Full optimization and validation of an HPLC method for the quantitative analysis of total sugars in a soft drink

2020 ◽  
Vol 34 (2) ◽  
pp. 419-426
Author(s):  
V. Armoogum ◽  
K. Boodhoo
Keyword(s):  
Quantitative Analysis ◽  
Mobile Phase ◽  
Soft Drink ◽  
Hplc Method ◽  
Total Sugars ◽  
System Suitability ◽  
Accuracy Check ◽  
Full Optimization ◽  
Refractive Index Detector ◽  
Optimum Separation

Five HPLC methods were employed for the quantitative analysis of three natural sugars namely fructose, glucose and sucrose in soft drinks. HPLC-refractive index detector (RID)-AMINO proved to be the most suitable HPLC method to carry out the latter task. For the optimum separation and response of the natural sugars the best conditions employed were column oven temperature 30 oC, flow rate 0.1 mL/min, mobile phase ratio acetonitrile:water 75:25 and they were determined by studying all possible interactions among these three parameters. Full validation of HPLC-RID-AMINO was performed in terms of system suitability test, precision check, accuracy check and robustness.                     KEY WORDS: Sugar, Soft drink, Experimental design, Validation, HPLC, System suitability   Bull. Chem. Soc. Ethiop. 2020, 34(2), 419-426 DOI: https://dx.doi.org/10.4314/bcse.v34i2.17

scholarly journals Box-Behnken Assisted Validation and Optimization of an RP-HPLC Method for Simultaneous Determination of Domperidone and Lansoprazole

Separations ◽  
2021 ◽  
Vol 8 (1) ◽  
pp. 5
Author(s):  
Mohd Afzal ◽  
Mohd. Muddassir ◽  
Abdullah Alarifi ◽  
Mohammed Tahir Ansari
Keyword(s):  
Flow Rate ◽  
Mobile Phase ◽  
Limit Of Detection ◽  
Hplc Method ◽  
Box Behnken Design ◽  
Rp Hplc ◽  
System Suitability ◽  
Method Optimization ◽  
Key Variables

A highly specific, accurate, and simple RP-HPLC technique was developed for the real-time quantification of domperidone (DOMP) and lansoprazole (LANS) in commercial formulations. Chromatographic studies were performed using a Luna C8(2), 5 μm, 100Å, column (250 × 4.6 mm, Phenomenex) with a mobile phase composed of acetonitrile/2 mM ammonium acetate (51:49 v/v), pH 6.7. The flow rate was 1 mL·min−1 with UV detection at 289 nm. Linearity was observed within the range of 4–36 µg·mL−1 for domperidone and 2–18 µg·mL−1 for lansoprazole. Method optimization was achieved using Box-Behnken design software, in which three key variables were examined, namely, the flow rate (A), the composition of the mobile phase (B), and the pH (C). The retention time (Y1 and Y3) and the peak area (Y2 and Y4) were taken as the response parameters. We observed that slight alterations in the mobile phase and the flow rate influenced the outcome, whereas the pH exerted no effect. Method validation featured various ICH parameters including linearity, limit of detection (LOD), accuracy, precision, ruggedness, robustness, stability, and system suitability. This method is potentially useful for the analysis of commercial formulations and laboratory preparations.

scholarly journals HPLC Quantitative Analysis of Main Stilbenes and Flavones in Different Parts ofMatteuccia struthiopteris

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Shubei Li ◽  
Dong Zhang ◽  
Lan Yang ◽  
Yujie Li ◽  
Xiaoxin Zhu ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Linear Regression ◽  
Phosphoric Acid ◽  
Flow Rate ◽  
Mobile Phase ◽  
Gradient Elution ◽  
Hplc Method ◽  
Regression Coefficients ◽  
The Mean ◽  
Different Parts

A simple and accurate HPLC-UV method was developed for the simultaneous quantitative analysis of main stilbenes and flavones in different parts (fronds, rhizomes, and frond bases) ofM. struthiopteris. The chromatographic separation was performed on a Kromasil C18 column (4.6 mm × 250 mm, 5 μm) with the mobile phase of MeOH-H2O (including 0.1% phosphoric acid) using a gradient elution at the flow rate of 1.0 mL min−1and UV detection at 295 nm. The method was validated by specificity, linearity, accuracy (recovery), and precision tests (repeatability, intra- and interday). For all the six compounds, the linear regression coefficients ranged from 0.9958 to 0.9998 within the test ranges; intra- and interday precisions were<2% and the mean recoveries ranged from 98.09 to 103.56%. The amount of these compounds in the frond bases was almost the same as in the rhizomes but much higher than that in the fronds. The results indicate that the HPLC method developed was appropriate for the analysis of the six compounds in different parts (fronds, rhizomes, and frond bases) ofM. struthiopteris.

