Archita J. Patel

Simultaneous spectrophotometric determination of celecoxib and diacerein in bulk and capsule by absorption correction method and chemometric methods.

Two methods, absorption correction and multivariate spectrophotometric methods were developed for simultaneous estimation of Celecoxib (CEL) and Diacerein (DIA) in combined dosage form. Absorption correction method involves direct estimation of DIA at wavelength 341 nm in which CEL has zero absorbance and shows no interference. For estimation of CEL, corrected absorbance was calculated at 253 nm due to the interference of DIA at this wavelength. Linearity was observed in the range of 6-22 μg mL(-1) for CEL and 3-11 μg mL(-1) for DIA. The method was validated as per ICH guidelines. Chemometric methods including classical least square (CLS), inverse least square (ILS), principal component regression (PCR) and partial least square (PLS) were studied for simultaneous determination of CEL and DIA in capsule using spectrophotometry. A set of 25 standard mixtures containing both drugs were prepared in range of 5-25 μg mL(-1) for CEL and 3-15 μg mL(-1) for DIA. Analytical figure of merit (FOM), such as sensitivity, selectivity, analytical sensitivity, limit of detection and limit of quantitation were determined for chemometric methods. The proposed methods were applied for determination of two components from combined dosage form.

Ultraviolet spectrophotometry (dual wavelength and chemometric) and high performance liquid chromatography for simultaneous estimation of meropenem and sulbactam sodium in pharmaceutical dosage form

UV spectrophotometric and high performance liquid chromatography (HPLC) methods were developed for simultaneous determination of meropenem (MERM) and sulbactam sodium (SB) in injection. UV spectrophotometric methods were developed using 0.1N sodium hydroxide as solvent. The Beers plot for dual wavelength method was linear in the range of 4-24 μg mL(-1) and 2-12 μg mL(-1) for MERM and SB, respectively. The percent recoveries were found to be 98.52±1.23% for MERM and 101.45±1.1% for SB. Chemometrics assisted UV spectrophotometry was performed using Partial Least Square (PLS) analysis model and Principal Component Regression (PCR) analysis model. The % recoveries of the MERM were found to be 100.61±0.06% and 101.31±0.12% using PLS and PCR, respectively. The % recoveries of the SB were found to be 98.29±0.09% and 97.61±0.13% using PLS and PCR, respectively. Chromatography was performed on Hypersil BDS C18 column using methanol:acetonitrile:water (10:20:70 v/v/v) as mobile phase. The retention times of MERM and SB were found to be 2.9 min and 2.25 min, respectively. Developed HPLC method was found to be linear in the range of 50-250 μg mL(-1) and 25-125 μg mL(-1) for MERM and SB, respectively. The % recoveries were found to be 98.85±0.25% and 98.63±0.34% for MERM and SB, respectively. The developed analytical methods did not show any interference of the excipients when applied to pharmaceutical dosage form.

SIMULTANEOUS HPTLC ANALYSIS OF ATORVASTATIN CALCIUM, EZETIMIBE, AND FENOFIBRATE IN TABLET

A simple, sensitive HPTLC method has been developed and validated for the simultaneous estimation of atorvastatin calcium, ezetimibe, and fenofibrate in tablet dosage form. Chromatographic separation was performed on aluminum plates precoated with silica gel 60 F254 with toluene:chloroform:methanol:glacial acetic acid (4.6:3:1.4:0.1, v/v/v/v) as mobile phase. The drugs were well resolved with RF values 0.20 ± 0.01, 0.33 ± 0.01, and 0.80 ± 0.02 for atorvastatin calcium, ezetimibe, and fenofibrate, respectively. Densitometric evaluation of the separated zones was performed at 253 nm. The method was validated in terms of linearity, accuracy, precision, and specificity. The calibration curve was found to be linear between 0.2 and 0.8 µg/spot for atorvastatin calcium (r2 = 0.999), 0.2 and 0.8 µg/spot for ezetimibe (r2 = 0.998), and 4 to 16 µg/spot for fenofibrate (r2 = 0.997). The limit of detection was found to be 0.019 µg/spot, 0.023 µg/spot, and 1.449 µg/spot for atorvastatin calcium, ezetimibe, and fenofibrate, respectively. The limit of quantitation was found to be 0.059 µg/spot, 0.068 µg/spot, and 4.390 µg/spot for atorvastatin calcium, ezetimibe, and fenofibrate, respectively. The proposed method can be used for estimation of the mentioned drugs in combined dosage form.

