Xiangyang Wang

The effect of Compound Danshen Dripping Pills, a Chinese herb medicine, on the pharmacokinetics and pharmacodynamics of warfarin in rats

AIM OF THE STUDY Significant pharmacokinetic/pharmacodynamic (PK/PD) interactions between various herbal products and warfarin have recently been reported. The present study was conducted to determine whether Compound Danshen Dripping Pills (CDDP), a Chinese herb medicine used for the treatment of cardiovascular diseases, interacts with warfarin when administered concomitantly. MATERIALS AND METHODS Each day for 7 days two groups of rats were treated orally with CDDP (50mg/kg and 250 mg/kg, twice daily), and the control group received similar treatment with appropriate volumes of water only. Sixty minutes after the final daily administration of CDDP or water, an aqueous solution of warfarin (0.2mg/mL) was given to each rat at a dose of 1.0mg/kg, and blood samples were collected at 0, 0.5, 1, 2, 4, 8, 12, 24, 36, and 48 h after warfarin-treatment. The concentration of warfarin in blood plasma was determined by high performance liquid chromatography (HPLC). Prothrombin time (PT) in blood plasma was measured using thromboplastin reagent. RESULTS Excellent linearity was found between 0.05 and 10 μg/mL with a lower limit of quantitation (LLOQ) of 0.05 ng/mL (r>0.999); moreover, all the validation data including accuracy and precision (intra- and inter-day), were within the required limits. No significant differences were found in PT(max) and AUC(PT0-∞) between the two CDDP-treated groups and the control. Besides, there was little alteration in any of the pharmacokinetic parameters of warfarin between the two CDDP-treated groups and the control. CONCLUSION The concomitant application of CDDP and warfarin did not give rise to significant effect on the pharmacodynamics of warfarin, and practically no effect on its pharmacokinetics. It was speculated that the PK/PD interactions between CDDP and warfarin was likely to be negligible as long as the patients took CDDP at a normal dose.

Simultaneous determination of five phenolic components and paeoniflorin in rat plasma by liquid chromatography-tandem mass spectrometry and pharmacokinetic study after oral administration of Cerebralcare granule(®).

A simple, sensitive and selective high-performance liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) method was developed for simultaneous determination and pharmacokinetic study of six active components, protocatechuic acid, chlorogenic acid, caffeic acid, ferulic acid rosmarinic acid and paeoniflorin in rat plasma after oral administration of Cerebralcare granule(®) for the first time. The method involves a simple liquid-liquid extraction with ethyl acetate. The separation was performed on a Luna C18 column (2.0×100mm i.d., 3.0μm, particle, Phenomenex, USA) with gradient elution using a mobile phase composed of acetonitrile and water (containing 0.1% formic acid) at a flow rate of 0.2ml/min. Electrospray ionization (ESI) in negative ion mode and selective reaction monitoring (SRM) was used for the quantification of six active components and internal standard (IS, Chloroamphenicol). The method was linear for all analytes over investigated range with all correlation coefficients greater than 0.9914. The lower limits of quantification (LLOQ) were 1.0ng/ml for protocatechuic acid, 1.0ng/ml for chlorogenic acid, 1.0ng/ml for caffeic acid, 5.0ng/ml for ferulic acid, 1.5ng/ml for rosmarinic acid and 6.0ng/ml for paeoniflorin, respectively. The intra- and inter-day precisions (R.S.D.%) were less than 6.60% and 11.68%, and accuracy (RE %) between -3.26% and 1.13% (n=6). The developed method was applied for the first time to the pharmacokinetic study of protocatechuic acid, chlorogenic acid, caffeic acid, ferulic acid, rosmarinic acid and paeoniflorin in rat plasma after oral administration of Cerebralcare granule(®).

Determination of ginsenoside Rc in rat plasma by LC-MS/MS and its application to a pharmacokinetic study.

Ginsenoside Rc (GRc) is a potential pharmacologically active ingredient isolated from ginseng (Panax ginseng C.A. Meyer, Araliaceae). A simple, rapid and sensitive method for determination of GRc in rat plasma was developed based on liquid chromatography-tandem mass spectrometry (LC-MS/MS). The analyte and internal standard (IS), ginsenoside Rb(1) (GRb(1)), were extracted from plasma with n-butanol and chromatographied on a C(18) column eluted with a mobile phase of methanol and water containing 0.1% formic acid. The detection was performed by positive ion electrospray ionization in selective reaction monitoring mode (SRM), monitoring the transitions m/z 1101.6→789.3 and m/z 1131.7→364.7 for GRc and IS, respectively. The assay was linear over the concentration range of 5-5000 ng/mL with a limit of quantitation (LOQ) of 5 ng/mL. The accuracy was between 86.7% and 114.9%, and the precision was less than 9.7%. This method was successfully applied to investigate the pharmacokinetic study of GRc in rats after intravenous (2mg/kg) and oral (20mg/kg) administration, and the result showed that the ginsenoside was poorly absorbed with an absolute bioavailability being approximately 0.17%.

