Determination of Jasminoidin in Rabbit Plasma for the Pharmacokinetic Investigation after Single Dose Oral Administration of Gardenia jasminoides Ellis and Gardenia jasminoides Ellis Coupling Coptis chinensis Franch Extracts

A simple, sensitive and rapid method for the analysis of jasminoidin in rabbit plasma by liquid chromatography coupled to tandem mass spectrometry was developed. Detection was by positive ion electrospray ionization in multiple reactions monitoring mode. The method included a chromatographic run of 5.0 min using a C₁₈ analytical column and the calibration curve was linear over the concentration range of 0.5–2,000 ng mL⁻¹ with a correlation coefficient R of 0.998 or better. The intra- and inter-day precision ranged from 3.4 to 5.6% and 4.3 to 8.2%. The intra- and inter-day assay accuracy was between −7.4 and 8.6%. The method was successfully applied for the pharmacokinetic study on jasminoidin in rabbit after a single dose oral administration of Gardenia jasminoides Ellis (Gardenia) and Gardenia coupling Coptis chinensis Franch (Coptidis) extracts.

     

LC-UV Column-Switching for Quantitation of Vitamin K1 in Infant Formula

This study was undertaken in order to develop an analytical method for vitamin K₁ in infant formula. The content of vitamin K₁ was investigated by using a column-switching LC-UV method. A Certified Reference Material sample of infant formula containing 0.94 ± 0.04 mg kg^?1 of vitamin K₁ was extracted with hexane followed by enzymatic digestion of fat and precipitation of the fatty acids. The linearity of this method was calculated using five consecutive standard curves, and the coefficient of determination (r ²) was found to be 0.9995. The limit of detection (LOD) and the limit of quantitation (LOQ) was 3.31 and 11.12 μg L^?1, respectively. The accuracy of intra- and inter-day measurements was in the range from 96.67 to 108.67%, and the precision of intra- and inter-day measurements was less than 5.13%. The recoveries were 109.27 ± 5.92%, and the recoveries of inter-laboratory results were in the range from 97.59 ± 1.29 to 109.27 ± 5.92%. The newly developed method uses the optimum conditions required to determine the content of vitamin K₁ in infant formula.     

Iridoid Glycosides from Gardenia jasminoides Ellis

Three new iridoid glycosides, 4″‐O‐[(E)‐p‐coumaroyl]gentiobiosylgenipin ( 1 ), 6′‐O‐[(E)‐caffeoyl]deacetylasperulosidic acid methyl ester ( 2 ), and 6′‐O‐[(E)‐sinapoyl]gardoside ( 3 ), together with seven analogues, 4 – 10 , were isolated from the BuOH extract of the fruits of Gardenia jasminoides Ellis . Their structures were determined by means of spectroscopic analyses, including HR‐ESI‐MS, IR, and ¹H‐ and ¹³C‐NMR, and 2D experiments (COSY, HSQC, and HMBC), and comparison with known related compounds.     

Simultaneous Determination of Five Major Biologically Active Ingredients in Different Parts of Gardenia jasminoides Fruits by HPLC with Diode-Array Detection

A simple, sensitive and specific HPLC method for the simultaneous separation and determination of geniposide, geniposidic acid, chlorogenic acid, crocin 1 and 2 in fruits of Gardenia jasminoides Ellis was developed. Optimum chromatographic performance was obtained with a C₁₈ column and acetonitrile—0.1% aqueous trifluoroacetic acid (v/v) as mobile phase. Isolated peaks were detected at 240 nm for geniposidic acid and geniposide, 330 nm for chlorogenic acid and 440 nm for crocin 1 and 2, respectively. Comparison of spectra recorded with a diode-array detector during elution of peaks enabled determination of method specificity. Quantitative determinations on different parts of gardenia fruit demonstrated that all these compounds were abundant mainly in pericarps and pulps and only iridoid glycosides were also presented in sepals and seeds of the fruits.     

