scholarly journals A NOVEL REVERSE-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD FOR SIMULTANEOUS ESTIMATION OF ELLAGIC ACID, QUERCETIN, AND PIPERINE IN AYURVEDIC FORMULATIONS

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.

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):  
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.


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.


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.


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):  
Sv Saibaba ◽  
Shanmuga Pandiyan P

Objective: To develop and validate reverse phase-high performance liquid chromatographic method for estimation of olanzapine in bulk and tablet dosage form.Methods: Chromatographic analysis was performed on XTerra C18 (150×3.5 mm inner diameter, 5 μm) column using a mobile phase consisting of buffer (potassium dihydrogen phosphate) and methanol (45:55% v/v) with a flow rate of 0.6 ml/minutes. The detection was carried out at 247 nm.Results: The calibration curve of olanzapine was linear in the range of 30-70 μg/ml. The mean % assay of marketed formulation was found to be 100.2%, and % recovery was observed in the range of 98-102%. Relative standard deviation for the precision study was found <2%.Conclusion: The developed method is simple, precise and rapid, making it suitable for estimation of olanzapine in bulk and tablet dosage form.Keywords: Olanzapine, Reverse phase-high performance liquid chromatographic, Validation.


2007 ◽  
Vol 90 (1) ◽  
pp. 21-27 ◽  
Author(s):  
Ali M El-Shamy ◽  
Seham S El-Hawary ◽  
Mostafa E M Rateb

Abstract Parthenolide, a germacranolide-type sesquiterpene lactone, was estimated in Tanacetum parthenium (L.) cultivated in Egypt by using colorimetric, planar chromatographic, and high-performance liquid chromatographic (HPLC) methods. Parthenolide levels in the open-field herb and aseptically germinated shoots were also compared by using the HPLC method. Parthenolide was produced and estimated for the first time in the callus culture of the plant. In addition, 2 Egyptian market preparations were analyzed for their parthenolide content by using the HPLC method. The relative standard deviations were 0.093, 0.095, and 0.098% (n = 5,5,and 7, respectively), and the corresponding recoveries were 98.2, 98.9, and 99.4% for the colorimetric, planar chromatographic, and HPLC determinations, respectively.


2011 ◽  
Vol 8 (s1) ◽  
pp. S41-S46
Author(s):  
Prafulla Kumar Sahu ◽  
M. Mathrusri Annapurna ◽  
Dillipkumar Sahoo

This paper describes a high-performance liquid chromatographic method for simultaneous estimation of nabumetone and paracetamol in binary mixture. The method was based on RP-HPLC separation and quantitation of the two drugs on hypersil C-18 column (250 mm × 4.6 mm) using a mobile phase consisting of acetonitrile and 0.05% aqueous acetic acid (70:30v/v) at flow rate of 1 mL min-1. Quantitation was achieved with PDA detector at 238 nm based on peak area with linear calibration curves at concentration ranges 5-25 µg mL-1for both the drugs. Naproxen sodium was used as internal standard. The method has been successively applied to pharmaceutical formulation. No chromatographic interference from the tablet excipients was found. The method was validated in terms of precision, robustness, recovery and limits of detection and quantitation. The intra and inter-day precision and accuracy values were in the acceptance range as per ICH guidelines.


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