Efficient high performance liquid chromatograph/ultraviolet method for determination of diclofenac and 4′-hydroxydiclofenac in rat serum (original) (raw)
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Journal of chromatography, 1993
An assay using reversed-phase high-performance liquid chromatography with ultraviolet detection, at 278 nm, was developed to measure diclofenac in human plasma and urine at concentrations suitable for biopharmaceutical studies. Indomethacin was used as internal standard and separation was performed at 40 degrees C on a C18 Spherisorb column with acetonitrile-0.1 M sodium acetate (35:65, v/v) (pH 6.3) as mobile phase. The sample preparation is simple and rapid (extractionless), and the total run time is less than 5 min. The retention time is 2.8 min for diclofenac and 3.6 min for indomethacin. The detection limit is 0.2 microgram/ml using a 20-microliters loop.
This work describes a simple, sensitive, rapid and economical analytical procedure for direct spectrophotometric evaluation of diclofenac sodium (DS) using aqueous medium without using a chemical reagent. Parameters like time, temperature, acidic and basic conditions and interference by analgesic drugs were studied for a 5µg ml-1 solution of DS at 276 nm. Under optimized parameters, a linear working range of 0.1-30 µg ml-1 with regression coefficient of 0.9998 and lower detection limit of 0.01 µg ml-1 was obtained. The method was applied for DS contents in tablets, serum and urine samples.
Journal of Chromatography B: Biomedical Sciences and Applications, 2001
A novel high-performance liquid chromatographic (HPLC) method for the quantification of diclofenac in human plasma was set up. Samples, added with ibuprofen (used as internal standard) were purified by solid-phase extraction using Abselut Nexus cartridges (Varian) not requiring pre-conditioning. Drugs of interest were eluted directly into the autosampler vials and injected. The recovery of diclofenac was 92%, the analysis lasted
Turkish Journal of Pharmaceutical Sciences, 2016
A simple, rapid and reliable high performance liquid chromatography method (HPLC) with ultraviolet detection (UV) was developed and validated according to ICH guidelines, for quantitative analysis and therapeutic drug monitoring of diclofenac sodium (DS) in human plasma. Plasma samples (0.7 mL) were acid hydrolysis by 100 µL, 1 M hydrochloric acid. Analytes were concentrated from plasma by liquid-liquid extraction with 2 mL ethyl acetate by repeated twice, which allows to obtain good extraction yields (98.75%-99.32%). The separation was achieved by employing C18 analytical column (3.5 µm particle size, 150 mmx3.9 mm I.D.) under isocratic conditions using acetonitrile and NaH 2 PO 4 mixture (42.5:57.5, v/v) as mobile phase (pH: 3.16) flow rate of 1.5 mL/min. Naproxen (3 µg/mL) was used as an internal standard (IS). The DS and IS were detected at 281 nm and eluted at 2.6 and 6.2 min, respectively. Total run time was 7 min. Method showed linearity with very good determination coefficients (r 2 =0.999), over the concentration range of 50-1600 ng/mL. Limits of detection (LOD) and quantification (LOQ) were 8.95 ng/mL and 27.12 ng/mL, respectively. Intra-day precision and accuracy were between 0.93-5.27; 1.74-9.81, respectively. Inter-day precision and accuracy were between 2.71-6.64; 2.03-9.16, respectively. This method was successfully applied for determination of DS plasma concentrations during a pharmacokinetic study in healthy volunteers (n=12) after an oral administration of Voltaren ® 75 mg/tablet and remarkable variations in DS levels were observed. In our study, on the contrary to equivalent doses of DS, the observed significant differences in plasma levels of DS, on 2 nd , 4 th and 6 th hours, can be explained by pharmacokinetic differences, that arise from mainly polymorphisms of CYP2C9 and CYP3A4, which are major enzymes responsible for DS metabolism.
