Robust optimization of gradient RP HPLC method for simultaneous determination of ivabradine and its eleven related substances by AQbD approach

AbstractThis paper is aimed at developing a gradient elution reversed-phase high-performance liquid chromatography (RP-HPLC) method for the separation of a complex mixture composed of ivabradine and its eleven impurities, in a reasonable timeframe. In order to obtain a robust and reliable HPLC method for separation of this mixture, Analytical Quality by Design (AQbD) was applied. This approach demonstrated to be useful in development of a long lasting life cycle methods. Four chromatographic variables were defined as key method parameters (KMPs) and optimized towards the analytical target profile (ATP). Designated KMPs were initial and final amount of acetonitrile in the mobile phase, pH value of the aqueous phase and gradient time, while resolutions of critical peak pairs were denoted as critical method attributes (CMAs). Relationships between KMPs and CMAs were obtained with the aid of Design of Experiments (DoEs) methodology among which Box-Behnken design (BBD) was employed to gain valid mathematical models. Obtained mathematical equations were used to construct the Design Space (DS) and select reliable optimal separation conditions. They included 11% (v/v) and 34% (v/v) of initial and final amount of acetonitrile, respectively, as well as 45 min of gradient elution time and 20 mM ammonium acetate as aqueous mobile phase with pH set to 7.35. The possibility to separate the diastereoisomers of impurity X was also evaluated. It was demonstrated that this separation could not be achieved in gradient elution mode within the defined variable domains and in a reasonable time span. The developed method was validated according to ICH Q2 (R1) guideline and met all the required criteria.

Download Full-text

A simple isocratic RP-HPLC method for quality control of oseltamivir capsules

Macedonian Journal of Chemistry and Chemical Engineering ◽

10.20450/mjcce.2012.10 ◽

2012 ◽

Vol 31 (2) ◽

pp. 205

Author(s):

Agim Ameti ◽

Jasmina Slavkovska ◽

Katerina Starkoska ◽

Zorica Arsova-Sarafinovska

Keyword(s):

Mobile Phase ◽

Limit Of Detection ◽

Degradation Products ◽

Reversed Phase ◽

Hplc Method ◽

Sample Treatment ◽

Limit Of Quantification ◽

Elution Time ◽

Rp Hplc

A simple isocratic reversed-phase high performance liquid chromatographic (RP-HPLC) method was developed for determination of oseltamivir active pharmaceutical ingredient (API) in bulk drug and pharmaceuticals. The separation was achieved on a Purospher STAR® RP – 18e column with a mobile phase consisting of methanol- 0.02 mol l-1 phosphate buffer, pH 5, 50:50 (v/v). Chromatographic results demonstrated the specificity of the method for determination of oseltamivir in presence of degradation products generated in studies of forced decomposition. The limit of detection (LOD) and limit of quantification (LOQ) for oseltamivir phosphate were 0,0162 μg ml-1 and 0,0491 μg ml-1, respectively. The advantages of this method include simple sample treatment and short elution time (less than 6 min). Furthermore, using methanol instead of acetonitrile in a mobile phase composition considerably reduces the laboratory expenses, still retaining adequate sensitivity for routine analysis as well as for evaluation of potentially counterfeit Tamiflu® products.

Download Full-text

SIMULTANEOUS ESTIMATION OF CURCUMIN AND GEFITINIB IN BULK AND TISSUE SAMPLES (PLASMA AND BRAIN HOMOGENATE) BY RP-HPLC: APPLICATION TO A DISTRIBUTION STUDY

INDIAN DRUGS ◽

10.53879/id.56.07.11330 ◽

2019 ◽

Vol 56 (07) ◽

pp. 59-68

Author(s):

H Mahajan ◽

S Savale ◽

P Nerkar ◽

Keyword(s):

