HPLC Method For Simultaneous Estimation Research Articles

Validated HPLC method for simultaneous estimation of khellol glucoside, khellin and visnagin in Ammi visnaga L. fruits and pharmaceutical preparations

Tea bags including fruits of Ammi visnaga L. are used in Egypt as remedy for the treatment of kidney stones. Our study focuses on developing simple and rapid method utilising HPLC for quantitative estimation of khellol glucoside (KG), khellin (KH) and visnagin (VS) simultaneously. Their concentrations were determined in A. visnaga L. fruits at different developmental stages and in pharmaceutical formulations together with following up them during shelf life. Separation was accomplished using HPLC. Perfect resolution between KG, KH and VS was possible through using a mobile phase consisting of water:methanol:tetrahydrofuran (50:45:5, v/v/v). Peaks were detected at 245 nm. The suggested method for the determination of KG, KH and VS was successful in determining the analytes of interest without any interference of other compounds and matrix. All validation parameters were satisfactory and the procedure was relatively easy and fast as extracts are evaluated without previous steps of purification.

An Improved HPLC Method for Simultaneous Estimation of Isocurcumenol, Ar-Turmerone and <I>α</I>, <I>β</I>-Turmerone in Hexane Soluble Fraction of <I>Curcuma Longa</I> and Its Formulations

An Improved HPLC Method for Simultaneous Estimation of Isocurcumenol, Ar-Turmerone and <I>α</I>, <I>β</I>-Turmerone in Hexane Soluble Fraction of <I>Curcuma Longa</I> and Its Formulations

Development and Validation of HPLC Method for Simultaneous Estimation of Brimonidine Tartrate and Timolol Maleate in Bulk and Pharmaceutical Dosage Form

Brimonidine tartrate and Timolol Maleate are used in treatment of glaucoma by decreasing intra ocular pressure. A validated HPLC method was developed for the assay of them. The method was performed on BDS HYPERSIL Cyano column (250×4.6 mm, 5 μ) and the mobile phase consisted of Ammonium acetate (pH 5.0, 0.01 M) - Methanol (40:60, V/V) which pumped at a flow rate equals to 1.5 ml/min at ambient temperature. 20 μl of drugs sample solutions were monitored at two fixed wavelengths (lambda=254.0 nm for Brimonidine Tartrate and 300.0 nm for Timolol Maleate). The proposed method was validated in terms of linearity, accuracy, precision and limits of detection and quantitation according to ICH.

Development and Validation of New HPLC Method for Simultaneous Estimation of L-Lysine Hydrochloride and L-Carnitine L-Tartrate in Pharmaceutical Dosage Form

Development and Validation of New HPLC Method for Simultaneous Estimation of L-Lysine Hydrochloride and L-Carnitine L-Tartrate in Pharmaceutical Dosage Form

Development and Validation of HPLC Method for Simultaneous Estimation of Amoxycillin Trihydrate and Potassium Clavulanate in Pure and Marketed Tablet Dosage Form

A rapid, sensitive and specific RP-HPLC method involving UV detection was developed and validated for determination and quantification of Amoxycillin Trihydrate and Potassium Clavulanate. Chromatography was carried out on pre-packed Inertsil C18 ( 5µm, 250×4.6mm) column using filtered and degassed mixture of Methanol: Aqueous disodium hydrogen phosphate pH 5 as mobile phase at flow rate of 1.0 ml/min and effluent was monitored at 249 nm. The method was validated in terms of Linearity, precision, accuracy, specificity, limit of detection, limit of quantification as per the ICH guidelines.The assay was linear over the concentration range of amoxycillin trihydrate and potassium clavulanaterespectively.The accuracy and precision were found to be within the specified limits. The method does require only nine minutes as run time for analysis which proves its adoptability.