Stability-indicating high-performance thin-layer chromatographic method for simultaneous estimation of the active pharmaceutical ingredients metolazone and spironolactone

This work represents the validation of a stability-indicating thin-layer chromatographic technique for the simultaneous estimation of metolazone (METO) and spironolactone (SPIRO) from marketed formulation (tablets). Thin-layer chromatography was performed using precoated silica gel plate 60 F254 using ethyl ac-etate—chloroform—GAA (5:5:0.1 v/v) as the mobile phase for the separation of METO and SPIRO. The stability study forms an integral part of the formulation development process, and its use is also encouraged by various guidelines. Stress study was performed on active pharmaceutical ingredients (APIs) as well as on formulation for establishing a stability-indicating thin-layer chromatographic method for both drugs. The APIs were subjected to change under various environmental conditions such as pH, temperature, oxidation, etc. to determine their effect on the stability of drugs. The developed method was able to resolve drugs and their degradation products formed under the aforementioned conditions. The wavelength selected for quantitation was 238 nm. The method was validated as per the International Conference on Harmonization (ICH) guidelines and found to be linear in the range of 50–300 ng spot−1 for METO and 200–1200 ng spot−1 for SPIRO. The relative standard deviation (% RSD) values of the precision study were <2% which indicated that the developed method was precise; recovery was found to be 99.02–100.58% and 99.26–100.17% for METO and SPIRO, respectively. It could be concluded from the stability study that METO was prone to acidic hydrolysis and photolysis while SPIRO was prone to alkaline degradation.

Development and validation of stability indicating HPTLC method for simultaneous estimation of Fluocinolone acetonide and Miconazole nitrate in Ointment

Fluocinolone acetonide is a steroidal drug and Miconazole nitrate is an antifungal azole. The combination of these drugs has a highly beneficial effect on dermatological inflamatory disorders associated with fungal infections. A simple, specific, accurate, precise, robust and stability indicating HPTLC method has been developed and validated for the simultaneous estimation of Fluocinolone acetonide and Miconazole nitrate in bulk drug and ointment dosage form. Chromatography was performed using pre-coated silica gel aluminium plate 60F254, (10 ×10 cm) as stationary phase and n-Hexane: Ethyl acetate (1: 9, v/v) as mobile phase. Detection was carried out at 254 nm. The Rf value for Miconazole nitrate and Fluocinolone acetonide was found to be 0.46 and 0.64 respectively. The optimized conditions develop showed a linear response from 200–700 ng/spot (r2 =0.983) for Fluocinolone acetonide and 40000-140000 ng/spot (r2 =0.990) for Miconazole nitrate. The study involved observation on degradation products formed under different stress condition. The developed method successfully separated drug substances from degradation products formed under various stress conditions.

DEVELOPMENT AND VALIDATION OF REVERSE PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC AND HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHIC METHODS FOR SIMULTANEOUS ESTIMATION OF MELITRACEN HYDROCHLORIDE AND FLUPENTIXOL HYDROCHLORIDE IN BULK AND COMBINED DOSAGE FORM

Sensitive and reproducible reverse-phase high-performance liquid chromatographic and high-performance thin-layer chomatographic methods have been developed for simultaneous estimation of melitracen hydrochloride and flupentixol hydrochloride in bulk and combined tablet dosage form. The methods were validated according to the International Conference on Harmonization guidelines. In the RP-HPLC method, linearity was achieved with a detection range of 50–300 µg/mL (r2 = 0.998) and 2.5–15 µg/mL (r2 = 0.998) for melitracen hydrochloride and flupentixol hydrochloride, respectively. LOD was found to be 1.96 µg/mL and 0.032 µg/mL for melitracen hydrochloride and flupentixol hydrochloride, respectively. LOQ was found to be 5.96 µg/mL and 0.099 µg/mL for melitracen hydrochloride and flupentixol hydrochloride, respectively. In the HPTLC method, linearity was achieved with a detection range of 1600–6400 ng/band [800 µg/mL, 2–8 µ l] (r2 = 0.996) and 80–320 ng/band [40 µg/mL, 2–8 µ l] (r2 = 0.997) for melitracen hydrochloride and flupentixol hydrochloride, respectively. LOD was found to be 27.0 ng/band and 5.02 ng/band for melitracen hydrochloride and flupentixol hydrochloride, respectively. LOQ was found to be 81.81 ng/band and 13.60 ng/band for melitracen hydrochloride and flupentixol hydrochloride, respectively. The proposed methods can be used for estimation of the mentioned drugs in combined dosage form. The results obtained by applying the proposed methods were statistically analyzed.