Flow Rate ◽

Mobile Phase ◽

Internal Standard ◽

Brain Homogenate ◽

Reversed Phase ◽

Hplc Method ◽

Simultaneous Estimation ◽

Bulk Analysis ◽

Experimental Conditions ◽

Rp Hplc

The present study was aimed at developing a Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) method for simultaneous determination of curcumin (CRM) and gefitinib (GFT) in bulk, plasma and brain homogenate. hydrochlorothiazide was used as an internal standard (IS). A new simple, rapid, selective, precise and accurate RP-HPLC method has been developed. The separation was achieved by using C-18 column (Qualisil BDS C18, 250 mm x 4.6 mm I.D.) coupled with a guard column of silica, mobile phase consisted of acetonitrile: water with 0.1% formic acid (30:70 v/v). The flow rate was 0.2 ml/min and the drug was detected using PDA detector at the wavelength of 242 nm. The experimental conditions, including the diluting solvent, mobile phase composition, column saturation and flow rate, were optimised to provide high-resolution and reproducible peaks. The method was developed and tested for linearity range of 10-60 μg/mL for bulk analysis and 200-800 ng/mL for plasma and brain homogenate. The developed method was validated as per ICH guidelines, in terms of linearity, application of the proposed method to bulk sample, recovery, precision, repeatability, ruggedness, sensitivity (LOD and LOQ) and robustness and stability study (short and long-term stabilities, freeze/thaw stability, post-preparative). The low value of % RSD showed that the method was precise within the acceptance limit of 2%. The developed method was successfully applied for the analysis of the drug in bulk as well as various marketed formulation and drug in plasma and brain distribution studies.

Download Full-text

Application of a reversed-phase HPLC method for quantitative p-coumaric acid analysis in wine

OENO One ◽

10.20870/oeno-one.2010.44.2.1465 ◽

2010 ◽

Vol 44 (2) ◽

pp. 117

Author(s):

Dominique Salameh ◽

Cédric Brandam ◽

Toufic Rizk ◽

Roger Lteif ◽

Pierre Strehaiano

Keyword(s):

Mobile Phase ◽

Chromatographic Method ◽

Red Wine ◽

Reversed Phase ◽

Hplc Method ◽

Coumaric Acid ◽

Reversed Phase Hplc ◽

Mobile Phase Composition ◽

Rp Hplc ◽

Synthetic Media

Aims: This paper presents a rapid chromatographic method to monitor the concentration of p-coumaric acid in wine and in bioconversion studies.Methods and results: RP-HPLC method was validated in synthetic wine medium and in natural red wine. Mobile phase composition was water 77%, acetonitrile 23%. Formic acid was added to control pH at 3.5. The flow was 0.7 mL/min and the temperature 30 °C. The detection was done using UV at 305 nm. The linearity range was validated between 0.5 and 15 mg/L. The resolution was respectively 5.35 and 2.99. The detection and quantification limits were 0.01 mg/L and 0.04 mg/L. This method was used to study p-coumaric acid bioconversion into 4-ethylphenol and 4-vinylphenol, and to study this acid adsorption in enological conditions.Conclusions: This paper presented a simple HPLC method to monitor the concentration of p-coumaric acid in synthetic media and natural wine. It was used to study the p-coumaric acid bioconversion rates and mechanism.Significance and impact of the study: This method is useful to monitor p-coumaric acid concentration, which helps to predict amounts of 4-ethylphenol or 4-vinylphenol that can be produced in wine. This method can be helpful to control undesirable phenolic flavors potential in wine.

Download Full-text

Development of RP-HPLC Method for the Quantitative Estimation of Ofloxacin and Ornidazole in Combined Liquid Oral Dosage Forms

International Journal of Pharmaceutical Sciences and Nanotechnology ◽

10.37285/ijpsn.2013.6.1.8 ◽

2013 ◽

Vol 6 (1) ◽

pp. 1972-1976

Author(s):

Cylma Menezes ◽

Varun G ◽

Shyamkumar B ◽

Reema N

Keyword(s):

