Response Surface Optimization for Simultanous Estimation of Sofosbuvir and Velpatasvir in Human Plasma Sample

An efficient column friendly, buffer free, highly sensitive, cost effective RP-HPLC method was developed by considering the criticality of different method parameters on analytical attributes like tailing factor, resolution and retention time in preliminary risk analysis and screening designs. The Pareto analysis of screening design highlighted the need for optimization of resolution and its influencers (capacity factor and theoretical plates) for both the drugs to imbibe quality in the method. The suggested method of optimization design was developed using ZODIAC C18 ODS (250 mm × 4.6 mm, 5 μm) column in isocratic mode using mobile phase acetonitrile : methanol : water in the ratio of 60:10:30 at a flow rate of 0.8 mL/min and UV detection wavelength of 262 nm. The retention times of drugs were found to be 3.488 minutes for sofosbuvir, 5.387 minutes for velpatasvir. The linear regression analysis data for the calibration plots showed good linear relationship with r2=0.997 for sofosbuvir, r2=0.988 for velpatasvir, in the working concentration range of 1000-5000 ng/mL, 250-1250 ng/mL respectively. The AQbD devised method was applied for quantification of drugs in plasma and validated as suggested in ICH M10 guidelines.

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Challenges of HPLC Method Development and Validation for the Assay of Bemotrizinol from Complex Matrix in Cosmeceutical Preparation

International Journal of Pharmaceutical Quality Assurance ◽

10.25258/ijpqa.11.3.2 ◽

2020 ◽

Vol 11 (03) ◽

pp. 310-316

Author(s):

Kallol S Jana ◽

Beduin Mahanti

Keyword(s):

Mobile Phase ◽

High Performance ◽

Method Development ◽

Limit Of Detection ◽

Linear Regression Analysis ◽

Cost Effective ◽

Analysis Data ◽

Hplc Method ◽

Limit Of Quantification ◽

Relative Standard

A simple high performance liquid chromatography (HPLC) method was developed for the assay of bemotrizinol (Tinosorb-S) from the complex pharmaceutical cosmetics matrix. Unlike the existing methods, the proposed mobile phase used in this method is very simple and excluding buffer. The use of buffer reducing column longevity and also a time-consuming process which increases the cost of analysis. To overcome all the referred problems, the present article was developed and validated as per International Council for Harmonization (ICH) guidelines. The reverse-phase chromatography was performed on Shimadzu model no. SPD-M10A VP with LC solution software, μBondapack (3.9 × 300 mm, 10-micron particle size) column with methanol (100%) as mobile phase at a flow rate 2.5 mL per minutes and UV detection at 254 nm. The retention time of bemotrizinol was found in 17.599 minutes, and the linear regression analysis data for the calibration plots showed a good linear relationship in the concentration range 70 to 130 μg/mL. The value of the correlation coefficient, slope, and intercept were 0.996, 7,715, and 15,320, respectively. The limit of quantification (LoQ) and limit of detection (LoD) were found to be 1.32 and 0.44, respectively. The relative standard deviation (RSD) for intra-day sample A 1.0858, sample B 0.8859, and inter-day sample A 0.9921, sample B 0.967 which were found to be lesser than 2%. The developed method was validated with regard to linearity, accuracy, precision, selectivity, and robustness, and the method was found to be simple, cost-effective, precise, accurate, linear, and specific for the successful identification and determination of bemotrizinol in pharmaceutical cosmetic preparation.

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Development and Validation of Stability-indicating RP-HPLC Method for Coumarin Assay in Bulk Drugs and Pharmaceutical Products

International Journal of Pharmaceutical Sciences and Nanotechnology ◽

10.37285/ijpsn.2013.6.3.11 ◽

2013 ◽

Vol 6 (3) ◽

pp. 2187-2192

Author(s):

S. Lakshmana Prabu ◽

S Thiyagarajan ◽

P Balan ◽

T N K Suriyaprakash ◽

Sharavanan S P

Keyword(s):

