VALIDATION OF ASSAY PROCEDURES OF AFOBAZOLEIN MICROCAPSULES

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 A Simple HPLC-UV Method For the Quantification of Andrographolide in Self-Nano Emulsifying Drug Delivery System (Snedds) For Dissolution Study

International Journal of Drug Delivery Technology ◽

10.25258/ijddt.v7i04.10646 ◽

2017 ◽

Vol 7 (04) ◽

Author(s):

Syukri Y ◽

Afetma D. W. ◽

Sirin M. ◽

Fajri R. ◽

Ningrum A. D. K. ◽

...

Keyword(s):

Drug Delivery ◽

Drug Delivery System ◽

Delivery System ◽

Gastric Fluid ◽

Hplc Method ◽

Good Precision ◽

Intestinal Fluid ◽

Dissolution Medium ◽

Relative Standard ◽

Sufficient Precision

This research aim to validation of a simple, rapid and accurate HPLC-UV method for the quantification of andrographolide isolated from Andrographis paniculata Ness in Self Nano Emulsifying Drug Delivery System (SNEDDS) formulation during the dissolution test. The assay was performed using a XTerra® MS C18 column (150 mm X 4.6 mm, five μm) with a mobile phase of methanol and water (70: 30), at 0.8 mL/min flow rate and UV detection of 229 nm. Simulation gastric fluid (SGF) and intestinal fluid (SIF) were prepared as dissolution medium. The validation parameter was conducted including the test on linearity, precision, accuracy, LOD, and LOQ. The result showed an excellent linearity with r = 0.999 and good selectivity for both medium dissolution. The method showed sufficient precision, with a relative standard deviation (RSD) smaller than % Horwitz. The accuracy reported as % recovery was found to be 102.61 and 101.17 % in each SGF and SIF dissolution medium. LOD and LOQ were found 0.46 and 1.40 in SGF medium, 0.87 and 2.64 in SIF medium. In conclusion, the HPLC method developed showed specificity and selectivity with linearity in the working range, good precision and accuracy and suitable for quantification andrographolide in SNEDDS formulation.

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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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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.16.1.28952.67-76 ◽

2020 ◽

Vol 16 (1) ◽

pp. 67

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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METHOD VALIDATION OF RIFAMPICIN ANALYSIS IN HUMAN PLASMA AND ITS APPLICATION IN BIOEQUIVALENCE STUDY

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2019v11i4.33107 ◽

2019 ◽

pp. 296-303

Author(s):

ENDANG LUKITANINGSIH ◽

FATHUL JANNAH ◽

RATNA BUDHI PEBRIANA ◽

RATNA DEWI PUSPITA ◽

TAUFIQUROHMAN . ◽

...

Keyword(s):

Mobile Phase ◽

High Performance ◽

Bioequivalence Study ◽

Hplc Method ◽

Coefficient Of Determination ◽

European Medicines Agency ◽

Pharmacokinetic Profiles ◽

Relative Standard ◽

Precision And Accuracy ◽

Bioequivalence Test

Objective: This research aims to validate the method for rifampicin analysis in plasma by using High-Performance Liquid Chromatography (HPLC) that can be used to study the bioequivalence of a generic tablet of rifampicin 450 mg “X” marketed in Indonesia. Methods: Bioequivalence test was analysed using HPLC equipped with UV-Vis detector at 377 nm. The mobile phase used was acetonitrile-phosphate buffer pH 6.8 (45:55) delivered at a flow rate of 1.5 ml/min. Bioequivalence test was conducted on a limited number of subjects (n=8). The subjects were divided into two groups randomly. The pharmacokinetic profiles of the test tablet and reference tablet were statistically calculated using SPSS program to see the test tablet and reference tablet were bioequivalence or not. Results: The developed HPLC method for rifampicin analysis in plasma was sufficiently valid based on the International Conference on Harmonization (ICH) and European Medicines Agency (EMA) guideline, with precision and accuracy values were % Relative Standard Deviation (% RSD = 1.40–13.04) and % Recovery (86.24–102.13), respectively. Meanwhile, the method was linear over studied concentration (0.05 to 10.26 µg/ml) with a coefficient of determination (R2) = 0.9984. The method also had good stability and sensitivity. The result of statistical calculation showed that the generic rifampicin tablet X was bioequivalence toward the reference tablet Rimactan 450 mg. Conclusion: The test rifampicin tablet that was, the generic tablet “X” was bioequivalence toward the reference rifampicin tablet “Rimactan”.