Mobile Phase ◽

Dynamic Range ◽

Quantitative Estimation ◽

Reversed Phase ◽

Dosage Forms ◽

Hplc Method ◽

Oral Dosage ◽

Rp Hplc ◽

Validation Parameters ◽

Oral Dosage Forms

A simple, efficient and reproducible reversed phase high performance liquid chromatographic (RP-HPLC) method for the quantitative estimation of ofloxacin and ornidazole in bulk and in combined liquid oral dosage form has been developed and validated. The separation was carried out using thermohypersil phenyl column (250 mm × 4.6 mm, 5 μm) as stationary phase with isocratic flow and phosphate buffer (adjusted to pH 2.4 with ortho phosphoric acid): acetonitrile (87:13 v/v) as mobile phase. Mobile phase was maintained at a flow rate of 1.0 ml/min and UV detection was carried out at 294 nm. The retention time of ofloxacin and ornidazole was 10.40 and 5.69 min, respectively. All calibration curves showed good linear correlation coefficients within the tested limits (r2 > 0.9995). The linear dynamic range was 10-100 µg/ml and 25-250 µg/ml for ofloxacin and ornidazole respectively. Percentage recoveries for ofloxacin and Ornidazole were 100.48 % and 99.84 % respectively. All the analytical validation parameters were determined and found in the limit as per the International Conference on Harmonization (ICH) guidelines, which indicates the validity of the method. The validated method is also found to be accurate, precise and robust for the quantitative estimation of ofloxacin and ornidazole in combined liquid oral dosage forms.

Download Full-text

Development and Validation of a Reversed-Phase HPLC Method for Determination of Alkaloids from Papaver somniferum L. (Papaveraceae)

Journal of AOAC International ◽

10.5740/jaoacint.11-102 ◽

2012 ◽

Vol 95 (2) ◽

pp. 399-405 ◽

Cited By ~ 12

Author(s):

Jelena Acevska ◽

Aneta Dimitrovska ◽

Gjoshe Stefkov ◽

Katerina Brezovska ◽

Marija Karapandzova ◽

...

Keyword(s):

Mobile Phase ◽

Ph Value ◽

Reversed Phase ◽

Hplc Method ◽

Papaver Somniferum ◽

Chromatographic Behavior ◽

Organic Modifier ◽

Column Temperature ◽

Development And Validation

Abstract An HPLC method for the separation of six target alkaloids from Papaver somniferum L. (morphine, codeine, oripavine, thebaine, papaverine, and noscapine) was developed, optimized, and validated. The chromatographic behavior of these alkaloids was investigated using a reversed-phase chromatography at acidic and alkaline pH. The effects of ion-pairing agents, pH value of the mobile phase, concentration of the buffer components, mobile phase organic modifier, and column temperature were studied. Regardless of the large differences in their pKa values, all alkaloids were separated within a close retention window, and good peak shape was achieved for each of the six alkaloids. The proposed method has adequate selectivity, linearity, accuracy, precision, and reproducibility and is applicable for poppy straw.

Download Full-text

Optimization of RP-HPLC method with UV detection for determination of ursodeoxycholic acid in pharmaceutical formulations

Macedonian Pharmaceutical Bulletin ◽

10.33320/maced.pharm.bull.2020.66.01.010 ◽

2020 ◽

Vol 66 (1) ◽

pp. 85-90

Author(s):

Zhaklina Poposka Svirkova ◽

Zorica Arsova-Sarafinovska ◽

Aleksandra Grozdanova

Keyword(s):

Ursodeoxycholic Acid ◽

Flow Rate ◽

Mobile Phase ◽

Pharmaceutical Formulations ◽

Gradient Elution ◽

Hplc Method ◽

Uv Detection ◽

Rp Hplc ◽

Ich Guidelines

Due to the low absorptivity of bile acids, the aim of this study was to develop and validate a simple and sensitive HPLC/UV method for quantification of ursodeoxycholic acid (UDCA) in pharmaceutical formulations. Effective separation was achieved on C18 end–capped column, with gradient elution of a mobile phase composed of 0.001 M phosphate buffer (pH 2.8±0.5) – acetonitrile mix, at flow rate 1.5 mL min-1, UV detection at 200 nm and injection volumes were 50 µL. The proposed HPLC method was fully validated according to the ICH guidelines and it was found to be simple, accurate, precise and robust. Key words: ursodeoxycholic acid, HPLC/UV, pharmaceutical formulations, validation

