HPTLC Method For Simultaneous Estimation Research Articles

Development and Validation of HPTLC Method for Simultaneous Estimation of Metolazone and Spironolactone in Bulk Drug and Pharmaceutical Dosage Form

A simple, precise and accurate HPTLC method has been developed for the simultaneous analysis of Metolazone and Spironolactone in bulk drugs and tablet formulation. Chromatographic separation was carried out on Merck HPTLC aluminium plates of Silica gel 60 F254, using n-propanol : triethylamine (7:3 v/v) as the mobile phase. HPTLC separation of the two drugs followed by densitometric measurement was performed in the absorbance mode at 240 nm. The drugs were resolved with R f values of 0.60 and 0.69 for Metolazone and Spironolactone respectively. The linear regression analysis data for the calibration plots showed good linear relationship with r value 0.99886 and 0.99862 for Metolazone and Spironolactone, respectively in the concentration range of 300 -700 ng/spot for Metolazone and 300-700 ng/spot for Spironolactone. The method was validated following the ICH guidelines in terms of accuracy, precision, specificity and robustness. The limit of detection and quantitation were 200 and 600 ng/spot respectively for Metolazone and 200 and 600 ng/spot for Spironolactone. The proposed developed and validated HPTLC method can be utilised for identification and quantitative analysis of Metolazone and Spironolactone in bulk drugs and pharmaceutical dosage form.

Validated Hptlc Method for Simulteneous Determination of Citicoline sodium and Piracetam in Combined Dosage Form

A simple, accurate and precise HPTLC method for simultaneous estimation of Citicoline sodium and Piracetam in a bulk and in combined dosages form is described in this paper. After optimization a mobile phase of Methanol: Water (16:4 v/v) was used during validation of an analytical method. Aluminum plate coated with the silica Gel 60 F254 was used as stationary phase. Densitometric evaluation of the separated bands was performed at wavelength 212 nm. The Rf values of Citicoline sodium and Piracetam were 0.30 and 0.68 respectively. The validated method was linear over the concentration range of 400 ng to 3200 ng /spot and 1200 ng to 3600 ng/spot of Citicoline sodium and Piracetam respectively. Precision of the method was evaluated by inerday and intraday RSD. The results were Citicoline sodium: Inter day RSD of peak response 0.62 % and Intraday RSD f peak response 1.05 % and for Piracetam : Interday RSD of peak response 1.44 % and Intraday RSD of peak response 1.06 %. Accuracy was determined in terms of percentage recovery at three concentration levels. The results are for Citicoline sodium: 97.66 %, 99.70 % and 97.91 % and for Piracetam: 98.52 %, 97.61 and 96.64 % respectively. Specificity was determined by spectral analysis of Citicoline sodium and Piracetam and overlaying the standard spectra and sample spectra respectively .There was no any interference of mobile phase and diluents at the Rf values of Citicoline sodium and Piracetam. Validation was done in accordance with the ICH Guidelines.

Validated Hptlc Method for Simulteneous Determination of Ceftriaxone Sodium and Sulbactam Sodium in Combined Dosage Form

A simple, accurate and precise HPTLC method for simultaneous estimation of Cefoperazone sodium and Sulbactam sodium as bulk and dry powder for injection in combined dosages form is described in this paper. A mobile phase of Chloroform: Ethyl alcohol: Diethyl amine: Water (14: 16:8:1.2 v/v) was used after optimization at different concentrations for the development of densitogram. Aluminum plate coated with the silica Gel 60 F254 was used as stationary phase. Densitometric evaluation of the separated bands was performed at 274 nm. The Rf values of Cefoperazone sodium and Sulbactam sodium were 0.41 ± 0.01 and 0.56 ± 0.01respectively. The validated method was linear over the concentration range of 200 ng to 900 ng /spot and 400 ng to 1800 ng/spot of Cefoperazone sodium and Sulbactam sodium respectively. Precision of the method was evaluated by inerday and intraday RSD. The results were Cefoperazone sodium: Inter day RSD of peak response 1.25 % and Intraday RSD 1.73 % and for Sulbactam sodium Interday RSD of peak response 1.54 % and Intraday RSD 0.98 %. Accuracy was determined in terms of percentage recovery at three concentration levels for Cefoperazone sodium RSD 99.20 %, 99.50 % and 100.32 % and for sulbactam sodium RSD 101.25 %, 100.40 % and 100.60 % respectively. Specificity was determined by spectral analysis of cefoperazone sodium and sulbactam sodium and overlaying the standard spectra and sample spectra respectively .There was no any interference of mobile phase and diluents at the RF values of Cefoperazone sodium and sulbactam sodium. Validation was done in accordance with the ICH Guidelines.

