Pharmacokinetics of methocarbamol and phenylbutazone in exercised Thoroughbred horses.

Methocarbamol (MCBL) is commonly used in performance horses for the treatment of skeletal muscle disorders. Current regulatory recommendations for show horses and racehorses are based on a single oral dose of 5 g, although doses in excess of this are often administered. The goal of the current study was to characterize the disposition of MCBL following higher dose administration and administration in combination with another commonly used drug in performance horses, phenylbutazone (PBZ). Exercised Thoroughbred horses were administered various doses of MCBL as a sole agent and MCBL in combination with PBZ. Blood samples were collected at various times, concentrations of MCBL and PBZ measured using LC-MS/MS and pharmacokinetic parameters calculated using compartmental analysis. Following administration of 15 g of MCBL, either as part of a single- or multiple-dose regimen, a number of horses exceeded the Association of Racing Commissioners International and the United States Equestrian Federation's recommended regulatory threshold at the recommended withdrawal time. There was not a significant difference between horses that received only MCBL and those that received MCBL and PBZ. Results of the current study support an extended withdrawal guideline when doses in excess of 5 g are administered.

Extraction of methocarbamol from human plasma with a polypyrrole/multiwalled carbon nanotubes composite decorated with magnetic nanoparticles as an adsorbent followed by electrospray ionization ion mobility spectrometry detection.

In this work, a polypyrrole/multiwalled carbon nanotubes composite decorated with Fe3 O4 nanoparticles was chemically synthesized and applied as a novel adsorbent for the extraction of methocarbamol from human plasma. Electrospray ionization ion mobility spectrometry was used for the determination of the analyte. The properties of the magnetic-modified adsorbent were characterized by scanning electron microscopy, transmission electron microscopy, Fourier transform IR spectroscopy, and X-ray diffraction. The effects of experimental parameters on the extraction efficiency of the sorbent were investigated. Under the optimized conditions, the linear dynamic range was found to be 2-150 ng/mL with the detection limit of 0.9 ng/mL. The relative standard deviation was 5.3% for three replicate measurements of methocarbamol in plasma sample. The extraction efficiency of the sorbent for the determination of different drugs with various polarities was also compared to that of Fe3 O4 -polypyrrole and Fe3 O4 -multiwalled carbon nanotubes sorbents. Finally, the method was used for the determination of methocarbamol in blood samples.

Simultaneous determination of methocarbamol and Ibuprofen by first derivative synchronous fluorescence spectroscopic method in their binary mixture and spiked human plasma.

Methocarbamol is formulated with Ibuprofen for treatment of alleviated pain associated with muscle spasm. This manuscript describes a sensitive and selective first derivative synchronous spectrofluorimetric method for simultaneous determination of both drugs. Factors affecting method selectivity were studied where best results were obtained upon using Δ λ = 20 and water as a solvent. Methocartbamol was determined at 283 nm while Ibuprofen at 285.5 nm in the concentration ranges of 0.4-5 and 0.2-4.8 µg/mL, respectively. The applicability of the proposed method was ascertained by application to different laboratory prepared mixtures and marketed formulation. The high sensitivity achieved by the proposed method permitted its application for determination of the drugs in human plasma spiked with pure drugs and their combined tablets. The proposed method showed no significant difference when compared with the reported HPLC method using student's t-test and F-ratio test.

The pharmacokinetics of methocarbamol and guaifenesin after single intravenous and multiple-dose oral administration of methocarbamol in the horse.

A simple LC/MSMS method has been developed and fully validated to determine concentrations and characterize the concentration vs. time course of methocarbamol (MCBL) and guaifenesin (GGE) in plasma after a single intravenous dose and multiple oral dose administrations of MCBL to conditioned Thoroughbred horses. The plasma concentration-time profiles for MCBL after a single intravenous dose of 15 mg/kg of MCBL were best described by a three-compartment model. Mean extrapolated peak (C0 ) plasma concentrations were 23.2 (± 5.93) µg/mL. Terminal half-life, volume of distribution at steady-state, mean residence time, and systemic clearance were characterized by a median (range) of 2.96 (2.46-4.71) h, 1.05 (0.943-1.21) L/kg, 1.98 (1.45-2.51) h, and 8.99 (6.68-10.8) mL/min/kg, respectively. Oral dose of MCBL was characterized by a median (range) terminal half-life, mean transit time, mean absorption time, and apparent oral clearance of 2.89 (2.21-4.88) h, 2.67 (1.80-2.87) h, 0.410 (0.350-0.770) h, and 16.5 (13.0-20) mL/min/kg. Bioavailability of orally administered MCBL was characterized by a median (range) of 54.4 (43.2-72.8)%. Guaifenesin plasma concentrations were below the limit of detection in all samples collected after the single intravenous dose of MCBL whereas they were detected for up to 24 h after the last dose of the multiple-dose oral regimen. This difference may be attributed to first-pass metabolism of MCBL to GGE after oral administration and may provide a means of differentiating the two routes of administration.

