Development and validation of HPLC method for analysis of dexamethasone acetate in microemulsions

A simple, rapid, accurate and sensitive method was developed for quantitative analysis of dexamethasone acetate in microemulsions using high performance liquid chromatography (HPLC) with UV detection. The chromatography parameters were stainless steel Lichrospher 100 RP-18 column (250 mm x 4 mm i.d., 5 μm particle size), at 30 ± 2 ºC. The isocratic mobile phase was methanol:water (65:35; v/v) at a flow rate of at 1.0 mL.min-1. The determinations were performed using UV-Vis detector set at 239 nm. Samples were prepared with methanol and the volume injected was 20 μL. The analytical curve was linear (r² 0.9995) over a wide concentration range (2.0-30.0 μg.mL-1). The presence of components of the microemulsion did not interfere in the results of the analysis. The method showed adequate precision, with a relative standard deviation (RSD) smaller than 3%. The accuracy was analyzed by adding a standard drug and good recovery values were obtained for all drug concentrations used. The HPLC method developed in this study showed specificity and selectivity with linearity in the working range and good precision and accuracy, making it very suitable for quantification of dexamethasone in microemulsions. The analytical procedure is reliable and offers advantages in terms of speed and low cost of reagents.

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VALIDATION OF ASSAY PROCEDURES OF AFOBAZOLEIN MICROCAPSULES

International Journal of Pharmacy and Pharmaceutical Sciences ◽

10.22159/ijpps.2017v9i5.16684 ◽

2017 ◽

Vol 9 (5) ◽

pp. 278

Author(s):

Polkovnikova Yulia Aleksandrovna ◽

Slivkin Aleksei Ivanovich ◽

Halahakoon Amila Jeewantha

Keyword(s):

High Performance ◽

Analytical Procedure ◽

Ammonium Phosphate ◽

Hplc Method ◽

Good Precision ◽

Assay Procedure ◽

Analytical Curve ◽

Validation Criteria ◽

Relative Standard ◽

Sufficient Precision

Objective: The aim of this work was focused on a simple, rapid, accurate and sensitive method for quantitative analysis of afobazole in microcapsules (afb-m) using high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection.Methods: The analytical procedure was based on the fractionation characteristics of the afobazole in the chromatography column. The chromatography parameters «Zorbax extend-C18» column (150×2.1 mm i.d. 5 µm particle size), at 40 °C. The isocratic mobile phase was 5.0 M ammonium phosphate buffer (pH 7.0): acetonitrile (30:70; v/v) at a flow rate of at 1.0 ml. min-1. The determinations were performed using UV-Vis detector set at 220 nm.Results: An assay procedure for afb-m has been validated from indices, such as specificity, correctness, precision, and linearity. The registered retention time of the afobazole stock solution and the sample solution was 1.5 min. The determined accuracy of the method was in the range of 99.67%-100.67%. The analytical curve was linear (r2-0.9996) over a wide concentration range (50%-150%). The method showed sufficient precision, with a relative standard deviation (RSD) smaller than 2%.Conclusion: The HPLC method developed in this study showed specificity and selectivity with linearity in the working range and good precision and accuracy, making it very suitable for quantification of afb-m. The developed assay procedure of afb-m was meeting all the requirements of ICH validation criteria and can be applied for standardisation drug form afb-m.

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METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF THYMOL AND EUGENOL BY USING RP-HPLC IN PURE AND IN EMULGEL FORMULATION

INDIAN DRUGS ◽

10.53879/id.58.07.11895 ◽

2021 ◽

Vol 58 (07) ◽

pp. 59-65

Author(s):

Vinita C. Patole ◽

Shilpa P. Chaudhari ◽

Keyword(s):