RAPID AND SENSITIVE SIMULTANEOUS ESTIMATION OF METFORMIN HYDROCHLORIDE AND PIOGLITAZONE HYDROCHLORIDE IN TABLET FORMULATION BY HPTLC METHOD

A rapid, simple, and sensitive HPTLC method for simultaneous estimation of two antidiabetic drugs, metformin hydrochloride and pioglitazone hydrochloride, in pharmaceutical tablet was developed and validated. Separation was achieved on aluminum sheets of silica gel 60 F254 using butanol:1, 4-dioxane:glacial acetic acid (5:3:2, v/v/v) as the mobile phase. This system gave a good resolution for metformin hydrochloride (R f value of 0.36 ± 0.01) and pioglitazone hydrochloride (R f value of 0.73 ± 0.01). Detection and quantification was carried out at 226 nm. Calibration plot was constructed in the range of 2000 to 20000 ng/band for metformin hydrochloride and 60 to 600 ng/band for pioglitazone hydrochloride. The linear regression data for the calibration plot showed a good relationship with r = 0.9960 and 0.9968 for metformin hydrochloride and pioglitazone hydrochloride, respectively. Assays for metformin hydrochloride and pioglitazone hydrochloride were 99.6% (RSD 0.618%) and 99.7% (RSD 0.511%), respectively, for the brand analyzed. The method was validated for precision and recovery. The limits of detection and quantification were 629.89 and 1908.76 ng/band for metformin hydrochloride and 8.51 and 25.77 ng/band for pioglitazone hydrochloride, respectively.

Stability-Indicating HPTLC Method for Simultaneous Estimation of Amoxicillin Trihydrate and Ambroxol Hydrochloride in Bulk and Pharmaceutical Dosage Form

A rapid, precise, selective and sensitive HPTLC method for simultaneous determination of Amoxicillin trihydrate (AMOX) and Ambroxol hydrochloride (AMBRO) in pharmaceutical dosage form was developed and validated in the present work. The chromatographic separation was performed on precoated silica gel 60 F254 plates with Ethyl acetate: methanol: toluene: water: glacial acetic acid (6.0: 3.0: 2.0: 1.0: 0.5 v/v) as the mobile phase with UV detection at 237nm. Retention factor for AMOX and AMBRO were found to be 0.32 ± 0.04 and 0.70 ± 0.05 respectively. Linearity was observed in the concentration range of 2000-12000ng/band for AMOX and 500-2500 ng/b and for AMBRO and the coefficient of regression for both the drugs was 0.9986 and 0.995 respectively. The method was validated for precision, robustness, and recovery and the values obtained were within the ICH limits. The LOD and LOQ values for AMOX were 105 and 220 ng per band respectively and for AMBRO 50 and 120 ng per band respectively. Drugs were subjected to oxidation, acid hydrolysis, base hydrolysis and sun light to apply stress condition for degradation studies as per ICH guidelines. The degradation products were well resolved from the pure drug with significantly different Rf values. Since the method could effectively separate the drug from its degradation products, it could be used as a stability-indicating method for analysis of individual drugs and the combined dosage form.