scholarly journals Stability indicating RP-HPLC method for the estimation of flucloxacillin sodium in a tablet dosage form

2020 ◽  
Vol 1 (4) ◽  
pp. 95-100
Author(s):  
Sonalika Patro ◽  
S. Harshith Kumar ◽  
M. Barath kumar ◽  
E. Masthaniah ◽  
K. Sairam ◽  
...  
Keyword(s):  
Mobile Phase ◽  
Dosage Form ◽  
Theoretical Plate ◽  
Hplc Method ◽  
Rp Hplc ◽  
Ich Guidelines ◽  
System Suitability ◽  
Precision Study ◽  
Tablet Dosage

A Simple, accurate and precise method was developed and validated for the determination of flucloxacillin sodium in its tablet dosage form. The separation was eluted on xterra c18 column (4.6x150mm, 5micron) using a mixture of octane buffer and methanol as mobile phase in a ratio of (30:70) which was pumped through column at a flow rate of  1ml/min. Optimised wavelength for flucloxacillin was 237nm, the retention time was 2.305minutes and the percentage purity was found to be 98.14%. System suitability parameters such as theoretical plate and tailing factor for flucloxacillin sodium was found to be 2991.64 and 1.90 respectively, the proposed method was validated as per ICH guidelines (ICH, Q2 AND (R1)) the method was found to be linear at the concentration range of 20-100µg/ml and the correlation coefficient (r2) value was found to be 0.9994 percentage RSD for precision was 0.9% and percentage RSD for ruggedness was 0.5%. The precision study was precise, robust and repeatable. The LOD and LOQ values are 2.98 and 9.98 respectively. Hence the suggested RP-HPLC method can be used for routine analysis for flucloxacillin sodium in tablet dosage form.

scholarly journals Development and Validation of Estimation of Genotoxic Impurity (Hydroxylamine Hydrochloride content) in Leflunomide by using RP-HPLC technique

2021 ◽  
Vol 37 (02) ◽  
pp. 493-498
Author(s):  
Mohan Bhatale ◽  
Neelakandan Kaliyaperumal ◽  
Gopalakrishnan Mannathusamy ◽  
Gurunathan Ramalingam
Keyword(s):  
Mobile Phase ◽  
High Performance ◽  
Hydroxylamine Hydrochloride ◽  
High Performance Liquid Chromatographic ◽  
Hplc Method ◽  
Intermediate Precision ◽  
Rp Hplc ◽  
System Suitability ◽  
Development And Validation ◽  
Liquid Chromatographic

A simple, selective, linear having accuracy and specific of reverse phase high-performance liquid chromatographic (RP-HPLC) method for determination of Genotoxic impurity Hydroxylamine Hydrochloride of drug Leflunomide is reported.The separation and analysis were done on YMC Triart C18 (4.6 mm x 150 mm), having particle size 3.0 μm. KH2PO4 in 2000 mL of purified water and 2 mL triethylamine with pH 2.5 with phosphoric acid is mobile phase-A while acetonitrile is mobile Phase-B with gradient program. The elution achieved with 1.50 mL/min flow rate and using UV detection at 230 nm wavelength. Selected column oven temperature is 45°C and auto sampler 5°C respectively. In this method linearity and accuracy of Hydroxylamine HCl covered with specification limit of LOQ to 150 % (i.e.3 to 23 ppm). The observed correlation coefficient is 0.99965 and recovery in between 99.07 to 114.94. In method precision (ie.repeatability) and intermediate precision (IP) observed % RSD of six spiked test preparation is below 5.0 %. The standard and sample were stable for 3 days when stored at 2 to 8°C temperature. In robustness studies system suitability parameters ie tailing factor, theoretical plates and %RSD does not show significant changes. The present RP-HPLC method is selective, robust, linear, and precise for detection of Hydroxylamine HCl.

scholarly journals Comparative Study on Quantification of Total Catechins Using UV-Vis Spectrophotometric Method and High Performance Liquid Chromatography Techniques

2021 ◽  
Vol 37 (1) ◽  
pp. 136-142
Author(s):  
Sitharanjan Kalidass ◽  
Karuppannau Daiyarvijaya ◽  
Rajagopal Raj Kumar
Keyword(s):  
Quantitative Analysis ◽  
Spectrophotometric Method ◽  
Mobile Phase ◽  
High Performance ◽  
Cost Effective ◽  
Hplc Method ◽  
Good Resolution ◽  
Huge Number ◽  
Automated Hplc