Determination of Lorazepam in Rabbit Plasma After Intranasal Administration by LC-MS

An LC-MS method was developed and validated to determine lorazepam in rabbit plasma. Chromatographic separation was performed on a C₁₈ column using methanol-150 nM sodium acetate (62.5:37.5, v/v) as the mobile phase at the flow rate of 0.2 mL min^?1. The retention times for lorazepam and diazepam (internal standard) were 6 and 10 min, respectively. Quantitative analysis was operated in selected ion monitoring (SIM) and positive ion mode using target ions at [M + H]⁺ m/z 284.9 for diazepam and [M + Na]⁺ m/z 342.9 for lorazepam, respectively. The lower limit of quantification (LLOQ) was 1.2 ng mL^?1 and a linear range of 1.2–150 ng mL^?1 with correlation coefficients (r ²) of 0.9968. The intra- and inter-day relative standard deviation was <5 and < 10%, respectively. The accuracy values were higher than 95%. The method is simple, sensitive and repeatable, and has been successfully applied to pharmacokinetics studies of lorazepam-loaded mocroemulsions after intranasal administration in rabbit.     

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.     

Determination of Free and Total Piperacillin–Tazobactam in Plasma by HPLC–MS–MS: An Adapted Method for Neonates

A sensitive and specific liquid chromatography electrospray ionization-tandem mass spectrometry method for determination of total and free piperacillin–tazobactam in human plasma has been developed and validated. Plasma deproteinization was achieved with Amicon^® Ultra-0.5 mL centrifugal filter device (Millipore, Bedford, USA). Chromatography was performed on a Capcell Pak C₁₈ MG column (ID 2 mm × 100 mm, 5 μm, Shiseido, Kyoto, Japan) with isocratic elution using a mobile phase containing water and acetonitrile with an addition of 0.02% of formic acid. Detection was achieved by an Applied Biosystems API 3000 triple quadrupole mass spectrometer (ABI-SCIEX, Toronto, Canada). Electrospray ionization (ESI) was used for ion production. The limits of quantification were 100 ng mL⁻¹ for piperacillin and 30 ng mL⁻¹ for tazobactam. The precision and accuracy for both intra- and inter-day determination of piperacillin ranged from 2.8 to 9.1% and from 94.9 to 104.4%. The precision and accuracy for intra- and inter-day determination of tazobactam ranged from 2.9 to 9.3% and from 88.9 to 99.8%. The precision and accuracy for intra- and inter-day determination of free piperacillin ranged from 4.4 to 14.7% and from 89.0 to 109.6%. The precision and accuracy for intra- and inter-day determination of free tazobactam ranged from 2.8 to 14.4% and from 93.9 to 108.0%. Fifty and 150 μL plasma were used for total and free piperacillin–tazobactam analysis, respectively. The validation results of this analytical method made it feasible for being used in a further pilot study of population pharmacokinetics of piperacillin–tazobactam in neonates.

     

LC�CMS Method for Determination of Amphotericin B in Rabbit Tears and Its Application to Ocular Pharmacokinetic Study

A rapid and sensitive LC?CMS?CMS method was developed for the quantification of amphotericin B in rabbit tears using natamycin as internal standard. The analyte and internal standard were extracted from the tear sample using a solid phase extraction method. Chromatographic separation was achieved on a Phenomenex Luna 3 ??m CN column (100 × 2 mm, 3 ??m) using 3.5 mM ammonium acetate (pH 4):methanol (10:90) as mobile phase. The assay was validated with a linear range of 0.1?C3.2 ??g mL^?1 for amphotericin B using 10 ??L of tear sample. The intra- and inter-day assay precision ranged from 2.49 to 4.37 and 2.17 to 5.59%, respectively, and intra- and inter-day assay accuracy was between from 0.27 to 3.32 and ?0.51 to 3.72%, respectively. The method was successfully applied to the pharmacokinetic studies of amphotericin B eye drops in rabbit tears.     