Validated HPTLC Method for Simultaneous Estimation of Atenolol and Aspirin in Bulk Drug and Formulation

This paper describes a new, simple, precise, and accurate HPTLC method for simultaneous estimation of Atenolol and Aspirin as the bulk drug and in tablet dosage forms. Chromatographic separation of the drugs was performed on aluminum plates precoated with silica gel 60 F254 as the stationary phase and the solvent system consisted of n-butanol : water : acetic acid (8 : 2 : 0.2 v/v/v). Densitometric evaluation of the separated zones was performed at 235 nm. The two drugs were satisfactorily resolved with values and for Atenolol and Aspirin, respectively. The accuracy and reliability of the method was assessed by evaluation of linearity (100–600 ng/spot for Atenolol and Aspirin), precision (intraday % RSD was 0.48–1.03 and interday % RSD was 0.68–1.14 for Atenolol, and intraday % RSD was 0.61–1.03 and interday % RSD was 0.69–1.04 for Aspirin), accuracy ( for Atenolol and for Aspirin), and specificity in accordance with ICH guidelines.

Validated HPTLC method for simultaneous estimation of ethinyl estradiol and drospirenone in bulk drug and formulation

This paper describes a new, simple, precise, and accurate HPTLC method for simultaneous estimation of ethinyl estradiol and drospirenone as bulk drug and in tablet dosage forms. Chromatographic separation of the drugs was performed on aluminum plates precoated with silica gel 60 F254 as the stationary phase and the solvent system consisted of toluene/methanol/ammonia (8:2:0.1) (v/v/v). Densitometric evaluation of the separated zones was performed at 280 nm. The two drugs were satisfactorily resolved with Rf values of 0.29±0.02 and 0.42±0.02 for ethinyl estradiol and drospirenone, respectively. The accuracy and reliability of the method was assessed by evaluation of linearity (150–400 ng/spot for ethinyl estradiol and 30–80 ng/spot for drospirenone), precision (intraday RSD 0.84–1.01% and interday RSD 0.76–1.04% for ethinyl estradiol, and intraday RSD 0.76–1.32% and interday RSD 0.73–1.01% for drospirenone), accuracy (98.48±0.21% for ethinyl estradiol and 98.30±0.15% for drospirenone), and specificity, in accordance with International Conference on Harmonisation (ICH) guidelines.

Development and Validation of HPTLC Method for Simultaneous Estimation of Gatifloxacin and Ornidazole in Human Plasma

A simple, rapid, sensitive and economic chromatographic method have been developed for simultaneous estimation of gatifloxacin and ornidazole in human plasma by using diclofenac sodium as internal standard The method employes a 20cm x10cm TLC plate precoated with silica gel 60F254 with 250μm thickness on aluminium sheets. The plasma sample was extracted using chloroform:formic acid (4.0:0.4, v/v). The mobile phase used consists of methanol: chloroform: ammonia solution [25%] (8.5: 1: 0.4 v/v/v), having chamber saturation for 20 min at room temperature. The R f values were found to be (R f =0.28±0.00178),(R f =0.77 ±0.00427) and(R f =0.50±0.00337) for GTFX, ORNZ and DICLO respectively. Densitometric analysis was carried out at wavelength 297.86 nm. The linear detector response was observed between 100 ng/band to 600 ng/band for both GTFX and ORNZ. The stability of gatifloxacin and ornidazol in plasma was confirmed during three freeze-thaw cycles (-20 IŠC), on bench during 12 hr and post preparative after 48 hr. The method was validated according to the FDA Bioanalytical guidelines with respect to selectivity, linearity,sensitivity precision and accuracy, recovery and stability.

Development and Validation of HPTLC method for simultaneous Estimation of Ciprofloxacin and Ornidazole in Tablet Dosage Forms

A simple, sensitive, precise and accurate HPTLC method has been developed for the simultaneous estimation of the ciprofloxacin and ornidazole in tablets. The stationary phase used was precoated silica gel 60F 254 plate. The mobile phase used was a mixture of n-butanol:ethanol:8M ammonia (5:0.5:1.5, v/v/ v). The densitometric scanning was carried out at 315 nm. This system was found to give compact spots for ciprofloxacin (R f value of 0.25 0.004) and ornidazole (R f value of 0.82 ± 0.007). The method was validated in terms of linearity, accuracy, precision, limit of detection, limit of quantification and specificity in accordance with the ICH guidelines. The calibration curve was found to be linear between 40 to 140 ng/spot for each ciprofloxacin and ornidazole with significantly high value of correlation coefficient (r 2 > 0.99). The limit of detection and quantitation were found to be 10.01 and 33.03 ng/spot, respectively for ciprofloxacin and the limit of detection and quantification were found to be 7.62 and 25.15 ng/spot, respectively for ornidazole. The results of analysis have been validated statistically and by recovery studies. The proposed HPTLC method can be successfully applied in the routine analysis of commercial pharmaceutical tablets.

