Metabolites of Icariin in the Gastrointestine of Rats Following Oral Administration

YinyanghuohasbeenusedasafolkmedicineforthousandsofyearsinChina.ItbelongstothegenusofEpimedium(Berberidaceae).ChinaPharmacopoeia(1995Ed.)collects5speciesofEpimedium,whereflavanoidsareregardedastheprincipalcomponentsresponsibleforthepharmacologicalactivitiesofYinyanghuo,suchasitstonic,antihypertensiveandantiinflammatoryactions.IcariinisoneofthemainflavonoidconstituentsinYinyanghuo"'andisextensivelyusedtocontrolthequalityofthecrudedrug"'.Inpreviouspapers"',wereportedtheisolationandidentificat…     

Determination of Poricoic Acid A in Rat Plasma after Oral and Intravenous Administration by LC–MS–MS and Its Application to a Pharmacokinetic Study

A sensitive and specific liquid chromatography–tandem mass spectrometry (LC–MS–MS) method was developed and validated for the quantification of poricoic acid A (PAA) in rat plasma. The plasma samples were precipitated by protein precipitation with methanol. Glycyrrhetic acid was used as the IS. Chromatography was performed on a Dionex C₁₈ 120 Å (4.6 × 250 mm, 5 μm) column with the mobile phase composed of acetonitrile–water (90:10, v/v) at a flow rate of 0.8 mL min^?1. A tandem mass spectrometer equipped with an ESI source was used as the detector and was operated in the negative ion mode. Quantification was performed using multiple reaction monitoring (MRM) of the transitions m/z 497.4 → 423.3 and m/z 469.2 → 425.1 for PAA and IS, respectively. The calibration curves were linear over the range of 5–5,000 ng mL^?1 (r ² = 0.9966) and the lower limit of quantification (LLOQ) was 5 ng mL^?1. In this range, RSDs were <10% for intra-assay and inter-assay precisions. The accuracy expressed by deviation (DEV) was <6%, and the extraction recoveries of QC samples were >78%. The validated method was successfully used to study the pharmacokinetics of PAA in rats after intravenous administration at a dose of 1.0, 2.5 and 5.0 mg kg^?1 and oral administration at a dose of 25, 50 and 100 mg kg^?1, respectively. The relative bioavailability of PAA in rats following oral administration was achieved.     

LC–MS Determination and Pharmacokinetic Study of Salidroside in Rat Plasma after Oral Administration of Traditional Chinese Medicinal Preparation Rhodiola crenulata Extract

A simple, rapid and sensitive liquid chromatography–mass spectrometry (LC–MS) method was developed for the quantification of salidroside in rat plasma and the study of its pharmacokinetics after oral administration of 15 g kg^?1 Rhodiola crenulata extract to Wistar rats. A 200 μL plasma sample was extracted by acetonitrile and performed on Kromasil C₁₈ column (150 mm × 4.6 mm, 5 μm) with the mobile phase of acetonitrile–water (11:89) within a run time of 8 min. The analyte was monitored with electrospray ionization (ESI) by selected ion monitoring (SIM) mode. The target ions were m/z 299.20 for salidroside and m/z 150.00 for internal standard (IS) paracetamol. A good linear relationship was obtained over the range of 100–20,000 ng mL^?1 and the lower limit of quantification was 100 ng mL^?1. The validated method was successfully applied for the pharmacokinetic study of salidroside in rat. After oral administration of Rhodiola crenulata extract, the main pharmacokinetic parameters T _max, T _1/2, C _max, AUC _0?t and AUC _0?∞ were 0.56 ± 0.21 h, 7.91 ± 4.42 h, 3,386 ± 2,138 ng mL^?1, 16,146 ± 6,558 ng h mL^?1 and 18,599 ± 6,529 ng h mL^?1, respectively.     