Linear Regression Analysis ◽

Analysis Data ◽

Pharmaceutical Products ◽

Dosage Forms ◽

Hplc Method ◽

Pharmaceutical Dosage Forms ◽

Stability Indicating ◽

Rp Hplc ◽

Development And Validation ◽

Simple Economic

A simple, economic, selective, precise, rapid and reproducible stability indicating HPLC method for the analysis of coumarin (CMN) in bulk drugs and from pharmaceutical formulation has been developed and validated. Separation was achieved on a symmetry C18 column using a mobile phase consisting of a mixture of methanol:water in the ratio of (70:30%v/v) at a flow rate of 1ml/min with detection at 276 nm. Linear regression analysis data for the calibration revealed a good linear relationship between response and concentration in the range 2 μgml-1 to 14 μg ml1 of CMN; the correlation coefficient was 0.999. The LOD and LOQ were 30 and 100 ng ml-1 respectively. The method was validated for accuracy, precision, recovery, reproducibility, specificity, robustness and degradation studies. The statistical analysis showed the method to be precise, reproducible, selective, specific and accurate for analysis and estimation of coumarin in bulk drugs and pharmaceutical dosage forms.

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Cefpodoxime Proxetil: A New Stability Indicating RP-HPLC Method

ISRN Chromatography ◽

10.1155/2013/328157 ◽

2013 ◽

Vol 2013 ◽

pp. 1-8 ◽

Cited By ~ 4

Author(s):

Ceema Mathew ◽

M. Ajitha ◽

P. R. Sathesh Babu

Keyword(s):

High Performance ◽

Uv Light ◽

Degradation Products ◽

Linear Regression Analysis ◽

Analysis Data ◽

Hplc Method ◽

Uv Detector ◽

Standard Drug ◽

Cefpodoxime Proxetil ◽

Stability Indicating

The present work describes the development of a sensitive and economic stability indicating high performance liquid chromatographic (HPLC) method for the determination of cefpodoxime proxetil (CP) as bulk drug and as pharmaceutical formulation. Both R and S isomers of the drug were separated using Phenomenex ( mm, 5 μm particle size) ODS column with a flow rate of 1 mL min−1 and an SPD 20 A UV detector to monitor the eluate at 252 nm. The isocratic method used a mobile phase consisting of methanol and phosphate buffer of pH 4.0 in the ratio 65 : 35. The linear regression analysis data for the calibration plots showed good linear relationship with in the working concentration range of 5–100 μg mL−1. The LOD and LOQ were 53 and 160 ng mL−1, respectively. CP was subjected to stress degradation using acid, alkali, hydrogen peroxide, dry heat, wet heat, and UV light. The standard drug peaks were well resolved from the degradation products’ peaks with significantly different retention time (Rt), and the resolution factor for the R and S isomers of CP was found to be greater than 2.

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RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATION OF DESVENLAFAXINE SUCCINATE BULK DRUG IN ARTIFICIAL URINE

INDIAN DRUGS ◽

10.53879/id.54.05.10728 ◽

2017 ◽

Vol 54 (05) ◽

pp. 37-40

Author(s):

A. S. Bansode ◽

◽

V. D. Shelke ◽

S. S. Gaikwad

Keyword(s):

Method Development ◽

Depressive Disorders ◽

Solid Phase ◽

Linear Regression Analysis ◽

Analysis Data ◽

Hplc Method ◽

Relative Standard ◽

Bulk Drug ◽

Hplc Method Development ◽

Development And Validation

Desvenlafaxine (DSV) succinate is a novel serotonin (5HT) and nor-epinephrine reuptake inhibitor (SNRI), which is currently used for the treatment of major depressive disorders and is being studied for use in the management of vasomotor symptoms in postmenopausal women. DSV is a major active metabolite of venlafaxine. DSV has only 30 % of protein binding and approximately 45% of the total oral dose of DSV is excreted unchanged in the urine. The chromatographic separation was performed with acetonitrile and phosphate buffer in the ratio of 25:75 (v/v) at a flow rate of 1 ml/min with UV detection at 224 nm. The extraction was done using C8 solid phase cartridges. The method is validated for precision, linearity, recovery and stability as per the USFDA guideline and the results met the acceptance criteria. The linear regression analysis data for the calibration plots showed a good linear relationship (R2 = 0.995) over a concentration range of 5-30 ppm. The percentage relative standard deviation (% RSD) values of precision were

