scholarly journals DEVELOPMENT AND VALIDATION OF NOVEL RP-HPLC METHOD FOR THE SIMULTANEOUS ESTIMATION OF ELLAGIC ACID AND QUERCETIN IN AN AYURVEDIC FORMULATION

2018 ◽  
Vol 10 (4) ◽  
pp. 111 ◽  
Author(s):  
Vandana Jain ◽  
Sultana Shaikh

Objective: To develop a novel, accurate, precise and linear reverse phase high performance liquid chromatographic (RP-HPLC) method for simultaneous qualitative and quantitative estimation of ellagic acid and quercetin in an ayurvedic formulation and validate as per international conference on harmonization (ICH) guidelines.Methods: In the present work, good chromatographic separation was achieved isocratically using a shim-pack HPLC C18 column (4.6 x 250 mm, 5μm) and a mobile phase consisting of 0.02 M potassium dihydrogen orthophosphate buffer (pH adjusted to 3.5 with orthophosphoric acid) and acetonitrile in the ratio 60:40, at flow rate of 1.2 ml/min and column temperature maintained at 35 °C. The effluents obtained were monitored at 255 nm with UV-visible detector.Results: The retention time of ellagic acid and quercetin were found to be 1.65 min and 2.94 min respectively. Linearity of ellagic acid and quercetin were tested in the range of 6-14 ppm and 3-11 ppm respectively. The correlation coefficient for ellagic acid and quercetin were 0.997 and 0.993 respectively. The high recovery values (98 %-102 %) indicate a satisfactory accuracy. The low percent relative standard deviation (% RSD) values in the precision study reveals that the method is precise.Conclusion: The developed method is novel, simple, precise, rapid, accurate and reproducible for simultaneous quantitative estimation of ellagic acid and quercetin in an ayurvedic formulation. Hence the developed method can be used for quantitative analysis and quality control of extracts and commercial samples of other plant species and formulation containing these two markers.

Author(s):  
Sultana Shaikh ◽  
Vandana Jain

Objective: The objective of the study was to develop a novel, accurate, precise, and linear reverse-phase high-performance liquid chromatographic (RP-HPLC) method for simultaneous qualitative and quantitative estimation of ellagic acid, quercetin, and piperine in different Ayurvedic formulations and validate as per the International Conference on Harmonization guidelines. Methods: In the present work, a good chromatographic separation was achieved isocratically using a shim-pack HPLC C18 column (4.6×250 mm, 5 μm) and a mobile phase consisting of 0.02 M potassium dihydrogen orthophosphate buffer (pH adjusted to 3.5 with orthophosphoric acid) and acetonitrile in the ratio 60:40, a flow rate of 1.2 ml/min and column temperature maintained at 35°C. The effluents obtained were monitored at 255 nm with ultraviolet-visible detector. Results: The retention time of ellagic acid, quercetin, and piperine was found to be 1.65 min, 2.94 min, and 14.57 min, respectively. The linearity of ellagic acid, quercetin, and piperine was tested in the range of 6–14 ppm, 3–11 ppm, and 3–13 ppm, respectively. The correlation coefficient for ellagic acid, quercetin, and piperine was found to be 0.997, 0.993, and 0.99, respectively. The high recovery values (98–102%) indicate a satisfactory accuracy. The low percent relative standard deviation values in the precision study reveal that the method is precise. Conclusion: The developed method is novel, simple, precise, rapid, accurate, and reproducible for simultaneous quantitative estimation of ellagic acid, quercetin, and piperine in Ayurvedic formulations. Hence, the developed method can be used for quantitative analysis and quality control of extracts and commercial samples of other plant species and formulations containing these three markers.


