Method For Determination Of Diazepam Research Articles

Eco-friendly RP-HPLC method for determination of diazepam in coated tablet

A new eco-friendly RP-HPLC method for determining diazepam in coated tablets was developed and validated. The separation was achieved on a Lichrosper® 100 RP-18 (250 mm × 4 mm, 5 µm particle size) using the isocratic elution mode with a mobile phase composed of a mixture of water (H2O) and ethanol (EtOH) in a 40:60 (v/v %) ratio and a flow rate of 1.0 mL/min. The injection volume was 10 μL, and the detection wavelength was set at 254 nm. The column temperature was maintained at 35 ºC. The method was validated according to the ICH guideline, emphasizing selectivity/specificity, linearity, sensitivity, accuracy, precision, and robustness. The analytical method greenness score and Eco-scale approach were used for the method’s greenness assessment. The method was applied to determine the assay of diazepam in coated tablets and the uniformity of dosage units by content uniformity. Applying the method proposed in this study in the pharmaceutical industry is considered environmentally sustainable. It would bring benefits in terms of a safer working environment and reduction of toxic waste formation without compromising the reliability of the analytical results.

RP-HPLC Method development and Validation for Rapid estimation of Diazepam in Bulk and Pharmaceutical Dosage Form

The main objective of the study is to establish simple, precise and rapid RP-HPLC method for determination of diazepam and its decomposition products present in bulk and pharmaceutical dosage form. Various parameters that can potentially affect the analytical method were investigated and optimized. The compounds were well resolved in an isocratic method using the mobile phase composition of potassium dihydrogen phosphate buffer(0.05M) along with diluted o-phosphoric acid adjusted to pH 4.5: methanol (40:60v/v) at a flow rate of 1 ml/min using C18 (25cm x 0.46 cm) Hypersil BDS column. The detection was carried out at wavelength of 248nm.The retention time of diazepam was 3.3 min. The developed method was validated by evaluating various parameters such as linearity, precision, accuracy, robustness, specificity, limit of detection, and limit of quantification according to the international council for harmonization guidelines. The standard calibration curve was obtained in the concentration range of 20µg/ml for diazepam. The % of label claim was found out to be 93.66 for diazepam. Observation of all these parameters leads to the point that developed RP-HPLC method is accurate, precise, linear, specific and robust. It can be successfully adopted for routine quality control analysis of Diazepam in bulk and pharmaceutical dosage form without any interference.

Method Development and Validation of UV Spectro Photometric Method for Determination of Diazepam in its Pure and Pharmaceutical Dosage Form

Method Development and Validation of UV Spectro Photometric Method for Determination of Diazepam in its Pure and Pharmaceutical Dosage Form

Development of a simple and sensitive HPLC-MS/MS method for determination of diazepam in human plasma and its application to a bioequivalence study.

We developed a simple, sensitive, and effective ultra-performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS) method with an electrospray ionization (ESI) interface in multiple reaction monitoring (MRM) and positive ion modes to determine diazepam concentrations in human plasma using voriconazole as an internal standard (IS). Diazepam and IS were detected at transition 285.2→193.1 and 350.2→127.1, respectively. After liquid-liquid extraction (LLE) using 1.2 ml of ethyl acetate:n-hexane (80:20, v/v), diazepam and IS were eluted on a Phenomenex Cadenza CD-C18 column (150 × 3.0 mm, 3 µm) with an isocratic mobile phase (10 mM ammonium acetate in water:methanol [5:95, v/v]) at a flow rate of 0.4 mL/min. The peak retention time was 2.32 min for diazepam and 2.01 min for IS, respectively. The lower limit of quantitation (LLOQ) was 0.5 ng/mL (S/N > 10) using 50 µL of plasma, and no interferences were observed in chromatograms. Our analytical method was fully validated and successfully applied to a bioequivalence study of two formulations of diazepam in healthy Korean volunteers.

A new high-performance thin-layer chromatographic method for determination of diazepam in spiked blood samples

A high-performance thin-layer chromatographic method combined with densitometric measurement and UV scanning was used to optimize a new mobile phase consisting of hexane and ethyl acetate (7:3 v/v) for the estimation of diazepam in spiked human blood. Efficacy and ease of extraction with three solvents, acetonitrile, diethyl ether, and chloroform, each at three different pH conditions were studied. Validation parameters for diazepam were found to be linear in the range of 0.5–20 μg/spot with r = 0.998, limit of detection (LOD), 0.2 μg, limit of quantification (LOQ), 0.5 μg. Usefulness of the new mobile phase for the separation of six common benzodiazepines, namely, nitrazepam, clonazepam, lorazepam, chlordiazepoxide, alprazolam, and clozapine from diazepam, was proved through their retention factor and UV pattern.

Determination of Diazepam in Human Plasma by Solid-Phase Microextraction and Capillary Gas Chromatography-Mass Spectrometry

A simple, sensitive, and reproducible solid-phase microextraction and capillary gas chromatography-mass spectrometry (SPME-GC-MS) method for determination of diazepam in human plasma is described. The optimum conditions for the SPME procedure were as following: direct extraction mode with a polydimethylsiloxane (PDMS) fiber (100 μm film thickness), 250 μL of sample plasma matrix modified with a solution containing sodium chloride (10% weight by volume) and 4.25 mL of a phosphate buffer solution (0.1mol L−1, pH 6.9), extraction temperature 55°C under a magnetic stirring rate of 2500 rpm for 30 min, followed by the drug thermal desorption (250°C) in a GC injection port for 10 min. The limit of quantification of diazepam in plasma was 10.0 ng mL−1, with a coefficient of variation lower than 14.0% and linearity from 10.0 to 1000.0 ng mL−1, which allows diazepam analyses from sub to therapeutic levels.

Simplified method for simultaneous determination of diazepam and its metabolites in urine by thin-layer chromatography and direct densitometry

A direct densitometric method for determination of diazepam and its metabolites in urine was developed. The proposed procedure involves acid hydrolysis of urine specimens, thereby converting diazepam and its metabolites into benzophenones [2-methylamino-5-chlorobenzophenone (MACB) and 2-amino-5-chlorobenzophenone (ACB)]. It is followed by extraction with chloroform—isopropanol (3:1, v/v). The two benzophenones were separated on thin-layer chromatography plates using hexane—diethyl ether—acetic acid (80:10:10) as a mobile phase. Quantitation of the MACB and ACB spots was achieved by direct ultraviolet densitometry. The limit of detection was 0.5 μg per ml of urine for both benzophenones. The proposed method is simple, rapid, reproducible and has been found to be effective for direct determination of diazepam and its metabolites in urine.

Thin-layer chromatographic method for determination of diazepam and its major metabolite, N-desmethyldiazepam, in human serum

Thin-layer chromatographic method for determination of diazepam and its major metabolite, N-desmethyldiazepam, in human serum