Rationally designed molecularly imprinted polymers for selective extraction of methocarbamol from human plasma.

Molecularly imprinted polymers (MIPs) with high selectivity toward methocarbamol have been computationally designed and synthesized based on the general non-covalent molecular imprinting approach. A virtual library consisting of 18 functional monomers was built and possible interactions between the template and functional monomers were investigated using a semiempirical approach. The monomers with the highest binding scores were then considered for additional calculations using a more accurate quantum mechanical (QM) calculation exploiting the density functional theory (DFT) at B3LYP/6-31G(d,p) level. The cosmo polarizable continuum model (CPCM) was also used to simulate the polymerization solvent. On the basis of computational results, acrylic acid (AA) and tetrahydrofuran (THF) were found to be the best choices of functional monomer and polymerization solvent, respectively. MIPs were then synthesized by the precipitation polymerization method and used as selective adsorbents to develop a molecularly imprinted solid-phase extraction (MISPE) procedure before quantitative analysis. After MISPE the drug could be determined either by differential pulse voltammetry (DPV), on a glassy carbon electrode modified with multiwalled-carbon nanotubes (GC/MWNT), or high performance chromatography (HPLC) with UV detection. A comparative study between MISPE-DPV and MISPE-HPLC-UV was performed. The MISPE-DPV was more sensitive but both techniques showed similar accuracy and precision.

Spectrofluorometric determination of methocarbamol in pharmaceutical preparations and human plasma.

A simple, sensitive and rapid spectrofluorometric method for determination of methocarbamol in pharmaceutical formulations and spiked human plasma has been developed. The proposed method is based on the measurement of the native fluorescence of methocarbamol in methanol at 313 nm after excitation at 277 nm. The relative fluorescence intensity-concentration plot was rectilinear over the range of 0.05-2.0 µg/mL, with good correlation (r=0.9999), limit of detection of 0.007 µg/ mL and a lower limit of quantification of 0.022 µg/ mL. The described method was successfully applied for the determination of methocarbamol in its tablets without interference from co-formulated drugs, such as aspirin, diclofenac, paracetamol and ibuprofen, The results obtained were in good agreement with those obtained using the official method (USP 30).The high sensitivity of the method allowed the determination of the studied drug in spiked human plasma with average percentage recovery of 99.42 ± 3.84.

Electrochemical determination of methocarbamol on a montmorillonite-Ca modified carbon paste electrode in formulation and human blood.

Utilizing the fascinating strong adsorptive ability, high chemical and mechanical stability properties of montmorillonite-calcium (MMT-Ca) clay, a MMT-Ca modified carbon paste electrode was developed for the sensitive determination of methocarbamol. Methocarbamol has been oxidized in buffered solutions at the developed MMT-Ca-modified CPE in a single 2-electron irreversible anodic peak. A simple and sensitive square-wave adsorptive anodic stripping voltammetric method was optimized for determination of methocarbamol in bulk form, pharmaceutical formulation and in spiked human serum using the developed modified CPE. This was carried out without the necessity for samples pretreatment and/or time-consuming liquid-liquid or solid-phase extraction steps prior to the analysis. The developed electrode exhibited an excellent sensitivity and selectivity towards methocarbamol even in the presence of 10(2)-10(4)-fold of its active ingredient "ibuprofen," common excipients, common metal ions or co-administrated drugs. Limits of detection of 3 x 10 (-9) and 1.2 x 10(-8) M and limits of quantitation of 1 x 10(-8), and 4 x 10(-8) M methocarbamol were achieved in the bulk form and in spiked human serum, respectively. Moreover, the developed method was successfully applied for determination of methocarbamol in human plasma of subjects following administration of an oral dose of Ibuflex tablets.

Determination of methocarbamol concentration in human plasma by high performance liquid chromatography-tandem mass spectrometry.

A simple and reproducible high performance liquid chromatography-tandem mass spectrometric method was developed for methocarbamol analysis in human plasma. Methocarbamol and the internal standard (IS) were extracted by a protein precipitation method. Under isocratic separation condition the chromatographic run time was 3.0 min. The calibration curve was linear over a range of 150-12,000 ng/mL with good intraday assay and interday assay precision (CV%<10.9%). The method was proven to be sensitive and selective for the analysis of methocarbamol in human plasma for bioequivalence study.

