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Development and validation of stability-indicating RP-HPLC method for the simultaneous determination of tizanidine HCL and meloxicam in rabbit's plasma

Zaman Muhammad, Hanif Muhammad, Khan Najm Ul Hassan, Mahmood Asif, Qaisar Muhammad Naeem, Ali Huma

Acta Chromatographica

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2019

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Vol. 31, no. 3

173--178

EN

High-performance liquid chromatography (HPLC) is a widely used technique for the simultaneous detection and quantification of different drugs. The purpose of the current study was to develop a simple and cost-effective reversed-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous determination of tizanidine (TZN) HCl and meloxicam (MLX) in rabbit's plasma. Assay of TZN and MLX was performed after extraction of drug from plasma by liquid–liquid extraction technique using methanol and diethyl ether as protein precipitants. Isocratic elution was performed in a Kromasil® C18 column (dimension, 250 × 4.60 mm; particle size, 5 μm) with mobile phase consisting of methanol–water (8:2). Orthophosphoric acid was used to adjust the pH of the mobile phase 3.0, and detection was done at 228 nm. Flow rate was 0.8 mL/min with ambient temperature and average operating pressure of 1400 psig. Retention time of TZN was 2.612 min and that of MLX was 6.960 min with a resolution of 3.18. Both drugs showed satisfactory linearity in the range of 10 to 50 ng/mL with correlation coefficients (R2) of 0.9989 and 0.9972 for TZN and MLX, respectively. The developed method was validated successfully for linearity, system suitability, intra-day and inter-day accuracy, and precision, robustness, and specificity following International Conference on Harmonization (ICH) guidelines. Conclusively, a precise, stable, reproducible, economical, and suitable method for estimation of pharmacokinetic evaluation was developed and validated.

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Development, validation, and pharmacokinetic application of liquid chromatographic method for estimation of raloxifene hydrochloride in rabbit plasma

Ravi P. R, Aditya N, Vats R

Acta Chromatographica

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2012

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Vol. 24, no. 4

559--573

EN

A rapid and sensitive reverse-phase liquid chromatographic method using ultraviolet detector was developed and validated for estimation of raloxifene hydrochloride in rabbit plasma. Plasma samples were extracted using simple protein precipitation-extraction method. The method was developed under isocratic conditions using a Zorbax SB-C8 analytical column with optimum mobile phase composition of 20 mM pH-4.5 ammonium acetate buffer-acetonitrile (63:37 υ/υ) at a flow rate of 1 mL min-1. The detector response was found to be linear in the concentration range of 50–1500 ng mL-1. High recoveries ranging from 97.2% to 100.2% were obtained, which precludes the use of internal standard. The developed method was found to be accurate, precise, and selective in the estimation of raloxifene hydrochloride in rabbit plasma based on the results of method validation carried out as per standard guidelines. The drug was found to be stable under various processing and storage conditions. The developed method was successfully applied in the estimation of raloxifene hydrochloride and the determination of various pharmacokinetic parameters post-intravenous bolus administration of drug in rabbits.

3

Development and validation of an RP-HPLC-UV method for the determination of ondansetron in rabbit plasma: Application to a pharmacokinetic study

Ravi S., Khan N., Darwis Y

Acta Chromatographica

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2011

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Vol. 23, no. 4

579--593

EN

A new sensitive and specific isocratic RP-HPLC-UV method was developed and validated for the determination of ondansetron in rabbit plasma using risperidone as an internal standard (IS). The sample preparation involved a simple deprotenization procedure with a mixture of 1 mL of acetonitrile and 50 μL of 10% w/υ zinc sulfate. Analysis was performed on a Phenomenex CN column (250 mm × 4.6 mm, 5 μm) with 50 mM ammonium acetate (pH 3.5) and acetonitrile (35:65, υ/υ) as mobile phase at a flow rate of 1.0 mL min-1. Column eluent was monitored at 310 nm. The calibration curve was linear over the concentration range of 25–1000 ng mL-1. (r2 = 0.9999) with a limit of quantification (LOQ) 25 ng mL. -1.The intraday and interday precision and accuracy were between 0.93% and 3.41% and −3.63% and 1.01%, respectively. The mean recoveries of ondansetron and risperidone were 85.87% and 99.80%, respectively. Ondansetroncontaining plasma samples were stable at −20°C for 14 days. The validated method was successfully applied for a pharmacokinetic study after a single oral administration of ondansetron (8 mg) to rabbits.