Author(s):  
Vandana Jain ◽  
Revati Sonone ◽  
Leena Tandel

The objective of this paper was to develop and validate a novel, simple, rapid, precise and accurate, reverse-phase high-performance liquid chromatographic (RP-HPLC) method for simultaneous quantitative estimation of piperine and embelin in the herbal formulation as per the International Conference on Harmonization guidelines (ICH). Chromatographic separation was achieved using a Cosmosil C-18 (250*4.6mm) SH 5.0 µm column with a mobile phase consisting of methanol and 0.02 M phosphate buffer in ratio 98:2 v/v, (pH adjusted to 2.3 with ortho-phosphoric acid) at a flow rate of 1 mL/min and column temperature maintained at 28°C and ultraviolet (UV) detection at 288 nm. The retention time of piperine and embelin was found to be 4.15 and 5.69 min respectively. The linearity of piperine and embelin was tested in the range of 5-40 ?g/mL. The correlation coefficient for piperine and embelin was found to be 0.997 and 0.995, respectively. The recovery values (98-102%) indicate a satisfactory accuracy. The method was found to be precise as the percentage relative standard deviation was found to be <2 %. The proposed novel isocratic RP-HPLC method is rapid (short run time below 10 min), precise, accurate and sensitive. The method was successfully applied for the simultaneous analysis of piperine and embelin in herbal formulation.


Author(s):  
Sultana Shaikh ◽  
Vandana Jain

Objective: To develop a novel, accurate, precise and linear reverse phase high performance liquid chromatographic (RP-HPLC) method for simultaneous qualitative and quantitative estimation of curcumin, piperine and camphor in an ayurvedic formulation and validate as per international conference on harmonization (ICH) guidelines.Methods: In the present work, good chromatographic separation was achieved isocratically using a shim-pack HPLC C18 column (4.6 x 250 mm, 5μm) and mobile phase consisting of 0.02 M potassium dihydrogen orthophosphate buffer (pH adjusted to 3.5 with orthophosphoric acid) and acetonitrile in the ratio 40:60, at flow rate of 1 ml/min and column temperature maintained at 35 °C. The effluents obtained were monitored at 255 nm with UV-visible detector.Results: The retention time of curcumin, piperine and camphor was found to be 6.57 min, 7.32 min and 8.57 min respectively. Linearity of curcumin and camphor were found in the range of 4-8 ppm and that of piperine was found to be 5-9 ppm. The correlation coefficient for curcumin, piperine and camphor were 0.998, 0.99 and 0.994 respectively. The high recovery values (98 %-102 %) indicate a satisfactory accuracy. The low percent relative standard deviation (% RSD) values in the precision study reveals that the method is precise.Conclusion: The developed method is novel, simple, precise, rapid, accurate and reproducible for simultaneous quantitative estimation of curcumin, piperine and camphor in an ayurvedic formulation. Hence the developed method can be used for quantitative analysis and quality control of extracts and commercial samples of other species containing these three markers.


Author(s):  
PRASANTHI CHENGALVA ◽  
LATHA LAVANYA PEDDAVENGARI ◽  
MADHAVI KUCHANA

Objective: The novel liposomal infusion formulation of cytarabine and daunorubicin liposomal infusion is considered as new hope in acute myeloid leukemia treatment. The objective of the present study is to develop and validate a simple, rapid, accurate, precise and sensitive reverse-phase high-performance liquid chromatographic (RP-HPLC) method for the simultaneous estimation of cytarabine and daunorubicin in bulk and infusion formulation. Methods: The chromatographic separation of the drugs was achieved on Denali C18 (250 mm×4.6 mm, 5 μm) in isocratic mode with mobile phase consisting of water (pH was adjusted to 3):acetonitrile in the ratio of 55:45 with a flow rate of 1 ml/min at a detection wavelength of 240 nm using photodiode array (PDA) detector. The column temperature was set at 30°C with 10 μl injection volume. The proposed method was validated as per the International council for Harmonisation (ICH) guidelines. Results: The retention times for cytarabine and daunorubicin were found to be 2.323±0.12 min and 3.140±0.16 min, respectively. Linearity (r2=0.999) was observed over a concentration range of 16.2–97.5 μg/ml for cytarabine and 7.2–43.5 μg/ml for daunorubicin. The percentage relative standard deviation (RSD) for precision studies was found to be 0.2 for both the drugs. Conclusion: A simple, rapid, economic, accurate, and precise RP-HPLC method was developed for simultaneous quantitative estimation of cytarabine and daunorubicin, and the method was validated as per the ICH guidelines. Hence, the method can be employed for the routine analysis of cytarabine and daunorubicin in bulk and infusion formulation.