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A Selective High-Performance Liquid Chromatography Method for the Determination of Reboxetine in Bulk Drug and Tablets

Journal of AOAC International ◽

10.1093/jaoac/89.6.1552 ◽

2006 ◽

Vol 89 (6) ◽

pp. 1552-1556

Author(s):

ArmaĞan Önal ◽

Olcay SaĞiri ◽

S Müge Çetin ◽

Sidika Toker

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

High Performance ◽

Depressive Disorders ◽

Degradation Products ◽

Severe Depression ◽

Hplc Method ◽

Chromatography Method ◽

Relative Standard

Abstract Reboxetine is used as a selective noradrenaline reuptake inhibitor for the treatment of major depressive disorders. It is effective in the treatment of severe depression and safer to use than traditional tricyclic antidepressants. In this study, a novel, simple, and rapid stability-indicating high-performance liquid chromatography (HPLC) method for reboxetine methansulfonate was successfully developed and validated for the assay of tablets. The method was used to quantify reboxetine in tablets; it employed a C18 column (150 4.6 mm id) with an isocratic mobile phase consisting of methanolphosphate buffer (pH 7, 0.02 M; 55 + 45, v/v) at a flow rate of 1.0 μmL/min. Reboxetine was detected by an ultraviolet detector at 277 nm. The retention time of reboxetine was about 4.5 min. The developed HPLC method was validated with respect to linearity, precision, sensitivity, accuracy, and selectivity. The method was linear over the concentration range 150 g/mL (r 0.9999). The limits of detection and the quantitation of reboxetine were 0.1 and 0.3 μg/mL, respectively. The relative standard deviation values for intraday and interday precision were 0.781.01 and 1.081.37%, respectively. Selectivity was validated by subjecting a stock solution of reboxetine to neutral, acid, and alkali hydrolysis, as well as oxidation, dry heat treatment, and photodegradation. The peaks of the degradation products did not interfere with the peak of reboxetine. The results indicated that the proposed method could be used in a stability assay. The proposed method was successfully applied to the determination of reboxetine in tablets. Excipients present in the tablets did not interfere with the analysis.

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Rapid and Sensitive Liquid Chromatographic Method for Determination of Anastrozole in Different Polymer–Lipid Hybrid Nanoparticles

SLAS TECHNOLOGY Translating Life Sciences Innovation ◽

10.1177/2472630320982308 ◽

2021 ◽

pp. 247263032098230

Author(s):

Dilshad Ahmad ◽

Faisal A. Al Meshaiti ◽

Yazeed K. Al Anazi ◽

Osama Al Owassil ◽

Alaa Eldeen B. Yassin

Keyword(s):

High Performance ◽

Limit Of Detection ◽

Reversed Phase ◽

Hplc Method ◽

Hybrid Nanoparticles ◽

Array Detector ◽

Relative Standard ◽

Validation Parameters ◽

Limit Of Quantitation ◽

Inhibitor Drug

Anastrozole, an aromatase inhibitor drug, is used for the treatment of breast cancer in pre- and postmenopausal women. Anastrozole’s incorporation into nanoparticulate carriers would enhance its therapeutic performance. To perceive the exact loaded amount of drug in nanocarriers, a valid analytical method is required. The reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated by using the C18 column, 150 × 4.6 mm, 5 µm particle size, in isocratic mobile phase composed of 50:50 V/V (volume/volume) acetonitrile–phosphate buffer (pH 3) flowing at a rate of 1.0 mL/min, and a diode array detector (DAD) set at λmax = 215 nm. The validation parameters such as linearity, accuracy, specificity, precision, and robustness have proven the accuracy of the method, with the relative standard deviation percentage (% RSD) values < 2. The limit of detection of the method was found equal to 0.0150 µg/mL, and the limit of quantitation was 0.0607 µg/mL. The percent recovery of sample was in the range of 98.04–99.25%. The method has the advantage of being rapid with a drug retention time of 2.767 min, specific in terms of resolution of peaks void of interference with any of the excipients, and high reproducibility. This makes it highly applicable for quality control purposes.

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Novel Determination of Anti-Hypertensive Combination; Benidipine Hydrochloride and Nebivolol Hydrochloride by High Performance Chromatographic Method

Research Journal of Pharmacy and Technology ◽

10.52711/0974-360x.2021.00947 ◽

2021 ◽

pp. 5433-5438

Author(s):

V.L.N. Balaji Gupta Tiruveedhi ◽

Venkateswara Rao Battula ◽

Kishore Babu Bonige ◽

Tejeswarudu B.