LC Assay of Eletriptan in Tablets and In Vitro Dissolution Studies

Eletriptan (ELT) is a new selective serotonin agonist approved for the treatment of acute migraine headaches. A simple and rapid liquid chromatographic method was developed and validated for the assay of ELT in tablets. Chromatography was carried out on a 250 mm × 4.6 mm C₁₈ column at 30 °C. Acetonitrile–15 mM triethylamine solution (adjusted to pH 7.0 using concentrated o-phosphoric acid) (60:40, v/v) mixture was used as mobile phase at 1.0 mL min^?1 flow rate and UV detector was set at 225 nm. A linear response (r ²  = 0.9999) was observed in the range of 0.1–1.6 μg mL^?1. The method showed good recoveries (100.08 %) and the RSD values for intra- and inter-day precision were 0.78–1.93 and 1.10–2.15%, respectively. The method can be used for quality control assays and in vitro dissolution studies of ELT in tablets.     

A new triterpenoid from the fruits of Gardenia jasminoides var. radicans Makino

A novel triterpenoid 3α,16β,23,24-tetrahydroxy-28-nor-ursane-12,17,19,21-tetraen (1) was isolated from the fruits of Gardenia jasminoides var. radicans Makino. The structure of the new compounds was elucidated on the basis of spectroscopic analysis including MS and NMR data. Compound 1 was in vitro tested for cytostatic activity on human throat cancer (Hep-2) cell line by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide method and showed mild anticancer activity with the IC₅₀ of 31.2 μM.     

Pharmacokinetic Study of ZS-1, a Targeted Peptide to NCI-H1299, in Rats Following Intravenous Administration

To investigate the pharmacokinetics of ZS-1 following intravenous injection in rats, ZS-1 was administered at doses of 20, 30 and 45 mg kg^?1, respectively. Blood samples were collected at 0.5, 3, 8, 12, 15, 20, 30, 40 and 45 min. ZS-1 in rat plasma was measured by LC. The limit of detection (LOD) was 0.02 μg mL^?1. The relative standard deviation (RSD) of intra- and inter-day precisions were <10%, and the accuracy of intra- and inter-day were >94%. The mean extraction recovery of ZS-1 was 86.1%. After intravenous injection at doses of 20, 30 and 45 mg kg^?1, the concentration–time curves of ZS-1 fitted well to one compartment model. Area under the concentration–time curves (AUC) increased with dose. Clearance rates (CL) and elimination half-lives (T _1/2) had no significant difference between different dose groups (P > 0.05). ZS-1 was stable in plasma after at 25 °C for 2, 4, 6 h, after three freeze–thaw cycles, after ?20 °C for a month, and after ?80 °C for 3 months. The accuracy of ZS-1 was between 96.8 and 106.9%. The results indicated there was no significant degradation. These data indicated that the method for analysis of ZS-1 was reliable and the pharmacokinetic data could guide dosing regimens to be tested in future clinical pharmacokinetic study.     

Simultaneous Estimation of Vinpocetine and Its Metabolite, Apovincaminic Acid, in Beagle Plasma by LC-MS-MS

A method was developed to determine vinpocetine and its metabolite, apovincaminic acid, in beagle plasma by LC-MS-MS. After protein precipitation with methanol, the supernatant of the sample was concentrated and injected into an Agilent Zorbax XDB-C₁₈ column. The sample was separated by a mobile phase consisting of acetonitrile and 0.2% formic acid solution, and the reading was determined on an Agilent 6410 Triple Quad Tandem mass spectrometer in multiple reaction monitoring mode with the following transitions: m/z 351.5 ?? 280.2/266.3 for vinpocetine, 323.2 ?? 236.1/280.2 for apovincaminic acid, and 411.2 ?? 191.1 for the internal standard. The intra- and inter-day variances were less than 15% (RSD%), and average recoveries were higher than 80%. The linearity ranges (LR) between 0.1 and 20.0 ng mL^?1 for vinpocetine (r ² = 0.9980) and between 1.0 and 200.0 ng mL^?1 for apovincaminic acid (r ² = 0.9995) were established. In summary, this method is sensitive, specific, and appropriate for in vivo study of various dosage forms of vinpocetine.     