Download Full-text

ANALYTICAL QUALITY-BY-DESIGN (AQBD) APPROACH FOR THE DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR THE ESTIMATION OF LAMOTRIGINE IN BULK AND TABLET FORMULATION

Journal of medical pharmaceutical and allied sciences ◽

10.22270/jmpas.v10i5.1506 ◽

2021 ◽

Vol 10 (5) ◽

pp. 3591-3596

Author(s):

Manisha P. Puranik

Keyword(s):

Flow Rate ◽

Mobile Phase ◽

High Performance ◽

Quality By Design ◽

Active Pharmaceutical Ingredient ◽

Reversed Phase ◽

Hplc Method ◽

Analytical Technique ◽

Rp Hplc ◽

Ich Guideline

The current analytical exploration illustrated developing a reversed-phase high-performance liquid chromatography (RP-HPLC) technique and consequent substantiation for analyzing lamotrigine (LAM) active pharmaceutical ingredient (API) using a Quality-by-design (QbD) approach (Central Composite Design), in bulk product as well as in the tablet formulations. In this experiment, based on systematic scouting, four key components (viz., mobile phase, column, flow-rate, and wavelength) were studied by the RP-HPLC method. 13 experimental runs were done with acetonitrile (ACN) (40-60% v/v) having flow-rate in the range 0.8 mL/min to 1.2 mL/min. The proposed analytical method was thoroughly corroborated in terms of ruggedness linearity, robustness, accuracy, and precision in accordance with ICH guideline Q2A and ICH guideline Q2B. Under the optimum chromatographic environment; Intersil C8 column of 250 mm length, 4.6 mm (i.d.); 20 μL injection volume; and mobile phase ACN: Methanol (60:40 v/v), a retention time of 2.542 min was noticed at 220 nm detection wavelength. The method was found to be extremely reproducible, accurate, linear, precise, robust, and economically adequate to execute the estimation. The intended analytical technique was thoroughly assessed through statistical tools and could be an imperative concern for the habitual scrutiny of LAM in bulk products and its formulation.

Download Full-text

Development and Validation of RP-HPLC Method for Desoximetasone in Bulk and Cream Formulation

Journal of Drug Delivery and Therapeutics ◽

10.22270/jddt.v9i3.2630 ◽

2019 ◽

Vol 9 (3) ◽

pp. 154-159

Author(s):

Ashish Gorle ◽

Jayashri Mahajan ◽

Prathamesh Bhave

Keyword(s):

Method Validation ◽

Mobile Phase ◽

High Performance ◽

Gradient Elution ◽

Hplc Method ◽

Topical Steroids ◽

Cream Formulation ◽

Rp Hplc ◽

Validation Parameters ◽

Development And Validation

Desoximetasone chemically is 9-fluoro-IIβ21-dihydroxy-I6a-methylpregna-I.4-diene-3.The precise mechanism of the anti-inflammatory activity of topical steroids in the treatment of steroid-responsive dermatoses, in general, is uncertain. So, in present investigation chromatographic methods were developing use RP-HPLC for estimation of Desoximetasone in bulk and in cream formulation and method validation according to ICH guidelines. The main objective of this study was to develop a simple and reproducible method for desoximetasone by Reverse Phase High Performance Liquid Chromatography (RP-HPLC). In this work the desoximetasone separation was carried out by using C18 cosmosile column (250mmx4.6mm particle size 5µm). By using 0.1% orthrophosphoric acid pH adjusted up to 3 at uv detection of 240nm.The mobile phase was used at various ratio for gradient elution the ratio of mobile phase was 20:80 v/v. Methanol and water used for mobile phase and flow rate was being set at 1mL/min. The linearity of proposed method was found in range of r =0.9989. Statistically validation parameters such as linearity, accuracy, precision, LOD and LOQ were checked. Keywords: Desoximetasone, RP-HPLC, Method validation.