High Performance ◽

Method Development ◽

Limit Of Detection ◽

Hplc Method ◽

Simultaneous Estimation ◽

Limit Of Quantification ◽

Relative Standard ◽

Peak Areas ◽

80 A 20 ◽

Development And Validation

An attempt was made to develop a simple, selective, rapid and precise high-performance liquid chromatography (HPLC) method for simultaneous estimation of thymol and eugenol. Analysis was performed on a C18 column with the mobile phase consisting of solvent %A (water) and solvent %B (acetonitrile) with the following gradient: 0–1 min, 80 % A, 20 % B; 1–7 min, 40 % A and 60 % B; 7–12 min, 10 % A and 90 % B; and 12–15min, 80 % A and 20 % B at a flow rate of 0.6 mL/min. The compounds were well separated on a Thermo Scientific Hypersil BDS RP C18 column (4.6 mm × 150 mm, dp = 5 µm) and ultraviolet detection at 280 nm. The retention times of eugenol and thymol were 10.5 min and 11.6 min, respectively. Validation of the proposed method was carried out according to the guidelines of the International Council on Harmonization (ICH). The linearity of the method is good for thymol and eugenol over the concentration range of 1–50 ppm, and the r 2 values were 0.9996 for both thymol and eugenol. The calculated limit of detection (LOD) value was 0.5ppm and the limit of quantification (LOQ) value was 1ppm for both the analytes. The intra and interday relative standard deviation (RSD) of the retention time and peak areas was less than 3 %.The established method was appropriate, and the two markers were well resolved, enabling efficient quantitative analysis of thymol and eugenol.

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A NEW HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE SEPARATION AND SIMULTANEOUS QUANTIFICATION OF EPTIFIBATIDE AND ITS IMPURITIES IN PHARMACEUTICAL INJECTION FORMULATION

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2021v13i2.39895 ◽

2021 ◽

pp. 165-172

Author(s):

K. SRI GIRIJA ◽

BIKSHAL BABU KASIMALA ◽

VENKATESWARA RAO ANNA

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

High Performance ◽

Degradation Products ◽

Ultra Violet ◽

Hplc Method ◽

Chromatography Method ◽

Standard Drug ◽

Pharmaceutical Dosage Forms ◽

Relative Standard

Objective: The objective of the present study is to develop a stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) method for qualitative and quantitative determination of Eptifibatide and its impurities in bulk and pharmaceutical dosage forms. Methods: The chromatographic separation was carried on Phenomenex Luna C18 column (250 mm×4.6 mm; 5µ id) as stationary phase, methanol and phosphate buffer at pH 6.4 in the ratio of 65:45 (v/v) as mobile phase at flow rate of 1.0 ml/min, Ultra Violet (UV) detection was carried at the wavelength of 236 nm and the analysis was completed with a run time of 15 min. Results: In the developed conditions, the retention time of Eptifibatide and its impurities 1 and 2 were found to be 3.35, 4.93 and 8.18 min, respectively. The method was validated for system suitability, range of analysis, precision, specificity, stability and robustness. Spiked recovery at 50%, 100% and 150% was carried for both standard and impurities and the acceptable % recovery of 98-102 was observed for Eptifibatide and both impurities studied and the % Relative standard deviation (RSD) in each spiked level was found to be less than 2. Stability tests were done through the exposure of the analyte solution to five different stress conditions i. e expose to 1N Hydrochloric acid (HCl), 1 N Sodium hydroxide (NaOH), 3% Hydrogen peroxide (H2O2), 80 °C temperature to UV radiation. In all the degradation conditions, standard drug Eptifibatide was detected along with both the impurities studied and the degradation products were successfully separated. In the formulation analysis, there is no other chromatographic detection of other impurities and formulation excipients. Conclusion: The developed method was found to be suitable for the quantification of Eptifibatide and can separate and analyse impurities 1 and 2.

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BIO-ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS DETERMINATION OF LEDIPASVIR AND SOFOSBUVIR DRUGS IN HUMAN PLASMA BY RP-HPLC METHOD

International Journal of Current Pharmaceutical Research ◽

10.22159/ijcpr.2018v10i3.27223 ◽

2018 ◽

Vol 10 (3) ◽

pp. 21 ◽

Cited By ~ 1

Author(s):

Buyya Shyam Sunder ◽

Ashutosh Kumar Mittal

Keyword(s):