HPLC–MS Analysis of Isoniazid in Dog Plasma

A simple, rapid, and sensitive liquid chromatography–mass spectrometric (LC–MS) method was developed and validated for the determination of isoniazid in dog plasma. Plasma samples were deproteined with methanol and separated on a C₁₈ column interfaced with a single quadrupole mass spectrometer, using 0.1% formic acid–acetonitrile (91:9 v/v) as mobile phase. Detection was performed by positive electrospray ionization with selected ion monitoring at m/z 138 for isoniazid and 152 for entecavir maleate internal standard. Linearity was obtained over the range of 25–5,000 ng mL^?1, with a lower limit of quantification of 25 ng mL^?1. The intra- and inter-day precision was less than 2.7% in terms of relative standard deviation. Accuracy, expressed as relative error, ranged from ?2.0 to 8.0%. Plasma samples were analysed within 5 min. The method was successfully applied to the evaluation of the pharmacokinetics of isoniazid in dog plasma.     

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.     

Determination of Phenolic Acids in Root Exudates of Allelopathic Rice by Solid Phase Extraction-Ion Chromatography with Conductivity Detection

A simple, effective, and green ion chromatography method with conductivity detection was developed for the determination of benzoic acid, cinnamic acid, syringic acid, and p-hydroxybenzoic acid in the root exudates of allelopathic rice. The analytes were well separated within 25 min in an anion exchange column (150 mm × 4.0 mm i.d., 5 µm particle size) with mixtures of 6.4 m mol L^?1 Na₂CO₃ and 2.0 m mol L^?1 NaHCO₃ as the mobile phase at a flow-rate of 0.7 mL min^?1. Detection limits of benzoic acid, cinnamic acid, syringic acid, and p-hydroxybenzoic acid were 0.05, 0.20, 0.50, and 0.05 µg mL^?1, respectively. Intra- and inter-day precision was ≤4.0% and 3.2%, respectively, and the intra- and inter-day accuracy, indicated by relative error, ranged from ?8.0% to 9.0%. The developed method was successfully used to determine phenolic acids in the root exudates of allelopathic rice. The average recoveries of the analytes were between 90.7% and 103.0%.     

An Effective High-Performance Liquid Chromatographic–Mass Spectrometric Assay for Catecholamines, as the N(O,S)-Ethoxycarbonyl Ethyl Esters, in Human Urine

The purpose of this study was to develop a simple and accurate analytical method for determination of norepinephrine, epinephrine, and dopamine in urine. The method involves liquid–liquid extraction then liquid chromatography–mass spectrometry (LC–MS). Alkyl chloroformate derivatives were prepared, as the N(O,S)-alkoxycarbonyl alkyl esters of the analytes, in the aqueous samples. The optimum derivatizing reagent for preparation of the N(O,S)-alkoxycarbonyl alkyl esters was chosen by comparing the efficiency of LC of the derivatized analytes after liquid–liquid extraction. The optimum conditions for liquid–liquid extraction from the aqueous matrix were pH 3.0, no salt, and diethyl ether as extraction solvent. Limits of detection (LOD) were 0.5 ng mL^?1 for dopamine and epinephrine and 0.1 ng mL^?1 for norepinephrine. Limits of quantification (LOQ) for urine samples were 1.0 ng mL^?1 for all three compounds. The precision of intra- and inter-day assays was 1.65–581 and 7.17–9.73% (relative standard deviation, RSD), respectively. The range of inaccuracy for intra- and inter-day assays was ?6.47 to 11.9% and ?7.5 to 7.76% (bias) at concentrations of 5 and 50 ng mL^?1, respectively.     