Determination of methocarbamol in equine serum and urine by high-performance liquid chromatography with ultraviolet detection and atmospheric pressure ionization-mass spectrometric confirmation.

Urine and serum samples collected from four standard-bred mares after and oral regimen administration of methocarbamol were extracted and analyzed. The method consisted of enzyme hydrolysis followed by a one-step liquid-liquid extraction, separation on a reversed-phase (RP-18) column, and detection using an ultraviolet (UV) detector. The confirmation was carried out using a liquid chromatography-atmospheric pressure ionization-mass spectrometry (LC-API-MS) system. Maximum methocarbamol concentrations of 1498, 1734, 1547, 2322 micrograms/mL in urine and 4.9, 1.7, and 3.6 micrograms/mL in serum were observed. The peak concentrations of the drug were detected 1-4 h (urine) and 10-60 min (serum) after administration to four horses. The method validation results and drug elimination profiles for both urine and serum are presented and discussed.

Development and validation of a high-performance liquid chromatographic method for the determination of methocarbamol in human plasma.

An isocratic HPLC method was developed and validated for the quantitation of methocarbamol in human plasma. Methocarbamol and internal standard in 200 microliters of human plasma were extracted with ethyl acetate, evaporated to dryness and reconstituted in water. Separation was achieved on a reversed-phase C18 column with a mobile phase of methanol-0.1 M potassium phosphate monobasic-water (35:10:55, v/v/v). The detection was by ultraviolet at 272 nm. Linearity was established at 1-100 micrograms/ml (r > 0.999). The limit of quantitation was designed as 1 microgram/ml to suit pharmacokinetic studies. Inter-day precision and accuracy of the calibration standards were 1.0 to 3.6% coefficients of variance (C.V.) and -2.0 to +1.6% relative error (R.E.). Quality controls of 3, 20 and 70 micrograms/ml showed inter-day precision and accuracy of 2.5 to 3.6% C.V. and -0.9 to -0.4% R.E. Recovery of methocarbamol was 91.4-100.3% in five different lots of plasma. The method was shown to be applicable on different brands of C18 columns.

High-performance liquid chromatographic analysis of methocarbamol enantiomers in biological fluids.

Methocarbamol enantiomers in rat and human plasma were quantified using a stereospecific high-performance liquid chromatographic method. Racemic methocarbamol and internal standard, (R)-(-)-flecainide, were isolated from plasma by a single-step extraction with ethyl acetate. After derivatization with the enantiomerically pure reagent (S)-(+)-1-(1-naphthyl)ethyl isocyanate, methocarbamol diastereomers and the (R)-flecainide derivative were separated on a normal-phase silica column with a mobile phase consisting of hexane-isopropanol (95:5, v/v) at a flow-rate of 1.6 ml/min. Ultraviolet detection was carried out at a wavelength of 280 nm. The resolution factor between the diastereomers was 2.1 (alpha = 1.24). An excellent linearity was observed between the methocarbamol diastereomers/internal standard derivative peak-area ratios and plasma concentrations, and the intra- and inter-day coefficients of variation were always < 9.8%. The lowest quantifiable concentration was 0.5 microgram/ml for each enantiomer (coefficients of variation of 9.8 and 8.8% for (S)- and (R)-methocarbamol, respectively), while the limit of detection (signal-to-noise ratio 3:1) was approximately 10 ng/ml. The assay was used to study the pharmacokinetics of methocarbamol enantiomers in a rat following intravenous administration of a 120 mg/kg dose of racemic methocarbamol and to evaluate plasma and urine concentrations in a human volunteer after oral administration of a 1000-mg dose of the racemate. The method is suitable for stereoselective pharmacokinetic studies in humans as well as in animal models.

Pharmacologic and pharmacokinetic properties of methocarbamol in the horse.

The hemodynamic, respiratory, and behavioral effects, as well as the pharmacokinetic properties of methocarbamol, were determined in horses. Heart rate, cardiac output, arterial and venous blood pressures, respiratory rate, and arterial blood gases did not change after IV methocarbamol (4.4, 8.8, 17.6 mg/kg) administration. There were no signs of behavior modification or ataxia observed. Analysis of plasma concentration time data indicated that the disposition of methocarbamol may be dose-dependent. Clearance and steady-state volume of distribution decreased as the dose increased. Plasma concentrations of guaifenesin, a metabolite of methocarbamol, were never greater than 0.5% of the plasma concentration of methocarbamol.