Human Plasma ◽

High Performance ◽

Method Development ◽

Hplc Method ◽

Pharmacokinetic Studies ◽

Buffer Solution ◽

Stability Of Solution ◽

Drug Concentrations ◽

The Stability ◽

Development And Validation

Objective: A novel, sensitive and accurate high-performance liquid chromatography with ultraviolet/visible light detection (HPLC-UV/VIS) method for the quantification of ledipasvir and Sofosbuvir in plasma was developed and validated. Methods: The analytes were extracted by liquid-liquid extraction method and chromatograph using a mobile phase consisting of acetonitrile and buffer solution, Methanol and Acetonitrile in the ratio of 200:600:200 (v/v) using Oyster BDS RP-C18 column. The flow rate 1.0 ml/min and UV detection at 238 nm were employed. The retention time for Ledipasvir and Sofosbuvir was 4.61 and 9.09 min respectively. Linearity for ledipasvir and Sofosbuvir was found to be in the range of 250-2000 ng/ml for both drugs respectively. Intra-and inter-day precision was less than 2% coefficient of variation.Results: The method was validated as per the USFDA guidelines and the results were within the acceptance criteria for selectivity, sensitivity, linearity, precision, accuracy, recovery stability of the solution, the stability of solution in plasma and dilution integrity.Conclusion: Majority of the HPLC method should be useful for monitoring human plasma drug concentrations, and pharmacokinetic studies in patients diagnosed with the Ledipasvir and Sofosbuvir formulations.

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ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF CURCUMIN AND CYCLOSPORINE BY RP-HPLC

International Journal of Pharmacy and Pharmaceutical Sciences ◽

10.22159/ijpps.2019v11i2.28975 ◽

2018 ◽

pp. 26-33

Author(s):

Neha Desai ◽

Munira Momin ◽

Upasana Singh ◽

Tabassum Khan ◽

Atul Sherje

Keyword(s):

High Performance ◽

Dosage Form ◽

Method Development ◽

Hplc Method ◽

Simultaneous Estimation ◽

Rp Hplc ◽

Relative Standard ◽

Method Development And Validation ◽

Precision Study ◽

Development And Validation

Objective: The present work was undertaken with an aim to develop and validate a rapid reverse-phase high-performance liquid chromatography (RP-HPLC) method for the estimation of curcumin and cyclosporine in the capsule dosage form. Methods: The RP-HPLC method for the simultaneous estimation of curcumin and cyclosporine was developed using Agilent (Infinity 1260) HPLC system and Eclipse XDB-C18 (4.6 x 150 mm i.d., 5µ) stationary phase. The optimized mobile phase comprised of acetonitrile: water: methanol (50: 10: 40 v/v/v) pumped at a flow rate of 0.5 ml/min. Separation of drugs was achieved in an isocratic mode and elution was monitored using PDA detector at 214 nm. The method was validated as per ICH-Q2R1 guidelines. Results: Retention time of the curcumin and cyclosporine were found to be 3.073 min and 6.373 min with the correlation coefficient (R2) of 0.9993 and 0.998 respectively. The response of curcumin and cyclosporine was found linear in the concentration range of 8-48 μg/ml and 4-24 μg/ml respectively. The percent recovery values were found in the range of 97-103% indicating satisfactory accuracy of the method. The percent relative standard deviation (% RSD) values for the precision study was less than 2 which suggest that the method is precise. Conclusion: The proposed method was found accurate, precise and specific for the determination of curcumin and cyclosporine in bulk as well as in capsule dosage form. Thus, the present method can be used for routine analysis and quality control of curcumin and cyclosporine in bulk and capsule dosage form.

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Development and validation of an HPLC method for the determination of fluorouracil in polymeric nanoparticles

Brazilian Journal of Pharmaceutical Sciences ◽

10.1590/s1984-82502013000100013 ◽

2013 ◽

Vol 49 (1) ◽

pp. 117-126 ◽

Cited By ~ 14

Author(s):

Ana Cristina de Mattos ◽

Najeh Maissar Khalil ◽

Rubiana Mara Mainardes

Keyword(s):

Quantitative Analysis ◽

Flow Rate ◽

High Performance ◽

Polymeric Nanoparticles ◽

Hplc Method ◽

Array Detector ◽

Photodiode Array Detector ◽

Relative Standard ◽

Photodiode Array ◽

Development And Validation

The objective of this work was to develop and validate a rapid high performance liquid chromatography (HPLC) method for the quantitative analysis of fluorouracil (5-FU) in polymeric nanoparticles. Chromatographic analyses were performed on an RP C18 column with a mobile phase consisting of acetonitrile and water (10:90, v/v) at a flow rate of 1 mL/min. The 5-FU was detected and quantitated using a photodiode array detector at a wavelength of 265 nm. The method was shown to be specific and linear in the range of 0.1-10 µg/mL (r = 0.9997). The precision (intra- and inter-day) was demonstrated because the maximum relative standard deviation was 3.51%. The method is robust relative to changes in flow rate, column and temperature. The limits of detection and quantitation were 10.86 and 32.78 ng/mL, respectively. The method fulfilled the requirements for reliability and feasibility for application to the quantitative analysis of 5-FU in polymeric nanoparticles.