2019 ◽  
Vol 16 (1) ◽  
pp. 12-23
Author(s):  
Nila Mary Varghese ◽  
Venkatachalam Senthil ◽  
Sajan Jose ◽  
Cinu Thomas ◽  
Jyoti Harindran

Objective: A simple, accurate and economical reverse phase- high performance liquid chromatographic method has been developed for the simultaneous quantitative estimation of two anti-retro viral drugs, Etravirine and Elvitegravir in nanoparticulate formulations for the first time. This method is a novel analytical technique for the detection of the both the drugs concurrently, as there is no method available for their simultaneous quantification, to the best of our knowledge. Methods: Optimization and validation of the chromatographic conditions were completed according to the standard ICH guidelines. The separation was done on a C18 column (250 mm x 4.6mm, 5μm) using methanol and phosphate buffer of pH (5.6) as the mobile phase in the ratio 78:22 v/v at a flow rate of 1ml/min for a short run time of 13 min. The detection wavelength was 285nm and the column temperature was maintained at 32oC. Results: The developed method was linear over 10 to 160 μg/ml with a regression coefficient of 0.999 for each. The LOD values were 4.83 and 9.25 µg/ml while LOQ values were 14.63 and 28.01 µg/ml for etravirine and elvitegravir respectively. The recovery values obtained by etravirine and elvitegravir were between 97.6% and 100.8%. Conclusion: The method was specific, precise, fast and accurate with good inter and intra day precision. The method was also effectively employed for the characterization and simultaneous quantification of both drugs in nanoparticulate formulation.


2020 ◽  
Vol 13 (3) ◽  
pp. 206-216
Author(s):  
Madhusudan Bhoir ◽  
Nutan Rao

The objective of the study was to develop and validate a novel, stability indicating, simple, rapid, accurate, precise and isocratic reverse-phase high-performance liquid chromatographic (RP-HPLC) method for simultaneous estimation of benserazide HCl and levodopa in a marketed formulation. Chromatographic separation was achieved by using C18 Cosmosil 4.6 × 250 mm column with a mixture of phosphate buffer pH 2 and acetonitrile in proportion of 95:5 as mobile phase at a flow rate of 1.0 ml/min and column temperature 25°C. The detection was carried out at 210 nm using UV detector. The retention time for benserazide and levodopa was found to be 3.1 minutes and 6.6 minutes respectively and recoveries from tablet were between 98 and 102 %.


Author(s):  
LEENA TANDEL ◽  
REVATI SONONE ◽  
VANDANA JAIN

Objective: The objective of this study was to develop and validate a novel, simple, rapid, accurate, and precise reverse-phase high-performance liquid chromatographic (RP-HPLC) method for simultaneous estimation of aloe-emodin and piperine in the ayurvedic formulation as per the International Conference on Harmonization guidelines. Methods: Chromatographic separation was achieved using a Prontosil C18 (250 × 4.6 mm, 5 μ), with a mobile phase consisting of 0.05% orthophosphoric acid and acetonitrile in the ratio of 50:50, at a flow rate of 1 ml/min and column temperature maintained at 28°C and ultraviolet (UV) detection at 225 nm. Results: The retention time of aloe-emodin and piperine was found to be 9.38±0.2 min and 13.45±0.2 min, respectively. The linearity of aloe-emodin and piperine was tested in the range of 1–20 μg/ml. The correlation coefficient for aloe-emodin and piperine was found to be 0.998 and 0.997, respectively. The recovery values (98–102%) indicate a satisfactory accuracy. The percentage relative standard deviation for precision was found to be <2% which indicates that the method is precise. Conclusion: A precise method for the simultaneous quantification of aloe-emodin and piperine was developed using high-performance liquid chromatography. Hence, the developed method can be used for quantitative and quality control analysis of formulations containing these phytoconstituents.


2017 ◽  
Vol 9 (6) ◽  
pp. 34
Author(s):  
Madhavi K. ◽  
Navamani M. ◽  
Prasanthi C.