Separation and Determination of Methylnaltrexone in Human Plasma Samples After Oral Administration by HPLC Coupled with Electrochemical Detection

Introduction Opioidmedicationsarewidelyusedclinically forrelievingpainandasantidiarrhealsandantitus- sives.Opioidantagonistsconsistofagroupofnat- ural,semisynthetic,orsyntheticcompoundsacting onaseriesofreceptors.Concomitantwiththeabil- itytorelievepain,thesedrugshaveadverseef- fects.Side-effectsofopioidtreatmentincludenau- sea,constipation,urinaryretention,pruritus,fa- tigue,psychomimeticdisturbance,difficultmic- turition,vomitinganddependence.Thesesideef- fectsareoftensevereenoughtolimitthe…     

Simultaneous Determination of Berberine and Palmatine in Rabbit Plasma by LC-MS–MS and Its Application in Pharmacokinetic Study After Oral Administration of Coptidis and Coptidis–Gardeniae Couple Extract

A valid and sensitive LC-MS–MS method is adopted for pharmacokinetics study of berberine and palmatine in rabbit plasma. After mixing with internal standard tetrahydroberberine, plasma samples were pretreated with 1.5 mL acetonitrile. Chromatographic separation was on a C₁₈ column using a mixture of water (containing 10 mmol L^?1 ammonium acetate, pH 3.5) and acetonitrile (50∶50, v/v) as mobile phase. The detection was performed by selected ion monitoring mode via electrospray ionization source operating in the positive ionization mode. The method was linear over the concentration range of 2.0–200.0 ng mL^?1 for berberine and 1.0–100.0 ng mL^?1 for palmatine. The lowest limits of quantitation (LLOQ) were 2.0 ng mL^?1 for berberine and 1.0 ng mL^?1 for palmatine. The intra- and inter-day precision values were less than 14.3% and the deviations were within ±11.0%. The fully validated LC-MS–MS method has been successfully applied to a pharmacokinetic study of berberine, palmatine in rabbit plasma after oral administration of Coptidis and coptidis–gardeniae couple extract. The results indicated that the plasma profiles of the two compounds in rabbit confirmed to one-compartment open model and the combinational utilization with Gardeniae could increase the bioavailability of berberine and palmatine, the two major active components of Coptidis.     

Effect of Temperature-responsive Solution Behavior of PNIPAM-bPPEOMA-b-PNIPAM on Its Inclusion Complexation with α-Cyclodextrin

The effect of temperature-responsive solution behavior of PNIPAM-b-PPEOMA-b-PNIPAM on its inclusion complexation with α-cyclodextrin was studied. The triblock polymer was prepared by reversible addition-fragmentation chain transfer(RAFT) polymerization and formed inclusion complexes(ICs) after selective threading of the PEO segment of the triblock polymer through the cavities of α-CD units. For comparison, PPEOMA homopolymer was prepared and the inclusion complexation with α-CD was also studied. The ICs were prepared with α-CD when the polymer was in different conformations by changing the temperature, and the formed ICs were characterized by XRD, 1H-NMR, TGA and DSC. The solutions of the ICs show temperature-responsive clear/turbid transition or fluidic emulsion/gel transition depending on the concentration of the α-CD added, and the stoichiometry determined by 1H-NMR and TGA indicates that the stoichiometry of EO to α-CD of the resulted ICs increases with increasing of temperature.     

Simultaneous Determination of Pethidine and Atropine in Rabbit Plasma by LC�CMS�CMS and Its Application to a Pharmacokinetic Study after Intramuscular Administration