Download Full-text

DEVELOPMENT AND VALIDATION OF REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR GABAPENTIN AND ITS RELATED SUBSTANCES IN CAPSULE DOSAGE FORM AND EXCIPIENT COMPATIBILITY STUDIES

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2018.v11s4.31707 ◽

2018 ◽

Vol 11 ◽

Author(s):

Afroz Patan

Keyword(s):

Mobile Phase ◽

High Performance ◽

Dosage Form ◽

Reversed Phase ◽

Hplc Method ◽

Compatibility Studies ◽

Compound A ◽

Related Substances ◽

Rp Hplc ◽

Excipient Compatibility

Objective: A simple, accurate, precise, and reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for gabapentin (GBP) and its related substances in the capsule dosage form and excipient compatibility studies. Methods: The review of literature indicates that various methods have been reported for the estimation of GBP. When some excipients were used for GBP, it produced degradation product called lactam due to the presence of more water content. Hence, a novel RP-HPLC method has been developed for studying excipient compatibility and related substances of GBP in capsule dosage form using excipients such as lactose anhydrous and dried maize starch which is having less water activity. Waters Alliance e2695 separation module with ultraviolet/photodiode array (UV/PDA) detector with Inertsil C8 (250 mm×4.6 mm); 5 μm with an injection volume of 50 μl is injected and eluted with the (gradient program) mobile Phase A buffer: acetonitrile (940:60) and mobile phase B buffer: acetonitrile (700:300) pH 6.9 with 5 N potassium hydroxide which is pumped at a speed of 1.5 ml/min and detected by UV/PDA detector at 210 nm. The peaks of GBP and GBP-related compound A are well separated at 6.7 min and 34.5 min, respectively. Results: The method developed was approved for various parameters such as accuracy, specificity, precision, intermediate precision, range, linearity, robustness, limit of detection, limit of quantification, steadiness, and system suitability according to the International Conference on Harmonization guidelines. The results got were according to the acceptance criteria. Conclusion: The technique proposed was assured for detection of related substances in the marketed formulation and could be used for the routine analysis of GBP and GBP-related compound A in the capsule dosage form.

Download Full-text

Standardization of Adhatoda vasica Nees Market Preparations by RP-HPLC Method

Dhaka University Journal of Pharmaceutical Sciences ◽

10.3329/dujps.v15i1.29193 ◽

2016 ◽

Vol 15 (1) ◽

pp. 57-62 ◽

Cited By ~ 1

Author(s):

BK Sajeeb ◽

Uttom Kumar ◽

Md Shahadat Hossain ◽

Sitesh C Bachar

Keyword(s):

Mobile Phase ◽

Reversed Phase ◽

Hplc Method ◽

Reversed Phase Hplc ◽

Reference Standard ◽

Adhatoda Vasica ◽

Traditional Medicines ◽

Marker Compound ◽

Rp Hplc ◽

Qualitative Analyses

In recent times, quality control of herbal and traditional medicines with modern scientific techniques and knowledge are of great concern. The present study reveals a simple and improved reversed phase HPLC method for qualitative analyses of Adhatoda vasica Nees market preparations via quantitation of its major metabolite, vasicine as a marker compound. Three market preparations, each of four different herbal and traditional manufacturers, were analyzed. The market preparations were extracted with chloroform and the residue obtained from extraction of each market preparation was analyzed for quantitation of vasicine by RP-HPLC method with ODS column using a mixture of water and methanol (60:40) as mobile phase at a flow rate of 0.5 ml/min. The estimated quantities of vasicine compared to reference standard for marketed products of four different manufacturers were found to be 1.502 ± 0.064 g, 1.590 ± 0.081 g, 1.761 ± 0.061 g and 1.627 ± 0.082 g, respectively per 100 ml of preparation.Dhaka Univ. J. Pharm. Sci. 15(1): 57-62, 2016 (June)

Download Full-text