Development and validation of a reversed-phase HPLC method for simultaneous estimation of clotrimazole and beclomethasone dipropionate in lotion and cream dosage form

Background: The combination of Clotrimazole and Beclomethasone dipropionate is used as anti-fungal and anti-inflammatory for external use in the form of cream and lotion. Aim: A simple, specific, economic, precise, and accurate reversed-phase high performance liquid chromatographic method development for the simultaneous estimation of clotrimazole (CT) and beclomethasone dipropionate (BD) in lotion and cream formulations. Materials and Methods: The chromatographic separation was achieved on a Kromasil C18 (150 mm × 4.6 mm, 5 μm) analytical column. A mixture of acetonitrile-water (70:30, v/v) was used as the mobile phase, at a flow rate of 1 ml/min and detector wavelength at 254 nm. The validation of the proposed method was carried out for specificity, linearity, accuracy, precision, limit of detection, limit of quantitation, and system suitability test as per ICH guideline. Results: The retention time of CT and BD was found to be 5.4 and 4 min, respectively. The linear dynamic ranges were from 2-16 μg/ml and 80-640 μg/ml for BD and CT, respectively. Limit of detection and quantification for BD were 0.039 and 0.12 μg/ml, for CT 1.24 and 3.77 μg/ml, respectively. Conclusions: The developed method was validated and found to be simple, specific, accurate and precise and can be used for routine quality control analysis of titled drugs in combination in lotion and cream formulation.

Development and validation of the liquid chromatographic method for simultaneous estimation of metformin, pioglitazone, and glimepiride in pharmaceutical dosage forms

Introduction: A simple, precise, and accurate HPLC method for simultaneous estimation of metformin hydrochloride (MET), pioglitazone hydrochloride (PIO), and glimepiride (GLIMP) was developed and validated. Materials and Methods: Chromatographic separation of the drugs was performed by using a Phenomenex-ODS-3 (C-18) column (250 × 4.60 mm, 5 μm) with a mobile phase consisting of methanol:acetonitrile:15 mM potassium dihydrogen phosphate (pH 4) in the proportion of 40:35:25 (v/v) at a flow rate of 1 ml/min. Detection was carried out using a UV-SPD-10AVP detector at 240 nm. Results: The retention time for MET, PIO, and GLIMP were 2.85 ± 0.03 min, 4.52 ± 0.03 min, and 7.08 ± 0.02min, respectively. Parameters such as linearity (0.2–50 μg/ml for MET, 0.2-30 μg/ml for PIO, and GLIMP, respectively), precision (intra-day % RSD was 1.01-3.24 and inter-day % RSD was 1.54-4.09 for MET; intra-day % RSD was 1.03-2.09 and inter-day % RSD was 2.26-3.10 for PIO; and intra-day% RSD was 1.00-3.15 and inter-day % RSD was 1.58-3.07 for GLIMP), accuracy (99.66 ± 0.14 for MET, 98.46 ± 0.40 for PIO, and 98.62 ± 0.39 for GLIMP), specificity and robustness were calculated in accordance with ICH guidelines. Conclusions: The method was proved to be simple, rapid, precise, accurate, and cost effective.

Development and validation of HPLC method for simultaneous estimation of piperine and guggulsterones in compound Unani formulation (tablets) and a nanoreservoir system

An attempt has been made to develop and validate a simultaneous HPLC method for novel approach of drug release via oil-in-water (o/w) nanoemulsion formulation and Habb-e-Khardal Unani tablet containing piperine and guggul sterones E and Z as main ingredients. Nanoemulsion was prepared by titration method using sefsol-218 as an oily phase, cremophor-EL as a surfactant, transcutol as a co-surfactant and distilled water as an aqueous phase. The formulation was optimized on the basis of thermodynamic stability and dispersibilty test. The nanoformulation was evaluated for particle size, surface morphology, electrical conductivity and viscosity determination. The in vitro dissolution was carried out by dialysis bag method. Drugs were quantified using an HPLC method developed in-house with a C(18) column as stationary phase and acetonitrile and water as mobile phase at λ(max) of 240 nm. The optimized formulation showed higher drug release, lower droplet size and less viscosity as compared with the conventional Habb-e-Khardal Unani tablet. The present study illustrated the potential of nanoemulsion dosage form in improving biopharmaceutic performance of piperine and guggul sterone. The HPLC method was also found to be quite sufficient for the routine quality control of formulations containing piperine and guggul sterone E and Z as ingredients and also for in vitro drug release studies.