A sensitive and selective liquid chromatography?Ctandem mass spectrometry method for the determination of pethidine and atropine in rabbit plasma was developed and validated. The analytes and internal standard (IS) are extracted from plasma by liquid?Cliquid extraction using ethyl acetate, and separated on a Zorbax SB-Aq column (2.1 × 150 mm, 3.5 ??m) using acetonitrile?C0.1% formic acid as mobile phase with gradient elution. Electrospray ionization source was applied and operated in positive ion mode, and multiple reaction monitoring mode was used for quantification using target fragment ions m/z 247.8 ?? 219.7 for pethidine, m/z 289.9 ?? 123.8 for atropine and m/z 295.0 ?? 266.8 for IS, respectively. The assay is linear over the range of 5?C1,000 ng mL^?1 for pethidine and atropine, with a lower limit of quantification of 3 ng mL^?1 for pethidine and 5 ng mL^?1 for atropine. Intra-day and inter-day precision are less than 11% and the accuracy are in the range of 90.4?C106.3%. Furthermore, the newly developed method is successfully used for the determination of pethidine and atropine in rabbit plasma for pharmacokinetic study.     

Development and Validation of a LC–ESI–MS Assay for Determination of Icariin in Rat Plasma after Administration of Herba Epimedii

This paper reports a sensitive, specific, and stable chromatographic procedure with selective detection (electrospray mass spectrometry in selected-ion-monitoring mode) combined with simple and efficient sample preparation for determination of icariin in rat plasma after administration of Herba Epimedii. Separation of the analyte, possible endogenous compounds, and constituents of Herba Epimedii were accomplished on a 250 mm × 2.0 mm i.d. C₁₈ column by use of a rapid gradient. Ionization of icariin and clarithromycin (internal standard) was achieved by use of the electrospray interface in positive-ion mode. Conditions such as ionization mode, type of organic modifier, eluent additives, and Q-array potential were optimized to achieve good sensitivity and specificity of icariin detection. Response was a linear function of concentration over the range 0.2–20 ng mL⁻¹. The method is accurate and precise; within-batch and between-batch precision (CV) are <15%, and accuracy (RE) is better than ±15%. The method can be used for analysis of icariin in plasma after administration of Herba Epimedii or of traditional Chinese medicinal preparations containing Herba Epimedii.     

Development of an LC Method for Simultaneous Analysis of Cinnamic Acid and Paeonol in Rat Plasma, and Its Application to a Pharmacokinetic Study after Intragastric Administration of Guizhi–Fuling Capsule

A specific and accurate high-performance liquid chromatographic method for analysis of cinnamic acid (CA) and paeonol (PN) in rat plasma has been developed and validated. Plasma samples were pretreated by protein precipitation with methanol, and the supernatant was injected for reversed-phase separation on a C₁₈ column with acetonitrile–0.1% phosphoric acid 24:76 (v/v) as mobile phase at a flow-rate of 1.0 mL min^?1. Phenylbutyric acid was used as the internal standard. Good linear relationships were obtained between response and concentration in the range 0.130–52.0 μg mL^?1 (r = 0.9980) and 0.1785–71.4 μg mL^?1 (r = 0.9950) for CA and PN, respectively. Intra-day and inter-day assay precision (RSD, n = 6) at three concentrations was not above 15.1% for either CA and PN, and accuracy was from 94.3 to 104.7% and from 103.3 to 113.1% for CA and PN, respectively. Mean recovery of CA and PN from plasma samples was 87.5 and 86.8%, respectively, and recovery of the internal standard at a concentration of 1.00 mg mL^?1 was 88.5%. Results from a stability study suggested CA and PN were stable under the experimental conditions used. Finally, the validated method was successfully applied to a pharmacokinetic study of CA and PN in rats after intragastric administration of Guizhi–Fuling capsule. The results obtained would be very useful for evaluation of the clinical efficacy of GFC.     

Thermodynamics of Binding of α:-Cyclodextrin to Straight-Chain Alkyl Derivatives in Aqueous Solution

Apparent standard Gibbs energy, enthalpy, entropy, and heatcapacity data of the interactions of -cyclodextrin (CD) to some n-carboxylatesH(CH₂)_nCOO^- (n = 4–6), are determined by isothermal titration microcalorimetryat different temperatures in phosphate buffer, pH 9.0, assuming a 1 : 1 model indilute solution. Modelling of contributions of the thermodynamic properties of the solutionindicates that CD undergoes conformational change upon binding to homologousseries of n-carboxylates, n-alcohols, ,-alkane dicarboxylates and ,-alkane diols.     