Development and validation of a HPTLC method for simultaneous estimation of lornoxicam and thiocolchicoside in combined dosage form

Aim: To develop a simple, precise, rapid and accurate HPTLC method for the simultaneous estimation of Lornoxicam (LOR) and Thiocolchicoside (THIO) in bulk and pharmaceutical dosage forms. Materials and Methods: The separation of the active compounds from pharmaceutical dosage form was carried out using methanol:chloroform:water (9.6:0.2:0.2v/v/v) as the mobile phase and no immiscibility issues were found. The densitometric scanning was carried out at 377nm. The method was validated for linearity, accuracy, precision, LOD (Limit of Detection), LOQ (Limit of Quantification), robustness and specificity. Results: The Rf values (±SD) were found to be 0.84±0.05 for LOR and 0.58±0.05 for THIO. Linearity was obtained in the range of 60–360ng/band for LOR and 30–180ng/band for THIO with correlation coefficients r2=0.998 and 0.999, respectively. The percentage recovery for both the analytes was in the range of 98.7–101.2 %. Conclusion: The proposed method was optimized and validated as per the ICH guidelines.

A validated stability indicating HPTLC method for simultaneous estimation of irbesartan and hydrochlorothiazide

Introduction: Irbesartan, a diazaspiro angiotensin II blocker, is marketed in combination with Hydrochlorothiazide, which is a diuretic acting on distal convoluted tubule; for synergistic anti-hypertensive action. Methods: The present study deals with development and validation of a stability indicating HPTLC method for simultaneous estimation of Irbesartan and Hydrochlorothiazide using TLC plates precoated with Silica gel 60F254 and the mobile phase comprising Acetonitrile: Chloroform in the ratio of 5:6 v/v. Irbesartan and Hydrochlorothiazide were well resolved with Rf 0.27 ± 0.03 and 0.45 ± 0.03, respectively. Wavelength selected for the quantization was 270 nm. Inherent stability of these drugs was studied by exposing both drugs to various stress conditions as per ICH guidelines viz. Dry heat, oxidative, photolysis (UV and cool white fluorescent light) and hydrolytic conditions under different pH values. Results: Both the drugs were not degraded under dry heat and photolytic conditions, but showed degradation under hydrolytic condition. The degraded products of Irbesartan and hydrochlorothiazide were well resolved from the individual bulk drug response. Conclusion: The developed method is found to be simple, specific, precise and stability indicating. The specificity of the method was confirmed by peak purity profile of the resolved peaks.

Validated HPTLC Method for Simultaneous Estimation of Metformin Hydrochloride, Atorvastatin and Glimepiride in Bulk Drug and Formulation

This paper describes a new, simple, precise, and accurate HPTLC method for simultaneous estimation of Metformin hydrochloride (MET), Atorvastatin (ATV) and Glimepiride (GLM) as the bulk drug and in tablet dosage forms. Chromatographic separation of the drugs was performed on aluminum plates precoated with silica gel 60 F 254 as the stationary phase and the solvent system consisted of water: methanol: ammonium sulphate (1: 1: 4 v/v/v). Densitometric evaluation of the separated zones was performed at 237 nm. The three drugs were satisfactorily resolved with R F values 0.37 ± 0.02 and 0.59 ± 0.02, 0.75 ± 0.02 for MET, ATV, GLM respectively. The accuracy and reliability of the method was assessed by evaluation of linearity (200-700 ng/spot for MET, 600-2100 ng/spot for ATV and 600-2100 ng/spot for GLM ), precision (intra-day% RSD was 0.54–1.23 and inter-day% RSD was 0.90–1.48 for MET, intra-day% RSD was 0.91–1.74 and inter-day% RSD was 0.56–1.52 for ATV and intra-day% RSD was 0.60–1.27 and inter-day% RSD was 0.96–1.48 for GLM ), accuracy (99.66 ± 0.14 for MET, 98.46 ± 0.40 for ATV and 98.62 ± 0.39 for GLM ), and specificity in accordance with ICH guidelines.

