Quantification of furosemide in camel plasma by high resolution mass spectrometry, application on pharmacokinetics.

We developed and validated a high-resolution liquid chromatography mass spectrometry method for the quantification of furosemide in camel plasma which was used for a pharmacokinetic study in camels. Plasma samples were extracted by supported liquid extraction and furosemide and internal standard (furosemide-D5) were separated on a an Agilent Zorbax XDB C18 column (50 × 2.1 mm i.d., 3.5 µm). Data was acquired in full-scan mode over a mass range of 200-400 Da in negative electrospray mode at a resolution of 70,000. Linear calibration curves were obtained over the concentration ranges of 1.0-10,000 ng/mL. The validated method was then successfully applied in evaluating the pharmacokinetics and metabolites of furosemide in six camels (Camelus dromedarus) and we were able to advice on a withdrawal time of furosemide treatment before racing.

Pharmacokinetics and metabolism study of firocoxib in camels after intravenous administration by using high-resolution bench-top orbitrap mass spectrometry.

In this study, we developed a high-resolution liquid chromatography mass spectrometry method for the pharmacokinetic study of firocoxib followed by full method validation. Following a solid-phase extraction, the firocoxib and internal standard (celecoxib) were separated on an Agilent Zorbax ZDB C18 column (50 mm × 2.1 mm i.d., 3.5 µm) with a gradient elution using methanol and 0.1% aqueous formic acid. Data acquisition was performed at 25,000 resolution with the automatic gain set to 1,000,000 and the maximum injection time of 100 ms. Data were acquired in full-scan mode over a mass range of 100-550 Da in positive electrospray mode. Linear calibration curves were obtained over the concentration ranges of 0.5-200 ng/mL and no interfering peaks were detected at the retention time of firocoxib and internal standard in blank camel plasma samples. The mean extraction recoveries of firocoxib at three concentrations of 5, 25 and 75 ng/mL ranged from 92 to 104%. Coefficient of variation of intra-day and inter-day precision were both <10%. The accuracy of the method ranged from 95 to 107%. The validated method was then successfully applied in evaluating the pharmacokinetics and metabolism of firocoxib in camels (Camelus dromedarus) (n=5) following intravenous (i.v.) administration of a dose of 0.1 mgkg/body weight. The results obtained (mean ± SD) were as follows: the terminal elimination half-life (t1/2ß) was 5.75 ± 2.26 h, and total body clearance (ClT) was 354.1 ± 82.6 mL/kg/h. The volume of distribution at steady state (VSS) was 2344.4 ± 238.7 mL/kg. One metabolite of firocoxib was tentatively identified as desalkyl firocoxib (m/z 283). Firocoxib could be detected in plasma 3-5 days following i.v. administration in camels using a sensitive liquid chromatography high-resolution orbitrap mass spectrometry method.

The pharmacokinetics and metabolism of meloxicam in camels after intravenous administration.

The pharmacokinetics and metabolism of meloxicam was studied in camels (Camelus dromedarus) (n = 6) following intravenous (i.v.) administration of a dose of 0.6 mg·kg/body weight. The results obtained (mean ± SD) were as follows: the terminal elimination half-life (t(1/2ß) ) was 40.2 ± 16.8 h and total body clearance (Cl(T) ) was 1.94 ± 0.66 mL·kg/h. The volume of distribution at steady state (V(SS)) was 92.8 ± 13.7 mL/kg. One metabolite of meloxicam was tentatively identified as methylhydroxy meloxicam. Meloxicam and metabolite were excreted unconjugated in urine. Meloxicam could be detected in plasma 10 days following i.v. administration in camels using a sensitive liquid chromatography tandem mass spectrometry (LC/MS/MS) method.

Elimination and phase II metabolism of ethanol in camels after intravenous administration.

Ethanol elimination was studied in camels (n=8) after a single bolus intravenous dose of 0.1g/kg bodyweight (BW). Blood samples were then collected at set intervals. Ethanol and ethyl glucuronide (EtG) in blood were analysed by validated static headspace gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry (LC-MS) methods, respectively. Blood-ethanol concentration-time profiles were plotted for each camel and these were evaluated. A simple linear regression model was fitted to the selected data points and the slope of the fitted line was used to estimate the elimination rate, the distribution factor and turnover rate, which were 5.15 mg/dL blood/h, 0.55 L/kg and 0.028 g/h/kg, respectively. Blood EtG concentration-time profiles were also plotted for each camel. The elimination half-life of EtG, estimated by linear regression (using the values obtained after ethanol was completely eliminated) was 2.18 h. The theoretical initial blood concentration of EtG (C(0)), obtained by extrapolation to time zero was 23.4 µg/dL. The results will be useful in monitoring alcohol doping in camels using either parent drug or metabolite.

The disposition of theophylline in camels after intravenous administration.

The pharmacokinetics of theophylline were determined after an intravenous (i.v.) dose of 2.36 mg/kg in six camels and 4.72 mg/kg body weight in three camels. The data obtained (median and range) for the low and high dose, respectively, were as follows: the distribution half-lives (t1/2 alpha) were 1.37 (0.64-3.25) and 2.66 (0.83-3.5) h, the elimination half-lives (t1/2 beta) were 11.8 (8.25-14.9) and 10.4 (10.0-13.5) h, the steady state volumes of distribution (Vss) were 0.88 (0.62-1.54) and 0.76 (0.63-0.76) L/kg, volumes of the central compartment (Vc) were 0.41 (0.35-0.63) and 0.51 (0.36-0.52) L/kg, total body clearances (Clt) were 62.3 (39.4-97.0) and 50.2 (47.7-67.4) mL/h.kg body weight and renal clearance (Vr) for the low dose was 0.6 (0.42-0.96) mL/h.kg body weight. There was no significant difference in the pharmacokinetic parameters between the two doses. Theophylline protein binding at a concentration of 5 micrograms/mL was 32.2 +/- 3.3%. Caffeine was identified as a theophylline metabolite but its concentration in serum and urine was small. Based on the pharmacokinetic values obtained in this study, a dosage of 7.5 mg/kg body weight administered by i.v. injection at 12 h intervals can be recommended. This dosing regimen should achieve an average steady state serum concentration of 10 micrograms/mL with peak serum concentration not exceeding 15 micrograms/mL.

The activity of mixed function oxidases, estimated by in vivo antipyrine clearance, is similar in horses and camels.

The activity of hepatic mixed function oxidases was compared in horses and camels (Camelus dromedarius) by studying the pharmacokinetics of antipyrine in seven camels and five horses following intravenous administration of a single dose of antipyrine (25 mg/kg). The data obtained (mean +/- SEM and median in brackets) in camels and horses, respectively, were as follows: the elimination half-lives were 3.25 +/- 0.23 (3.19) and 3.09 +/- 0.25 (2.90) hr; the apparent volumes of distribution (area method) were 0.691 +/- 0.045 (0.648) and 0.642 +/- 0.034 (0.676) l/kg; the volumes of distribution at steady state were 0.659 +/- 0.040 (0.607) and 0.620 +/- 0.030 (0.653) l/kg; the volume of the central compartment of the two-compartment pharmacokinetic model were 0.386 +/- 0.0523 (0.349) and 0.298 +/- 0.05 (0.308) l/kg; total body clearances were 0.148 +/- 0.008 (0.158) and 0.145 +/- 0.007 (0.147) l/kg/hr; the areas under the curves to infinity were 171.0 +/- 9 (165) and 175 +/- 8.0 (170) micrograms.ml.hr. There was no statistical significance in any parameter between camels and horses which suggests that the activity of hepatic mixed function oxidases is similar in horses and camels.