LC-MS Determination and Pharmacokinetic Study of Luteolin-7-O-β-d-glucoside in Rat Plasma after Administration of the Traditional Chinese Medicinal Preparation Kudiezi Injection

A rapid and sensitive LC-MS method has been developed for the determination of luteolin-7-O-β-d-glucoside in rat plasma after solvent extraction. Separation was on an Elite Hypersil ODS2 column (250 mm × 4.6 mm i.d., 5 μm) with a mobile phase of acetonitrile-0.3% acetic acid (26:74, v/v). The samples were analyzed by using positive electrospray ionization MS in selected ion monitoring mode. The selected ions for luteolin-7-O-β-d-glucoside and the internal standard, isoquercitrin, were m/z 448.95 and m/z 464.95. Good linearity was observed over the range of 20–2,000 ng mL^?1 with a lower limit of quantification of 20 ng mL^?1. No interference peaks or matrix effects were observed. The validated method was applied to the pharmacokinetic study of luteolin-7-O-β-d-glucoside in rat plasma after intravenous administration of Kudiezi Injection.     

Development and Validation of Highly Sensitive LC–ESI-MS/MS Method for Bortezomib and Its Applications for Plasma Levels and Drug Content of Branded and Generic Formulations in India

In the present study, a highly sensitive and reproducible bio-analytical method was developed using LC–ESI-MS/MS to assess the lower plasma levels of bortezomib in multiple myeloma patients. The gradient elution was optimized using reverse-phase C18 column with mobile phases consisting of water and acetonitrile in 0.1% formic acid. Multiple reaction monitoring mode was used for quantification using precursor-to-product ion transition for bortezomib and sulfadiamethoxine was used as internal standard. This method was validated with a linearity range of 0.195–25 ng mL^?1. Intra-day and inter-day accuracy was 99.17–101.89% and 95.01–102.92% with precision of?<?9.87% and?<?8.77%, respectively. Bortezomib was stable in plasma samples stored at ? 80 °C for up to 10 months. The lower limit of quantification was found to be 0.195 ng mL^?1. This method was also found to be capable of quantifying bortezomib trough levels (ranging 0.19–0.7 ng mL^?1) in plasma of multiple myeloma patients post-cycle 1–6. Bortezomib content in the commonly prescribed generic formulations was also studied. The concentration in all formulations was within the 90–110% of the innovator, as prescribed by the USFDA, ruling out their role blood level variation. The study supports the use of this method for trough level estimation and therapeutic drug monitoring of bortezomib in multiple myeloma patients.

     

LC–ESI–MS Determination of Imperatorin in Rat Plasma After Oral Administration and Total Furocoumarins of Radix Angelica dahuricae and its Application to a Pharmacokinetic Study

A simple, rapid, sensitive and reliable liquid chromatography–electrospray ionization mass spectrometry method for the quantification of imperatorin in rat plasma after oral administration and total furocoumarins of Radix Angelica dahuricae has been established. The plasma samples were deproteinized by adding internal standard (IS) osthole solution, which was prepared by acetonitrile. The analysis was performed on a Shim-pack C₁₈ column (150 × 2.0 mm i.d., 5 μm) using acetonitrile and 0.5% formic acid solution (70:30, v/v) as a mobile phase. The detection was performed on a quadrupole mass spectrometer detector with an ESI interface operated in the selected ion monitoring mode. The linear quantification range of the method was 2–4000 ng mL^?1 in rat plasma with a correlation coefficient greater than 0.99, the limit of detection (LOD) was 0.5 ng mL^?1 and the lower limit of quantification (LLOQ) 2 ng mL^?1. The intra- and inter-day relative standard deviations (RSD) were less than 2.5 and 3.5%, respectively. The recoveries were above 90%. The validated method was successfully applied to a pharmacokinetic study of imperatorin in rats after oral administration and total furocoumarins of Radix Angelica dahuricae.     