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Application of an HPLC Method for Selective Determination of Phenazopyridine Hydrochloride: Theoretical and Practical Investigations

Journal of AOAC International ◽

10.5740/jaoacint.16-0301 ◽

2017 ◽

Vol 100 (5) ◽

pp. 1400-1406 ◽

Cited By ~ 1

Author(s):

Khalid A M Attia ◽

Nasr M El-Abasawi ◽

Ahmed El-Olemy ◽

Ahmed H Abdelazim

Keyword(s):

Density Functional ◽

Linear Regression Analysis ◽

Analysis Data ◽

Practical Investigations ◽

Reversed Phase ◽

Hplc Method ◽

Calibration Plot ◽

Good Resolution ◽

Selective Determination

Abstract HPLC method was developed for the selective determination of phenazopyridine hydrochloride (PAP) in the presence of its computationally selected metabolite. Density functional theory was applied as a computational model to study the energy of PAP metabolites, and the results revealed that 2,3,6-triaminopyridine (TAP) is the most stable metabolite. Good resolution and separation of PAP from TAP was achieved using a reversed-phase BDS Hypersil C18 column with a mobile phase consisting of acetonitrile–water (75 + 25, v/v) at flow rate of 1 mL/min and with UV detection at 280 nm. The linear regression analysis data for the calibration plot of PAP showed a good linear relationship over the concentrationrange of 5–45 μg/mL, with an LOD of 0.773 μg/mL. Moreover, a theoretical investigation of the relationship between the stationary phaseand the studied molecules was performed to confirm the experimental results. The proposed method was successfully applied for the selective determination ofPAP in pharmaceutical formulation. In addition, the obtained results were statistically compared to a reported method, with no significant differences foundbetween the investigated method and the reported onewith respect to accuracy and precision.

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Estimation of Catechin in Ayurvedic Oil Formulations Containing Acacia catechu

Journal of AOAC International ◽

10.1093/jaoac/92.4.1021 ◽

2009 ◽

Vol 92 (4) ◽

pp. 1021-1026 ◽

Cited By ~ 3

Author(s):

Nidhi Dubey ◽

Nitin Dubey ◽

Rajendra Mehta ◽

Ajay Saluja

Keyword(s):

Formic Acid ◽

Limit Of Detection ◽

Linear Regression Analysis ◽

Cost Effective ◽

Analysis Data ◽

Calibration Plot ◽

Tlc Plates ◽

Acacia Catechu ◽

Hptlc Method

Abstract A sensitive, simple, rapid, and efficient HPTLC method was developed and validated for the analysis of catechin in marketed Ayurvedic oil formulations containing Acacia catechu. Chromatography of methanolic0.1 formic acid (7:3, v/v) extracts of these formulations was performed on silica gel 60 F254 aluminum-backed TLC plates of 0.2 mm layer thickness. The plate was developed up to 85 mm with the ternarymobile phase chloroformacetone0.1 formic acid (7.7 + 1.5 + 0.8, v/v/v) at 22 2C with 20 min of chamber saturation. The system produced compact spots of catechin at an Rf value of 0.36. The marker, catechin, was quantified at its maximum absorbance of 296 nm. The limit of detection and quantitation values were 6 and 20 ng/spot, respectively. The linear regression analysis data for the calibration plot showed a good linear relationship with a correlation coefficient of 0.9993 in the concentration range of 2001200 ng/spot for catechin with respect to peak area. Repeatability of the method was 0.88 RSD. Recovery values from 97 to 102 indicate excellent accuracy of the method. The developed HPTLC method is accurate, precise, and cost-effective, and it can be successfully applied for the determination of catechin in marketed Ayurvedic oil formulations containing Acacia catechu.