Objective: To develop a simple, rapid, economic, accurate and precise reverse phase-high performance liquid chromatographic (RP-HPLC) method for the determination of hydrochlorothiazide and candesartan in the pharmaceutical dosage form and to validate as per international conference on harmonization (ICH) guidelines.Methods: The chromatographic separation was performed on Silanol BDS C18 column (250 x 4.6 mm, 5 μm), a mobile phase consisting of water (pH adjusted to 2.8 with orthophosphoric acid): acetonitrile (30:70 % v/v), with a flow rate 1 ml/min and the detection wavelength of 210 nm using photodiode array (PDA) detector.Results: The developed method resulted in elution of hydrochlorothiazide at 2.28 min and candesartan at 4.28 min. The calibration curves were linear (r2=0.999) in the concentration range of 6.25-18.75 μg/ml and 8-24 μg/ml for hydrochlorothiazide and candesartan respectively. The percentage recoveries were found to be 99.78-100.39 for hydrochlorothiazide and 99.87-100.64 for candesartan. The limit of detection (LOD) was found to be 0.410 μg/ml and 0.699 μg/ml for hydrochlorothiazide and candesartan respectively. The limit of quantitation (LOQ) was found to be 1.367 μg/ml and 2.330 μg/ml for hydrochlorothiazide and candesartan respectively.Conclusion: A simple, economic, accurate, precise, linear and rapid RP-HPLC method was developed for simultaneous quantitative estimation of hydrochlorothiazide and candesartan in bulk and pharmaceutical formulation and the method was validated as per ICH guidelines. Hence, the method holds good for the routine analysis of hydrochlorothiazide and candesartan in various pharmaceutical industries as well as in academics.


2018 ◽  
Vol 8 (6-s) ◽  
pp. 78-82 ◽  
Author(s):  
Vishal Singh Solanki ◽  
RAM SINGH BISHNOI ◽  
Raviraj Baghel ◽  
Deepti Jain

A simple precise and economical reverse phase high performance liquid chromatographic method has been developed and validated for the simultaneous estimation of Cilnidipine (CDP), Atenolol (ATL) and Chlorthalidone (CTD).The chromatographic separation was achieved by using Hypersil- keystone C18 (4.6 x 250mm, 5μm) under isocratic conditions The mobile phase consisted of methanol and triple distilled water (80/20, v/v) having pH 7 with a flow rate of 1.0 mL/min. The eluents were monitored at 225 nm for simultaneous measurement.The selected chromatographic conditions were found to effectively separate CDP (Rt: 3.25 min), ATL (Rt: 5.366 min) and CTD (Rt: 9.025 min) having good resolution. The developed method was validated for linearity, accuracy, precision, LOD, LOQ, robustness and for system suitability parameters as per ICH guidelines. In this study, an excellent linearity was obtained with r2 = 0.999, r² = 0.999, r² = 1, for CDP, ATL and CTD respectively. The developed chromatographic method proved to be simple, precise, accurate, robust and reproducible Thus, this method would be employed for routine simultaneous quantification of CDP, ATL and CTD in bulk form or tablet dosage form.   Keywords: Cilnidipine, Atenolol and Chlorthalidone, RP-HPLC.


2019 ◽  
Vol 9 (01) ◽  
pp. 83-88
Author(s):  
Pinkal Patel ◽  
Nalini Patel ◽  
Kinjal Parmar

A simple, selective and rapid reversed phase High Performance Liquid Chromatographic (RP-HPLC) method has been developed and validated for the simultaneous analysis Gatifloxacin and flurbiprofen sodium in eye drops. The separation was carried out using a mobile phase consisting ACN: Buffer (pH 3.5) in the ratio of 55:45 v/v. The column used was Phenomenex luna ODS C18 (250mm X 4.6 mm i.d., 5 μm particle size) with flow rate of 1 ml / min using UV detection at 268 nm. The described method was linear over a concentration range of 2-12 μg/ml for both of Gatifloxacin and flurbiprofen sodium. The retention times of Gatifloxacin and flurbiprofen sodium were found to be 3.710 min. and 6.797 min respectively. Method was validated statistically and recovery studies were carried out. The proposed method has been applied successfully to the analysis of cited drugs either in pure form or in pharmaceutical formulations with good accuracy and precision. The method here in described can be employed for quality control and routine analysis of drugs in pharmaceutical formulations.


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