Blood Levels of Hexavalent Chromium in Rats. “In Vitro” and “In Vivo” Experiments

Abstract

For the Cr(VI) selective separation from biological materials we have developed a highly rapid extraction-separation method with liquid anion exchanger as Amberlite LA-1 or LA-2. The analytical determination of Cr(VI) in organic phase was carried out using electrothermal atomic absorption spectroscopy (ETA-AAS).

After i.v. administration of 0.5 and 2.5mg/kg b.w. of K₂Cr₂O₇ in male Wistar rats the biological samples, collected at different times, were immediately analyzed. Cr(VI) was not detected in whole blood one minute after administration of the lower dose. In blood of rats receiving higher dose an incomplete reduction of Cr(VI) was observed.

Such data demonstrate a highly rapid but limited metabolic capacity of hematic compartment to reduce Cr(VI) to trivalent status.

These results obtained with a new and specific analytical method, confirmed a trigger role of red cells in Cr(VI) metabolism.

“In vitro” incubation of K₂Cr₂O₇ (4 μM) with rat erythrocytes or plasma at 37°C showed a rapid reduction of Cr(VI) in red cells while plasma samples demonstrated a limited reductive power.     

Identification of Acepromazine and Its Metabolites in Horse Plasma and Urine by LC–MS/MS and Accurate Mass Measurement

Acepromazine maleate (Sedalin^®) was administered orally to six thoroughbred horses at a dose of 0.15 mg kg⁻¹. Urine and blood samples were collected up to 412 h post-administration. Plasma and urine were hydrolysed; plasma samples were then processed using liquid–liquid extraction and urine samples using solid-phase extraction. A sensitive tandem mass spectrometric method was developed in this study, achieving a lower limit of quantification for acepromazine of 10 pg mL⁻¹ in plasma and 100 pg mL⁻¹ in urine. Acepromazine, hydroxyethylpromazine, hydroxyacepromazine, hydroxyethylpromazine sulphoxide, hydroxyethylhydroxypromazine, dihydroxyacepromazine and dihydroxyhydroxyethylpromazine were detected in the post-administration samples. The parent drug and its metabolites were identified using a combination of UPLC–MS/MS and accurate mass measurement. Separation of the structural isomers hydroxyethylpromazine sulphoxide and hydroxyethylhydroxypromazine was another significant outcome of this work and demonstrated the advantages to be gained from investing in chromatographic method development.

     

Identification of Acepromazine and Its Metabolites in Horse Plasma and Urine by LC–MS/MS and Accurate Mass Measurement

Acepromazine maleate (Sedalin^?) was administered orally to six thoroughbred horses at a dose of 0.15?mg?kg^?1. Urine and blood samples were collected up to 412?h post-administration. Plasma and urine were hydrolysed; plasma samples were then processed using liquid–liquid extraction and urine samples using solid-phase extraction. A sensitive tandem mass spectrometric method was developed in this study, achieving a lower limit of quantification for acepromazine of 10?pg?mL^?1 in plasma and 100?pg?mL^?1 in urine. Acepromazine, hydroxyethylpromazine, hydroxyacepromazine, hydroxyethylpromazine sulphoxide, hydroxyethylhydroxypromazine, dihydroxyacepromazine and dihydroxyhydroxyethylpromazine were detected in the post-administration samples. The parent drug and its metabolites were identified using a combination of UPLC–MS/MS and accurate mass measurement. Separation of the structural isomers hydroxyethylpromazine sulphoxide and hydroxyethylhydroxypromazine was another significant outcome of this work and demonstrated the advantages to be gained from investing in chromatographic method development.