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Development and Validation of Stability-Indicating HPLC Method for the Simultaneous Determination of Paracetamol, Tizanidine, and Diclofenac in Pharmaceuticals and Human Serum

Journal of AOAC International ◽

10.1093/jaoac/94.1.150 ◽

2011 ◽

Vol 94 (1) ◽

pp. 150-158 ◽

Cited By ~ 11

Author(s):

Farhan Ahmed Siddiqui ◽

M Saeed Arayne ◽

Najma Sultana ◽

Faiza Qureshi

Keyword(s):

Human Serum ◽

Simultaneous Determination ◽

Ammonium Carbonate ◽

Linear Regression Analysis ◽

Pharmaceutical Formulations ◽

Analysis Data ◽

Hplc Method ◽

Hplc Separation ◽

Development And Validation

Abstract A method is described for the simultaneous determination of paracetamol, tizanidine, and diclofenac in mixtures. The method was based on HPLC separation of the three drugs followed by UV detection at 254 nm. The separation was carried out on a Hypersil ODS, C18 (250 × 4.6 mm id, 10 μm particle size) column using the mobile phase aqueous 0.2% ammonium carbonate–methanol (60 + 40, v/v) at a flow rate of 1 mL/min. The linear regression analysis data were used for the regression curve in the range of 170–10,000 ng/mL for paracetamol, 120–10,000 ng/mL for tizanidine, and 20–10,000 ng/mL for diclofenac. No chromatographic interference from tablet excipients was found. In order to check the selectivity of the proposed method, degradation studies were carried out using hydrolysis (acid, basic, and neutral), thermolysis, and oxidation. The developed method, after being validated in terms of precision, robustness, recovery, LOD, and LOQ, was successively applied to the analysis of pharmaceutical formulations and human serum.

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Optimization and Validation of a Fast UPLC Method for Simultaneous Determination of Hydroquinone, Kojic Acid, Octinoxate, Avobenzone, BHA, and BHT

Journal of AOAC International ◽

10.5740/jaoacint.16-0104 ◽

2017 ◽

Vol 100 (1) ◽

pp. 1-7 ◽

Cited By ~ 5

Author(s):

Fanny Galimany-Rovira ◽

Pilar Pérez-Lozano ◽

Encarna García-Montoya ◽

Montse Miñarro-Carmona ◽

Josep R Ticó-Grau ◽

...

Keyword(s):

Simultaneous Determination ◽

Kojic Acid ◽

Linear Regression Analysis ◽

Extraction Procedure ◽

Analysis Data ◽

Hplc Method ◽

Butylated Hydroxytoluene ◽

Array Detector ◽

Skin Whitening

Abstract A previously published HPLC method for the simultaneous determination of six major components (hydroquinone, kojic acid, octinoxate, avobenzone, butylated hydroxyanisole, and butylated hydroxytoluene) in a skin-whitening cream was transferred and optimized to an ultra-performance LC system. Separation was achieved in a ZORBAX SB-Phenyl Rapid-Resolution High Throughput column (2.1 × 100 mm, 1.8 μm), using a mobile phase consisting of water with 0.1% acetic acid and acetonitrile at a flow rate of 0.7 mL/min. The column was maintained at 40°C, and detection was carried out at 230 nm using a diode-array detector. These chromatographic conditions allow the separation of the six compounds in 3 min instead of 14 min. The extraction procedure was optimized, reducing the time and demonstrating its suitability. The method was validated according to International Conference on Harmonization guidelines, with respect to specificity, precision, accuracy, and linearity. Selectivity was found to be satisfactory. Linear regression analysis data for all compounds showed a good linear relationship, with r2 > 0.999 in the concentration range of 50–120% of the label claim for each compound. The RSD for precision and accuracy of the method was found to be less than 2% forall compounds. Comparison of system performance with the previously published HPLC method was made with respect to analysis time, efficacy, and resolution. The proposed method is faster and consumes less solvent and was applied in the determination of six major compounds in batches of skin-whitening cream manufactured during thevalidation process.