Development and Validation of Spectrophotometric and HPLC Method for the Simultaneous Estimation of Salbutamol Sulphate and Prednisolone in Tablet Dosage Form

Simple, accurate, and reproducible UV spectrophotometric and HPLC method for simultaneous estimation of salbutamol (SAL) and prednisolone (PRE) was developed in the present work. The first developed method was Simultaneous equation method, wavelength selected are 227 nm for salbutamol and 244 nm for prednisolone respectively. Linearity was observed in concentration range of 6-20μg/ml for salbutamol as well as for prednisolone. Second developed method was RP-HPLC method using Thermo C18 column (4.6 mm i.d × 250 mm) and acetonitrile: 0.025M potassium dihydrogen orthophosphate buffer (pH adjusted to 3.5 with orthophosphoric acid) in the ratio of 30:70% v/v as mobile phase. For HPLC method, linearity was observed in the concentration range of 20-100μg/ml for salbutamol as well as for prednisolone and drugs was subjected to oxidation, hydrolysis, and heat to apply stress condition for degradation studies. Results of analysis were validated stastically and by recovery studies.

Development and validation of a simple isocratic HPLC method for simultaneous estimation of phytosterols in Cissus quadrangularis

Cissus quadrangularis L. is a promising remedy prescribed in the ancient Ayurvedic literature for bone fracture healing properties. As this activity has been extensively investigated and well established, a range of formulations containing C. quadrangularis has been marketed. This work reports the development and validation of a reliable RP-HPLC method for the analysis of phytosterols in the various extracts of the plant. The proposed method utilizes a Cosmosil C8 column (250 ΄ 4.6 mm) with a compatible Phenomenex C8 guard column with isocratic elution of acetonitrile and water (95:5 v/v) at 25°. An effluent flow rate of 2 ml/min and UV detection at 202 nm was used for the analysis of phytosterols. The described method was linear in the range of 1–500 μg/ml, with excellent correlation coefficients. The precision, robustness and ruggedness values were also within the prescribed limits (less than 2%). The recovery values were within the range, which indicates that the accuracy of the analysis was good and that the interference of the matrix with the recovery of phytosterols was low. The phytosterols were found to be stable in a stock solution for 48 h (% RSD was below 2%) and no interfering extra peaks were observed under controlled stress conditions. The proposed method is simple, specific, precise, accurate, and reproducible and thus can be used for routine analysis of C. quadrangularis phytosterols in quality control laboratories.

Simple HPLC Method for Simultaneous Estimation of Fluconazole and Tinidazole in Combined Dose Tablet

A rapid and sensitive reversed-phase high-performance liquid chromatographic method is developed for simultaneous estimation of fluconazole, an orally active triazole anti-fungal agent, and tinidazole, which belongs to the group of 5-nitroimidazoles in combined dose tablet. Chromatographic separation was on an ODS Hypersil C(18) column using 0.05 M potassium dihydrogen phosphate buffer (pH 3.25, adjusted with orthophosphoric acid) and acetonitrile (82:18, v/v) as the mobile phase at a flow rate of 1.5 mL/min with detection at 210 nm. The asymmetry factors are 1.36 +/- 0.04 for fluconazole and 1.26 +/- 0.07 for tinidazole with a total run time of less than 7 min. The calibration curves were linear in the range 6-14 microg/mL for fluconazole and 80-190 microg/mL for tinidazole. The method was validated with respect to linearity, precision, accuracy, and specificity. The mean recovery for fluconazole and tinidazole is 99.65 +/- 0.84 and 99.34 +/- 0.70, respectively. The utility of the procedure is verified by its application to the market formulation that was subjected to various stressed conditions. Two potential degradation products of tinidazole on exposure to alkaline stressed condition are well-resolved. The method separated the two target drugs and degradation products well. No chromatographic interference is observed.