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DEVELOPMENT AND VALIDATION OF CARBAMAZEPINE PLASMA CONCENTRATIONS MEASUREMENT AND ITS APPLICATION ON EPILEPSY PATIENTS

International Journal of Pharmacy and Pharmaceutical Sciences ◽

10.22159/ijpps.2017v9i9.19402 ◽

2017 ◽

Vol 9 (9) ◽

pp. 87 ◽

Cited By ~ 1

Author(s):

Astri Budikayanti ◽

Chiswyta Chaliana ◽

Melva Louisa ◽

Rianto Setiabudy

Keyword(s):

Lower Limit ◽

High Performance ◽

Limit Of Detection ◽

Plasma Concentrations ◽

Internal Standard ◽

Hplc Method ◽

Limit Of Quantification ◽

Relative Standard ◽

Precision And Accuracy ◽

Development And Validation

Objective: To develop and validate high-performance liquid chromatography with photodiode array (HPLC-PDA) detector as a method for measuring carbamazepine plasma concentrations in epilepsy patients treated with monotherapy or polytherapy.Methods: Carbamazepine was extracted from epilepsy patients’ plasma through liquid-liquid extraction, using protein precipitation with chloroform. Analysis was performed using HPLC with Inertsil DS-4 C18 (4.6x150 mm), 5 μm particle size column. The optimal condition for separation was established in a mobile phase consisting of acetonitrile: water (50:50) at a flow rate of 1.0 ml/min, detected by PDA detector at 220 nm. Propylparaben was used as the internal standard. The retention time was 3.5 min.Results: Linearity was obtained over a concentration range of 0.5-16 μg/ml with r = 0.999. The method showed good intra-and inter-day precision and accuracy of more than 90% difference (% diff) and 95% relative standard deviation (RSD). Lower limit of quantification (LOQ) was 0.5 μg/ml and lower limit of detection (LOD) was 0.2 μg/ml with 100% accuracy and more than 90% precision. Recovery test was nearly 100%. Stability of carbamazepine plasma concentration in 3 epilepsy patients was measured on the first and third month of treatment, ranging between 83.5 to 98.7%. When used to compare carbamazepine as a monotherapy versus polytherapy, the method showed good selectivity.Conclusion: The present HPLC method was valid for measuring carbamazepine plasma concentrations in epilepsy patients treated with monotherapy or polytherapy. This method meets the standard in the EMEA guideline in terms of linearity, precision, and accuracy, also selectivity in epilepsy patients treated with polytherapy.

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Optimasi dan Validasi Metode Kromatografi Cair Kinerja Tinggi untuk Menetapkan Kadar Asam Klorogenat dalam Ekstrak Etanol Daun Yakon (Smallanthus sonchifolius (Poepp. & Endl.) H. Robinson)

ALCHEMY Jurnal Penelitian Kimia ◽

10.20961/alchemy.16.1.28952.66-76 ◽

2020 ◽

Vol 16 (1) ◽

pp. 66

Author(s):