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DEVELOPMENT AND VALIDATION OF A STABILITY INDICATING HPLC METHOD FOR THE DETERMINATION OF NILOTINIB HYDROCHLORIDE IN BULK AND PHARMACEUTICAL DOSAGE FORM

International Journal of Pharmacy and Pharmaceutical Sciences ◽

10.22159/ijpps.2016v8i9.11637 ◽

2016 ◽

Vol 8 (9) ◽

pp. 41

Author(s):

Ramu Ivaturi ◽

T. Manikya Sastry ◽

S. Satyaveni

Keyword(s):

Linear Regression Analysis ◽

Analysis Data ◽

Dosage Forms ◽

Hplc Method ◽

Pharmaceutical Dosage Form ◽

Pharmaceutical Dosage Forms ◽

Stability Indicating ◽

Rp Hplc ◽

Ich Guidelines

Objective: To develop a rapid, accurate, linear, sensitive and stability indicating RP-HPLC method for the determination of nilotinib in bulk and pharmaceutical dosage forms in the presence of its four related substances.Methods: The RP-HPLC method was developed for the chromatographic separation of nilotinib and its impurities by using waters Xterra RP-18 (150*4.6 mm, 3.5 µm) column with a mobile phase combination of 10 mM ammonium formate with pH-3.5 and acetonitrile in gradient mode. An injection volume of 20 µl. Flow rate was 1.0 ml/min and detection was carried a wavelength of 250 nm. The method was validated as per ICH guidelines.Results: The retention time for nilotinib and its four impurities were found to be 4.37, 7.40, 8.96, 10.21 and 10.87 min respectively. The linear regression analysis data for the calibration plots showed the good linear relationship in the concentration range of 0.04-3.0 ppm for the nilotinib impurities. The % recovery of nilotinib impurities was found to be 96.8-99.4% in the linearity range. The detection limit (LOD) values were about 0.014, 0.016, 0.005 and 0.03 ppm respectively and the quantification limit (LOQ) values were 0.042, 0.048, 0.014 and 0.09 ppm respectively. The % degradation at various stress conditions like acid, alkaline, oxidative, thermal and photolytic stress was found to be 8.92, 18.35,5.63, 0.88 and 3.89 respectively.Conclusion: The RP-HPLC method compatible with LC-MS was developed for the analysis of nilotinib and its four impurities. It was validated as per the ICH guidelines and found to be linear, robust, precise, accurate, sensitive, stability indicating and can be used for routine as well as stability analysis of capsule dosage forms as well as for drug substance.

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Estimation of Berberine in Ayurvedic Formulations Containing Berberis aristata

Journal of AOAC International ◽

10.1093/jaoac/91.5.1149 ◽

2008 ◽

Vol 91 (5) ◽

pp. 1149-1153 ◽

Cited By ~ 7

Author(s):

Kedar Kumar Rout ◽

Subhalaxmi Pradhan ◽

Sagar Kumar Mishra

Keyword(s):

High Performance ◽

Limit Of Detection ◽

Linear Regression Analysis ◽

Cost Effective ◽

Analysis Data ◽

Calibration Plot ◽

Methanolic Extracts ◽

Limit Of Quantitation ◽

Tlc Plates ◽

Hptlc Method

Abstract A sensitive, simple, rapid, and efficient high-performance thin-layer chromatographic (HPTLC) method has been developed and validated for the analysis of berberine in marketed Ayurvedic formulations containing Berberis aristata DC for regulatory purposes. Chromatography of methanolic extracts of these formulations was performed on silica gel 60 F254 aluminum-backed TLC plates of 0.2 mm layer thickness. The plate was developed up to 66 mm with the ternary-mobile phase butanolacetic acidwater (8 + 1 + 1, v/v/v) at 33 5C with 5 min of tank saturation. The marker, berberine, was quantified at its maximum absorbance of 350 nm. The limit of detection and limit of quantitation values were found to be 5 and 10 ng/spot. The linear regression analysis data for the calibration plot showed a good linear relationship with correlation coefficient 0.9994 in the concentration range of 10 to 50 ng/spot for berberine with respect to peak area. The instrumental precision was found to be 0.49 coefficient of variation (CV), and repeatability of the method was 0.73 CV. Recovery values from 98.27 to 99.11 indicate excellent accuracy of the method. The developed HPTLC method is very accurate, precise, and cost-effective, and it has been successfully applied to the assay of marketed formulations containing B. aristata for determination of berberine.

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