Simultaneous determination of caffeic acid and its major pharmacologically active metabolites in rat plasma by LC-MS/MS and its application in pharmacokinetic study.

A simple, sensitive and selective high-performance liquid chromatography electrospray ionization tandem mass spectrometry (LC-MS/MS) method was developed for simultaneous determination and pharmacokinetic study of caffeic acid (CA) and its active metabolites. The separation with isocratic elution used a mobile phase composed of methanol and water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min. The detection of target compounds was done in selected reaction monitoring (SRM) mode. The SRM detection was operated in the negative electrospray ionization mode using the transitions m/z 179 ([M - H](-) ) → 135 for CA, m/z 193 ([M - H](-) ) → 134.8 for ferulic acid and isoferulic acid and m/z 153 ([M - H](-) ) → 108 for protocatechuic acid. The method was linear for all analytes over the investigated range with all correlation coefficients 0.9931. The lower limits of quantification were 5.0 ng/mL for analytes. The intra- and inter-day precisions (relative standard deviation) were <5.86 and <6.52%, and accuracy (relative error) was between -5.95 and 0.35% (n = 6). The developed method was applied to study the pharmacokinetics of CA and its major active metabolites in rat plasma after oral and intravenous administration of CA.

Simultaneous determination of creatine phosphate, creatine and 12 nucleotides in rat heart by LC–MS/MS

A simple, rapid and sensitive LC-MS/MS method was developed and validated for simultaneous determination of creatine phosphate (CP), creatine (Cr) and 12 nucleotides in rat heart. The analytes, ATP, ADP, AMP, GTP, GDP, GMP, CTP, CDP, CMP, UTP, UDP, UMP, CP, Cr, were extracted from heart tissue with pre-cooled (0°C) methanol/water (1:1, v/v) and separated on a Hypersil Gold AQ C18 column (150mm×4.6mm, 3μm) using an isocratic elution with a mobile phase consisting of 2mmol/L ammonium acetate in water (pH 10.0, adjusted with ammonia). The detection was performed by negative ion electrospray ionization in selective reaction monitoring mode (SRM). In the assay, all the analytes showed good linearity over the investigated concentration range (r>0.99). The accuracy was between 80.7% and 120.6% and the precision expressed in RSD was less than 15.6%. This method was successfully applied to measure the concentrations of the 12 nucleotides, creatine phosphate and creatine in rat heart for the first time.

UFLC–MS/MS determination and pharmacokinetic studies of six Saikosaponins in rat plasma after oral administration of Bupleurum Dropping Pills

A rapid and sensitive ultra fast liquid chromatography tandem mass spectrometry method (UFLC-MS/MS) was developed and validated for the simultaneous determination of six Saikosaponins (SSs) (SSa, SSb1, SSb2, SSd, SSc, SSf) of Bupleurum Dropping Pills (BDP) in rat plasma using chloramphenicol as the internal standard (IS). The SSs were separated using an ACQUITY UPLC(®) BEH C18 column (50 mm × 2.1mm, 1.7 μm) and detection of these compounds were done by using a Qtrap 5500 mass spectrometer coupled with negative electrospray ionization (ESI) source under the multiple reaction monitoring (MRM) mode. According to regulatory guidelines, the established method was fully validated and results were showed within acceptable limits. The lower limit of quantifications (LLOQs) of all analytes were 0.2 ng/mL. The validated method was successfully applied into a pharmacokinetic study of orally administered BDP in rats.

Simultaneous determination of ascaridole, p-cymene and α-terpinene in rat plasma after oral administration of Chenopodium ambrosioides L. by GC-MS

A sensitive and reliable GC-MS method was developed and validated for the simultaneous determination of ascaridole, p-cymene and α-terpinene in rat plasma using naphthalene as internal standard. The plasma samples were extracted with ethyl acetate. Chromatographic separation was carried out on a HP-5MS capillary analytical column (30 m × 0.25 mm, 0.25 µm) and detection was performed on a quadrupole mass spectrometer detector operated under selected ion monitoring mode. The method showed excellent linearity over the investigated concentration range (r > 0.99) with the limit of quantitation down to 50, 10 and 5 ng/mL for ascaridole, p-cymene and α-terpinene, respectively. The intra-day and inter-day precisions (RSD) were <11.3%, and the accuracy was between 90.7 and 113.8%. The method was successfully applied to investigate the pharmacokinetics of Chenopodium ambrosioides L. following oral administration to rats.