High-Throughput Ultra-Performance LC-MS-MS Method for Analysis of Diclofenac Sodium in Rabbit Plasma
Journal of Chromatographic Science, 2014
A new UPLC-MS-MS method was developed and validated for quantification of diclofenac sodium in rabbit plasma. Acetonitrile-based protein precipitation method was used to extract the drug from plasma samples. Chromatographic separation was carried out on Acquity UPLC w BEH phenyl C18 1.7 mm, 2.1 3 50 mm column. Drug elution was facilitated by using mobile phase containing acetonitrile (0.1% glacial acetic) and water (pH 3.5), in a ratio of 75 : 25, flowing at 0.2 mL/min. Molecular ions were generated by using the positive electrospray ionization mode (ESI 1) and analyzed on a triple-quadrupole mass spectrometer. The ionic transitions of diclofenac (m/z 296 > 214 and 249.9) and flufenamic acid (internal standard) (m/z 282.1 > 166.9 and 244) were measured in multiple reaction modes. Developed method is simple, quick, precise and accurate over a linearity range of 80-4,000 ng/mL. The lower limit of quantification (LLOQ) for diclofenac was 80 ng/mL. The percentage recoveries of diclofenac at three quality control samples were 54
Journal of Chromatography B: Biomedical Sciences and Applications, 1989
sodium, sodmm o-(2,6-dlchlorophenyl )ammophenylacetate (Voltaren 1, 1s a potent non-steroidal anti-inflammatory and analgesic drug, which has been used successfully for several years m the treatment of rheumatic deseases [l-3] This compound 1s metabolized m animals and humans to the corresponding mono-and dlhydroxy and conjugated derlvatlves [4,5] Several methods have been described for its determmatlon m body flulds, includmg gas chromatography with electron-capture detection [ 6,7], gas chromatography-mass spectrometry and high-performance llquld chromatography (HPLC) with VV detection [lo-21
Journal of Pharmaceutical and Biomedical Analysis, 2003
High-performance liquid chromatography (HPLC) was used to analyze microdialysis samples obtained in vivo from human subcutaneous adipose tissue after topical application of the nonsteroidal anti-inflammatory drug diclofenac. For the reliable determination of diclofenac two different detection principles were applied in two different laboratories. One HPLC method utilized UV-detection at 280 nm, the other one used selected reaction monitoring mass spectrometry (MS). The HPLC-UV and -MS methods offered low limits of quantification of 10 and 1 ng/ml and an accuracy between 94.0 Á/126.7 and 89.3 Á/110.9%, respectively. However, a comparison showed that the HPLC-UV method failed to determine diclofenac in biological matrices, as both false negative and positive values were found. HPLC-MS is clearly superior to HPLC-UV due to a much more selective detection, increased sensitivity and shorter run times. #
Journal of Chromatography B: Biomedical Sciences and Applications, 1996
We have modified and validated a capillary GC-MS method reported by Kadowaki et al. [J. Chromatogr., 308 (1984) 329] fi)r the determination of diclofenac in human plasma by using heptane rather than benzene as an extraction agent. In addition, acetone was added to the samples as a deproteination agent which increased the recovery of diclofenac. These revised processes allowed clean extraction and near-quantitative recovery of analyte (>95%). Separation was achieved on an HP-1 column with helium as carrier gas. The parent ion peaks of diclofenac (m/z 277) and the internal standard, 4'-methoxydiclofenac (m/z 307), were monitored by a mass-selective detector using the selected-ion monitoring mode. The linear range for the routine assay was from 5 to 2000 ng/ml, The detection and lower quantifiable limits were 0.2 and I ng/ml, respectively, with no interference from plasma. The within-day and between-day coefficients of variation for high and medium concentrations were less than 5% and were less than 13% fl)r lo~ concentrations (10 ng/ml). This GC-MS assay method has been used t\~r pharmacokinetic and drug interaction studies in humans.
Brazilian Journal of Pharmaceutical Sciences, 2013
A rapid, simple and low cost method was developed to determine diclofenac potassium (DP) in oral suspension, using a reverse-phase column (C8, 150 mm x 4.6 mm, 5 µm), mobile phase containing methanol/buffer phosphate (70:30 v/v, pH 2.5), at a flow rate of 1.0 mL/min, isocratic method, and ultraviolet detection at 275 nm. A linear response (r = 1.0000) was observed in the range of 10.0-50.0 µg/mL. Validation parameters such as linearity, specificity, precision, accuracy and robustness were evaluated. The method presented precision (repeatability: relative standard deviation = 1.21% and intermediate precision: between-analyst = 0.85%). The specificity of the assay was evaluated by exposure of diclofenac potassium under conditions of stress such as hydrolysis, photolysis, oxidation and high temperature. The method presented accuracy values between 98.28% and 101.95%. The results demonstrate the validity of the proposed method that allows determination of diclofenac potassium in oral su...