucosides , Chemistry , Molecular Structure , Monoterpenes , Chemistry , Paeonia , Chemistry , Plant Extracts , Chemistry , Plants, Medicinal , Chemistry , Reproducibility of Results , Spectrometry, Mass, Electrospray Ionization , Methods , Tandem Mass Spectrometry , MethodsABSTRACT
<p><b>OBJECTIVE</b>To establish a HPLC fingerprint for quality evaluation of Cortex Moutan.</p><p><b>METHOD</b>The HPLC chromatographic fingerprinting of 30 lots of Cortex Moutan was established and major peaks were identified by LC-MS and MS-MS.</p><p><b>RESULT</b>The HPLC fingerprint of Cortex Moutan was established, showing 15 characteristic peaks. The areas of these peaks were found to complied with the following rule: paeonol > 1, 2, 3, 4, 6-penta-O-galloyl-glucos > methyl gallate > galloylpaeoniflorin > gallic acid > oxypaeoniflorin > other compounds.</p><p><b>CONCLUSION</b>The chromatographic fingerprinting of Cortex Moutan with high specificity can be used to control its quality and monitor lot to lot consistency.</p>
Subject(s)
Acetophenones , Bridged Bicyclo Compounds, Heterocyclic , Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Chemistry , Classification , Gallic Acid , Glucosides , Hydrolyzable Tannins , Monoterpenes , Paeonia , Chemistry , Classification , Plant Bark , Chemistry , Plants, Medicinal , Chemistry , Quality ControlABSTRACT
<p><b>AIM</b>To study the metabolic pathways of ginsenoside Rd in rats.</p><p><b>METHODS</b>Urine samples were collected before and after 24 h of single oral administration of 150 mg and intravenous administration of 60 mg of ginsenoside Rd to six rats, separately. The samples were purified by SPE column and then were analyzed by liquid chromatography-ESI-mass spectrometry for putative metabolites.</p><p><b>RESULTS</b>Parent drug and its seven metabolites were identified in rat urine based on comparing total ion chromatograms of the blank with the metalolic urine as well as mass spectra. Its main metabolic pathways and possible structures are elucidated.</p><p><b>CONCLUSION</b>Oxidation, combination and deglucosylation were found to be the major metabolic pathway of ginsenoside Rd in rats.</p>