Development and validation of a reversed-phase HPLC method for simultaneous estimation of ambroxol hydrochloride and azithromycin in tablet dosage form

A simple, precise and accurate reversed-phase liquid chromatographic method has been developed for the simultaneous estimation of ambroxol hydrochloride and azithromycin in tablet formulations. The chromatographic separation was achieved on a Xterra RP18 (250 mm × 4.6 mm, 5 μm) analytical column. A Mixture of acetonitrile–dipotassium phosphate (30 mM) (50:50, v/v) (pH 9.0) was used as the mobile phase, at a flow rate of 1.7 ml/min and detector wavelength at 215 nm. The retention time of ambroxol and azithromycin was found to be 5.0 and 11.5 min, respectively. The validation of the proposed method was carried out for specificity, linearity, accuracy, precision, limit of detection, limit of quantitation and robustness. The linear dynamic ranges were from 30–180 to 250–1500 μg/ml for ambroxol hydrochloride and azithromycin, respectively. The percentage recovery obtained for ambroxol hydrochloride and azithromycin were 99.40 and 99.90%, respectively. Limit of detection and quantification for azithromycin were 0.8 and 2.3 μg/ml, for ambroxol hydrochloride 0.004 and 0.01 μg/ml, respectively. The developed method can be used for routine quality control analysis of titled drugs in combination in tablet formulation.

Quantitative estimation of itopride hydrochloride and rabeprazole sodium from capsule formulation

Two simple, accurate, economical and reproducible UV spectrophotometric methods and one HPLC method for simultaneous estimation of two component drug mixture of itopride hydrochloride and rabeprazole sodium from combined capsule dosage form have been developed. First developed method involves formation and solving of simultaneous equations using 265.2 nm and 290.8 nm as two wavelengths. Second method is based on two wavelength calculation, wavelengths selected for estimation of itopride hydrochloride was 278.0 nm and 298.8 nm and for rabeprazole sodium 253.6 nm and 275.2 nm. Developed HPLC method is a reverse phase chromatographic method using phenomenex C18 column and acetonitrile: phosphate buffer (35:65 v/v) pH 7.0 as mobile phase. All developed methods obey Beer's law in concentration range employed for respective methods. Results of analysis were validated statistically and by recovery studies.

A Validated RP – HPLC Method for Simultaneous Estimation of Cefixime and Cloxacillin in Tablets

This paper presents a RP‐HPLC method for the simultaneous estimation of cefixime and cloxacillin in tablets. The process was carried out on C18 column (5 μm, 25 cm × 4.6 mm, i.d) using phosphate buffer (pH 5.0), acetonitrile and methanol in the ratio 80:17:3 respectively as a mobile phase at a flow rate of 2mL/min. Wavelength was fixed at 225 nm. The retention time of cefixime and cloxacillin was found to be 5.657 and 6.200 min, respectively. The developed method is rapid and sensitive and it can be used for estimation of combination of these drugs in tablets.

Spectrophotometric and HPLC methods for simultaneous estimation of pseudoephedrine hydrochloride and loratadine from tablets

Two simple, accurate, economical and reproducible UV spectrophotometric and one HPLC method for simultaneous estimation of two-component drug mixture of pseudoephedrine hydrochloride and loratadine in combined tablet dosage form have been developed. The first developed method employs multiwavelength spectroscopy using seven mixed standards and 257.0 nm and 283.0 nm as two wavelengths for estimation. The second method involves first derivative spectroscopy using 308.6 nm and 263.0 nm as zero crossing points for pseudoephedrine hydrochloride and loratadine respectively. For both spectrophotometric methods, 0.2 M hydrochloric acid was used as solvent. Linearity was observed in concentration range of 0-40 mg/ml of loratadine and 0-800 mg/ml of pseudoephedrine hydrochloride. Developed HPLC method is reverse-phase chromatographic method using Inertsil C 18 column and methanol:ammonium acetate buffer in ratio of 80:20 pH 7.5 as mobile phase. Nimesulide was used as internal standard for HPLC method. For HPLC method, linearity was observed in concentration range of 0-200 mg/ml of loratadine and 100-2000 mg/ml of pseudoephedrine hydrochloride. Results of analysis were validated statistically and by recovery studies.