Simultaneous determination and pharmacokinetics of danshensu, protocatechuic aldehyde, 4-hydroxy-3-methyloxyphenyl lactic acid and protocatechuic acid in human plasma by LC–MS/MS after oral administration of Compound Danshen Dripping Pills

Graphical abstract The pharmacokinetic study of DSS, PCA and their related metabolites HLMA and PAA in Chinese healthy volunteers after oral administration of CDDP Figure. No Caption available. HighlightsDevelopment of an LC‐MS/MS method for simultaneous determination of DSS,PCA,HMLA and PAA.Application of the LC‐MS/MS method for determination of the four analytes in human plasma.The first pharmacokinetics study of the four analytes in human after oral administration of Compound Danshen Dripping Pills(CDDP). Abstract Compound Danshen Dripping Pills (CDDP), a herbal patent medicine, is widely used in China for the prevention and treatment of cardiovascular diseases. A simple, sensitive and reliable method for simultaneous determination of danshensu (DSS), protocatechuic aldehyde (PCA), and their related metabolites, 4‐hydroxy‐3‐methyloxyphenyl lactic acid (HMLA) and protocatechuic acid (PAA) in human plasma was developed and validated based on liquid chromatography tandem mass spectrometry (LC–MS/MS). The analytes and internal standard (IS), vanillic acid (VAA), were extracted from plasma with ethyl acetate and separated on a C18 column by using the mobile phase consisted of methanol‐0.1% formic acid via gradient elution. The electrospray ionization (ESI) source was applied and operated under the multiple reaction monitoring (MRM) mode. The linear calibration curves were obtained at the concentration ranges of 0.46–1000 ng/mL for DSS and PAA, and 1.38–1000 ng/mL for PCA and HMLA, respectively. The inter‐ and intra‐day precisions (RSD%) were less than 13.5%, and the accuracy (±RE%) was within 13.4%. The described method was successfully applied for the clinical pharmacokinetics of CDDP in Chinese healthy volunteers.

Simultaneous Determination and Pharmacokinetic Study of Protocatechuic Aldehyde and Its Major Active Metabolite Protocatechuic Acid in Rat Plasma by Liquid Chromatography-Tandem Mass Spectrometry.

A very simple and selective high-performance liquid chromatography electrospray ionization tandem mass spectrometry (LC-MS-MS) method was developed for simultaneous determination and pharmacokinetic study of protocatechuic aldehyde (PAL) and its active metabolite protocatechuic acid (PCA). The method involves a simple liquid-liquid extraction with ethyl acetate. The separation was performed on a Hypersil GOLD C18 column (2.1 × 150 mm, 3.0 µm; particle, Thermo, USA) with isocratic elution using a mobile phase consisted of methanol and water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min. The detection of target compounds was done by using low-energy collision dissociation tandem mass spectrometry (CID-MS-MS) using the selective reaction monitoring scan mode. The method was linear for all analytes over the investigated range for all correlation coefficients greater than 0.9950. The lower limits of quantification were 2.0 ng/mL for PAL and PCA. The intra- and interday precisions (relative standard deviation, RSD %) were <6.84 and 5.54%, and the accuracy (relative error, RE %) was between -2.85 and 0.74% (n = 6). The developed method was applied to study the pharmacokinetics of PAL and its major active metabolite PCA in rat plasma after oral and intravenous administration of PAL.

Quantitative determination of periplocymarin in rat plasma and tissue by LC‐MS/MS: Application to pharmacokinetic and tissue distribution study

A simple, rapid and sensitive liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for the determination of periplocymarin in biological samples was developed and successfully applied to the pharmacokinetic and tissue distribution study of periplocymarin after oral administration of periplocin. Biological samples were processed with ethyl acetate by liquid-liquid extraction, and diazepam was used as the internal standard. Periplocymarin was analyzed on a C18 column with isocratic eluted mobile phase composed of methanol and water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min (73:27, v/v). Detection was performed on a triple-quadrupole tandem mass spectrometer using positive-ion mode electrospray ionization in the selected reaction monitoring mode. The MS/MS ion transitions monitored were m/z 535.3→355.1 and 285.1→193.0 for periplocymarin and diazepam, respectively. Good linearity was observed over the concentration ranges. The lower limit of quantification was 0.5 ng/mL in plasma and tested tissues. The intra-and inter-day precisions (relative standard deviation) were <10.2 and 10.5%, respectively, and accuracies (relative error) were between -6.8 and 8.9%. Recoveries in plasma and tissue were >90%. The validated method was successfully applied to the pharmacokinetic and tissue distribution studies of periplocymarin in rats. Copyright