Analysis of Isopropyl 3-(3,4-Dihydroxyphenyl)-2-hydroxypropanoate, a Metabolite of Danshensu, from Salvia miltiorrhiza, in Rabbit Plasma

Isopropyl 3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate (IDHP), a metabolite of Danshensu, from Salvia miltiorrhiza, has been proved to have potential as a novel drug for regulation of vasomotor activity in small-resistance vascular circulation. In this presentation we report a new specific method for analysis of IDHP in rabbit plasma. Plasma samples were pretreated with 1.5% formic acid in acetonitrile to remove the protein, and the resulting supernatant was extracted with ethyl acetate. Chromatographic separation was achieved on a C₁₈ column with 15.0% acetonitrile in 0.3% aqueous formic acid (pH 2.2) as mobile phase. Multiple-reaction-mode ion-trap mass spectrometry was selected for accurate analysis of IDHP. The calibration plot was linear in the range 0.1–200.0 ng mL^?1 for plasma samples. The detection limit was 0.02 ng mL^?1. Intra-day and inter-day coefficients of variation were <13.0% and intra-day and inter-day accuracy was within ±8.0% of known concentrations. Finally, the method was used to investigate the pharmacokinetics of IDHP in rabbits; the results indicated IDHP was eliminated rapidly after oral administration.     

LC Determination of trans-3,5,3′,4′,5′-Pentamethoxystilbene in Rat Plasma

A simple LC method has been developed and validated to determine trans-3,5,3′,4′,5′-pentamethoxystilbene (PMS) in rat plasma. Chromatographic separation was achieved through gradient delivery of acetonitrile and water (1.5 mL min^?1) at 50 °C. PMS was quantified using UV detection at 320 nm. The standard curve ranged from 15 to 1,500 ng mL^?1. The intra- and inter-day precisions, in terms of CV, were all less than 10% while the inter-day and intra-day bias ranged from ?6.8 to 8.3%. The plasma PMS levels were monitored in Sprague-Dawley rats after drug administration. This simple LC method appears to be useful in the pharmacokinetic investigation of PMS.     

An Accurate Assay for Simultaneous Determination of Irinotecan and Its Active Metabolite SN-38 in Rat Plasma by LC with Fluorescence Detection

An accurate LC method was developed and validated for simultaneous determination of irinotecan (CPT-11) and its active metabolite SN-38 in rat plasma. Plasma samples were pretreated with 0.4 g mL^?1 sodium dodecyl sulfate to inactive the carboxylesterase and avoid the conversion of CPT-11 to SN-38. Chromatographic separation was achieved on a Diamaonsil C₁₈ column using acetonitrile–50 mM phosphate buffered solution (30:70, v/v) at pH 4.0 as the mobile phase with the flow rate of 1 mL min^?1. The linear quantitation ranges for CPT-11 and SN-38 were 5.05–3,030 and 3.15–315 ng mL^?1 with r ² > 0.99, respectively. The lower limit of quantification (LLOQ) was 2.33 ng mL^?1 for CPT-11 and 0.26 ng mL^?1 for SN-38 with intra- and inter-day relative standard deviation of <12% and the accuracy values of >90%. The method was proved to be accurate and sensitive enough and was successfully applied to a pharmacokinetic study of CPT-11 in rats.     

Validated Liquid–Liquid Extraction and LC–ESI–MS Method for the Determination of Melitracen in Human Plasma

Melitracen and the internal standard (I.S.), trifluoperazine, were extracted from plasma by a convenient liquid-liquid extraction. Chromatographic separation was performed on a Thermo Hypersil-Hypurity C₁₈ with the mobile phase consisting of 10 mM ammonium acetate–methanol–acetonitrile. A single-quadrupole mass spectrometer with an electrospray interface was operated in the selected-ion monitoring mode to detect the [M + H]⁺ ions at 292 m/z for melitracen and 408 m/z for trifluoperazine. The method was validated over 0.4–50.0 ng mL^?1 for melitracen. The recovery was 73.52–78.91%, and the lower limit of quantitation (LLOQ) detection was 0.4 ng mL^?1 for melitracen. The intra- and inter-day precision of the method at three concentrations were 2.96–7.76% with accuracy of 95.75–100.48%. Stability of compounds was established in a battery of stability studies. The bioequivalence of melitracen in the two formulations was evaluated in 18 healthy Chinese male volunteers with this assay. The described method showed acceptable precision, accuracy, linearity, stability, specificity and can be widely used for pharmacokinetic studies, and routine therapeutic drug monitoring.