Zuhelmi Aziz ◽

Liliek Nurhidayati ◽

Syamsudin Abdillah ◽

Nancy Dewi Yuliana ◽

Partomuan Simanjuntak

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

Formic Acid ◽

Chlorogenic Acid ◽

High Performance ◽

Biological Activities ◽

Hplc Method ◽

Good Precision ◽

Smallanthus Sonchifolius ◽

Relative Standard

Yakon merupakan tanaman yang dapat digunakan untuk pengobatan dan kebutuhan pangan. Salah satu kandungan zat berkhasiat dalam daun yakon adalah asam klorogenat. Asam klorogenat diketahui memiliki aktifitas sebagai antioksidan, antikanker dan antidiabetes. Penentuan kadar asam klorogenat dalam matriks yang kompleks diperlukan metode yang selektif dengan ketelitian dan ketepatan yang baik. Pada penelitian ini dilakukan optimasi dan validasi metode kromatografi cair kinerja tinggi (KCKT) fase balik untuk penetapan kadar asam klorogenat. Ekstrak dibuat secara ultrasonikasi menggunakan pelarut etanol 95%. Kondisi optimum diperoleh menggunakan fase gerak asam format 0,1% dalam asetonitril–asam format 0,1% dalam air (gradien); fase diam oktadesilsilan (C18) pada suhu 30 ºC dan detektor UV pada panjang gelombang 328 nm. Metode KCKT ini memberikan hasil yang memiliki ketelitian yang tinggi dengan simpangan baku relatif 0,79% dan ketepatan yang baik dengan perolehan kembali 97,50%. Kadar asam klorogenat yang diperoleh dalam ekstrak etanol 95% daun yakon sebesar 1,02%. Optimization and Validation of High Performance Liquid Chromatography Methods for Determination of Chlorogenic Acid Levels in Ethanol Extracts of Yakon Leaves (Smallanthus sonchifolius (Poepp. & Endl.) H. Robinson). Yacon is a plant that can be used for medication and food needs. One of the bioactive compounds of yacon leaves is chlorogenic acid. Chlorogenic acid has various biological activities, such as antioxidant, anticancer and antidiabetic activities. To determine the chlorogenic acid in such complex matrix, such selective methods with good precision and acuracy are required. In this study, the optimization and validation of reverse phase high performance liquid chromatography (HPLC) method for chlorogenic acid determination were performed. The extract was prepared by ultrasonication in 95% ethanol.The optimized condition for HPLC obtained was by using mobile phase 0.1% formic acid in acetonitrile – 0.1% formic acid in water with gradient elution, stationary phase octadesylsilane (C18) at 30 oC and UV detector at of 328 nm. The result showed that HPLC method had high precicion with relative standard deviation of 0.79% and high accuracy with recovery of 97.50%. The chlorogenic acid in the ethanol 95% extract of Yacon leaves was 1,02%.<div> </div>

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DEVELOPMENT AND VALIDATION OF A RP-HPLC METHOD FOR THE SIMULTANEOUS DETERMINATION OF CURCUMIN, PIPERINE AND CAMPHOR IN AN AYURVEDIC FORMULATION

International Journal of Pharmacy and Pharmaceutical Sciences ◽

10.22159/ijpps.2018v10i4.25216 ◽

2018 ◽

Vol 10 (4) ◽

pp. 115

Author(s):

Sultana Shaikh ◽

Vandana Jain

Keyword(s):

High Performance ◽

Quantitative Estimation ◽

High Performance Liquid Chromatographic ◽

Hplc Method ◽

Column Temperature ◽

Rp Hplc ◽

Relative Standard ◽

Precision Study ◽

Development And Validation ◽

Liquid Chromatographic

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.

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UV-HPLC METHOD DEVELOPMENT AND VALIDATION FOR QUANTIFICATION OF BESIFLOXACIN HYDROCHLORIDE

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2017.v10i5.17456 ◽

2017 ◽

Vol 10 (5) ◽

pp. 250 ◽

Cited By ~ 1

Author(s):

Bijit Saha

Keyword(s):

High Performance ◽

Method Development ◽

High Performance Liquid Chromatographic ◽

Hplc Method ◽

Good Precision ◽

Analyte Solution ◽

Bulk Drug ◽

Hplc Method Development ◽

Development And Validation ◽

Liquid Chromatographic

Objectives: The objective of the present investigation was to develope and validates a new, rapid, accurate high performance liquid chromatographic (HPLC) method for the quantification of Besifloxacin Hydrochloride.Methods: Isocratic UV-HPLC separation was performed using a Zodiac C18 (150 X 4.6 mm) column, with 150 volume of Acetonitrile and 350 mL of Methanol in 500 mL buffer as mobile phase at a flow rate of 2 mL/min and UV detection at 295nm..Results: The sample found stable for 24 hours in analyte solution and compatible with nylon filter. The Beer’s law plots were found to be linear over the concentration range 70% to 130% with a correlation coefficient (r2) 0.9999 in diluent, phosphate buffer and simulated tear media. The %RSD was found less than 2% shows good precision, acceptable accuracy and reproducibility of the new method for the determination of Besifloxacin Hydrochloride.Conclusion: The method was validated as per the ICH guidelines. The method is accurate and can be applied for qualitative analysis of Besifloxacin Hydrochloride in bulk drug and in formulation.Keywords: Besifloxacin hydrochloride, High performance liquid chromatographic, Stress testing, Validation, Linearity, Accuracy, Precision.

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