A validated stability indicating HPTLC method for simultaneous estimation of irbesartan and hydrochlorothiazide

Introduction:Irbesartan, a diazaspiro angiotensin II blocker, is marketed in combination with Hydrochlorothiazide, which is a diuretic acting on distal convoluted tubule; for synergistic anti-hypertensive action. The present study deals with development and validation of a stability indicating HPTLC method for simultaneous estimation of Irbesartan and Hydrochlorothiazide using TLC plates precoated with Silica gel 60F254 and the mobile phase comprising Acetonitrile: Chloroform in the ratio of 5:6 v/v. Irbesartan and Hydrochlorothiazide were well resolved with Rf 0.27 ± 0.03 and 0.45 ± 0.03, respectively. Wavelength selected for the quantization was 270 nm. Inherent stability of these drugs was studied by exposing both drugs to various stress conditions as per ICH guidelines viz. Dry heat, oxidative, photolysis (UV and cool white fluorescent light) and hydrolytic conditions under different pH values.Results:Both the drugs were not degraded under dry heat and photolytic conditions, but showed degradation under hydrolytic condition. The degraded products of Irbesartan and hydrochlorothiazide were well resolved from the individual bulk drug response.Conclusion:The developed method is found to be simple, specific, precise and stability indicating. The specificity of the method was confirmed by peak purity profile of the resolved peaks.

Validated HPTLC Method for Simultaneous Estimation of Isotretinoin and Erythromycin in Bulk Drug and Topical Gel Form

A simple, precise and accurate high performance thin layer chromatographic method has been developed for the simultaneous estimation of Isotretinoin and Erythromycin in pharmaceutical gel. The separation was carried out on Merck TLC aluminum sheets of silica gel 60 F254, (20 × 10 cm) with 250 µm thickness using toluene: DMSO: methanol (6.5:0.2:2.5, v/v/v) as a mobile phase. HPTLC separation of the two drugs followed by densitometric measurement of their spots at 340 nm for Isotretinoin before derivatization and 410 nm for Erythromycin after derivatization with 10% H2SO4 and heating at 100°C for 15 min. The drugs were satisfactorily resolved with RF values of 0.38 ± 0.02 and 0.55 ± 0.02 for Isotretinoin and Erythromycin, respectively. The accuracy and reliability of the method was assessed by evaluation of linearity (30-150 ng spot-1 for Isotretinoin and 1200-6000 ng spot-1 for Erythromycin), precision (intra-day RSD 0.62-0.79% and inter-day RSD 0.43-0.71 % for Isotretinoin and intra-day RSD 0.47-1.71 % and inter-day RSD 0.42-1.49 % for Erythromycin), accuracy (98.91 ± 0.92 % for Isotretinoin and 99.27 ± 0.72 % for Erythromycin), and specificity, in accordance with ICH guidelines.

Development and validation of a method for simultaneous densitometric analysis of simvastatin and ezetimibe as the bulk drugs and in the tablet dosage form

Summary Simvastatin is a selective HMG-CoA reductase inhibitor and ezetimibe has lipid-lowering activity. Both are potential anti-lipidemic agents used in combination to reduce the amount of cholesterol and triglycerides in systemic circulation. This paper describes a simple, precise, and accurate HPTLC method for simultaneous estimation of the compounds as the bulk drugs and in the tablet dosage form. Chromatographic separation was performed on aluminium-backed silica gel 60 F254 plates with 8:2 (v/v) toluene-2-propanol as mobile phase. The separated spots were densitometrically evaluated at 240 nm. The drugs were satisfactorily resolved with RF values 0.48 ± 0.01 and 0.53 ± 0.01 for simvastatin and ezetimibe, respectively. The accuracy and reliability of the method were assessed by determination of validation data for linearity (0.4–2.0 μg per spot for both simvastatin and ezetimibe), precision (intra-day RSD 0.51–1.04%, inter-day RSD 0.34–1.11% for simvastatin; intra-day RSD 0.47–0.61%, inter-day RSD 0...