Keyword(s):

High Performance ◽

Degradation Products ◽

Research Work ◽

Hplc Method ◽

Assay Method ◽

Injection Quantity ◽

Relative Standard ◽

Nebivolol Hydrochloride ◽

Concentration Series

This research work was designed to establish and validate a novel stability indicating RP-HPLC method for the combined determination of Benidipine hydrochloride (BHE) and Nebivolol hydrochloride (NHE) in bulk and tablets, dependent on ICH guidelines.The assay method to analyse BHE and NHE was optimized with isocratic elution using acetonitrile: 0.1M acetate buffer (45:55, pH 5.1), Lichrospher ODS RP-18 column and flow pace of 1 ml/min. Total time for single run was 14 min. The injection quantity was 20μl, and was detected at 249nm. The method was verified on a concentration series of 1.25-10μg/ml (NHE) and 1.0-10μg/ml (BHE) for precision, accuracy and linearity. The LOD values were 0.059µg/ml and 0.028µg/ml for NHE and BHE, respectively. The LOQ values were 0.196µg/ml for NHE and 0.094µg/ml for BHE. The recovery percentages were 98.60-100.11% (BHE) and 98.94-101.50% (NHE) with relative standard deviation 0.250-0.694% (BHE) and 0.183-0.400% (NHE). The method was also observed to be efficient, and was sufficiently specific to measure BHE and NHE in the presence of stress-produced degradation products.

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Validation of high performance liquid chromatography method for determination of meloxicam loaded PEGylated nanocapsules

Brazilian Journal of Pharmaceutical Sciences ◽

10.1590/s1984-82502015000400008 ◽

2015 ◽

Vol 51 (4) ◽

pp. 823-832 ◽

Cited By ~ 1

Author(s):

Francine Rodrigues Ianiski ◽

Luciane Varini Laporta ◽

Alexandre Machado Rubim ◽

Cristiane Luchese

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

Zeta Potential ◽

Analytical Method ◽

High Performance ◽

Ph Value ◽

Hplc Method ◽

Polydispersity Index ◽

Size Distributions ◽

Relative Standard

abstract A method to ensure that an analytical method will produce reliable and interpretable information about the sample must first be validated, making sure that the results can be trusted and traced. In this study, we propose to validate an analytical high performance liquid chromatography (HPLC) method for the quantitation of meloxicam loaded PEGylated nanocapsules(M-PEGNC). We performed a validation study, evaluated parameters including specificity, linearity, quantification limit, detection limit, accuracy, precision and robustness. PEGylated nanocapsules were prepared by interfacial deposition of preformed polymer, and the particle size, polydispersity index, zeta potential, pH value and encapsulation efficiency were characterized. The proposed HPLC method provides selective, linear results in the range of 1.0-40.0 μg/mL; quantification and detection limits were 1.78 μg/mL and 0.59 μg/mL, respectively; relative standard deviation for repeatability was 1.35% and intermediate precision was 0.41% and 0.61% for analyst 1 and analyst 2, respectively; accuracy between 99.23 and 101.79%; robustness between 97.13 and 98.45% for the quantification of M-PEGNC. Mean particle diameters were 261 ± 13 nm and 249 ± 20 nm, polydispersity index was 0.15 ± 0.07 and 0.17 ± 0.06, pH values were 5.0 ± 0.2 and 5.2 ± 0.1, and zeta-potential values were -37.9 ± 3.2 mV e -31.8 ± 2.8 mV for M-PEGNC and placebo(B-PEGNC), respectively. In conclusion, the proposed analytical method is suitable for the quality control of M-PEGNC. Moreover, suspensions showed monomodal size distributions and low polydispersity index indicating high homogeneity of formulations with narrow size distributions, and appropriate pH and zeta potential. The extraction process was efficient for release of meloxicam from nanostructured systems.

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SENSITIVE DETERMINATION OF RELATED SUBSTANCES IN PIOGLITAZONE HYDROCHLORIDE BY HPLC

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2017v9i2.16828 ◽

2017 ◽

Vol 9 (2) ◽

pp. 34

Author(s):

N. Balaji ◽

Sayeeda Sultana

Keyword(s):

Mobile Phase ◽

High Performance ◽

Limit Of Detection ◽

High Performance Liquid Chromatographic ◽

Hplc Method ◽

Internal Diameter ◽

Related Substances ◽

Relative Standard ◽

Limit Of Quantitation

Objective: An efficient, high performance liquid chromatographic method has been developed and validated for the quantification of related substances in pioglitazone hydrochloride drug substance.Methods: This method includes the determination of three related substances in pioglitazone hydrochloride. The mobile phase A is 0.1% w/v triethylamine in water with pH 2.5 adjusted by dilute phosphoric acid. The mobile phase B is premixed and degassed mixtures of acetonitrile and methanol. The flow rate was 1 ml/min. The elution used was gradient mode. The HPLC column used for the analysis was symmetry C18 with a length of 250 mm, the internal diameter of 4.6 mm and particle size of 5.0 microns.Results: The developed method was found to be linear with the range of 0.006-250% with a coefficient of correlation 0.99. The precision study revealed that the percentage relative standard deviation was within the acceptable limit. The limit of detection and limit of quantitation of the impurities was less than 0.002%and 0.006% with respect to pioglitazone hydrochloride test concentration of 2000 µg/ml respectively. This method has been validated as per ICH guidelines Q2 (R1).Conclusion: A reliable, economical HPLC method was magnificently established for quantitative analysis of related substances of pioglitazone hydrochloride drug substance.

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