Bioavailability of catechinsin wider range of plants was established earlier and it’s utility as medicine against cardiovascular disease, cancer, etc. were also demonstrated. Recent techniques in relation to quantitative analysis of total catechins seem to be laborious and time consuming process to handle huge number of samples. Established spectrophotometry and HPLC methods developed earlier for quantitative determination of total catechins in tea extracts were compared in the present study.UV-Vis spectrophotometric method was adopted to monitor the absorbance at 500 nm of the reaction mixture (catechins and vanillin-H2SO4reagents). Hewlett Packard automated HPLC was used and equipped with Phenomenex Luna 5  phenyl-hexyl column fitted with a Phenomenex guard column. Binary elution was carried out using Mobile phase A (acetic acid and acetonitrile) and Mobile phase B (acetonitrile). Method adopted showed a good resolution of catechin fractions and was found to be accurate for the quantification of total catechins (sum of individual catechins). Results of the both the methods are comparable and variation amongst the two methods ranged between -3.59 and 2.79% among the clones and varied with seasons.As expected UPASI released tea clones exhibited variations in their bioavailability. Lean season edge over the cropping period sampling in terms of total catechins. Results obtained from both the methods are comparable. Two methods can be used for the routine quantitative analysis of total catechins; however, spectrophotometric method found to be simple, rapid and cost effective than that of HPLC method unless individual catechins composition is warranted.

scholarly journals A 100% Water Mobile Phase HPLC-PDA Analysis of Selected Neonicotinoid Insecticides

2014 ◽  
Vol 10 (9) ◽  
pp. 3127-3132
Author(s):  
Naoto Furusawa
Keyword(s):  
Mobile Phase ◽  
Detection Limits ◽  
Hplc Method ◽  
Array Detector ◽  
Acceptance Criteria ◽  
Chromatographic Separations ◽  
Neonicotinoid Insecticides ◽  
Photodiode Array Detector ◽  
System Suitability ◽  
Photodiode Array

This paper describes a reserved-phase HPLC method for detecting frequently-used neonicotinoid insecticides, acetamiprid (ATP) and imidacloprid (ICP), using an isocratic 100 % water mobile phase.  Chromatographic separations were performed an Inertsil® WP300 C4 with water mobile phase and a photodiode-array detector.  The total run time was < 7 min.  The system suitability was well within the international acceptance criteria.  The detection limits were 0.013 μg ml-1 for ATP and 0.015 μg ml-1 for ICP, respectively.  A harmless HPLC method for simultaneous detecting ATP and ICP was developed and may be further applied to the quantification in foods.  

Method Development and Validation of Degradation Studies of Lenalidomide by RP-HPLC

2021 ◽  
pp. 4281-4286
Author(s):  
Punna Venkateshwarlu ◽  
Mehul M. Patel
Keyword(s):  
Retention Time ◽  
Mobile Phase ◽  
Method Development ◽  
Hplc Method ◽  
Mobile Phase Flow Rate ◽  
Pharmaceutical Dosage Forms ◽  
Rp Hplc ◽  
System Suitability ◽  
Wide Range

A simple, accurate, RP HPLC method was developed by this study determination of lenalidomide. This method is developed by Shimadzu LC -2010 HT by using C18 (250 X 4.6 X mm X 5µ) column in solvents Phosphate buffer: Acetonitrile (55:45) v/v as mobile phase and the temperature was maintained at 25°C. The mobile phase flow rate 1ml/min was pumped and sample wavelength was detected at 242nm by ultraviolet -visible spectrophotometer. The retention time was found 2.5 min. The number of theoretical plates and tailing factor for lenalidomide was observed 16199.817 (NLT 2000) and 1.128 (NMT 2). The method was validated for analytical standards such as linearity, accuracy, precision, system suitability and robustness. LOD and LOQ values obtained from regression of lenalidomide 0.058 and 0.174µg/ml. The regression equation of validated method for lenalidomide is Y=5223x+183075. In wide range of 25 to 150 (µg/ml) the linearity was observed. The method was validated and a recovery study indicates accuracy of this method. The Retention time less compared to established methods. The method was validated by determining its accuracy, precision and system suitability. The results of the study showed that the proposed RP-HPLC method is simple, rapid, precise and accurate, which is useful for the routine determination of Lenalidomide in bulk drug and in its pharmaceutical dosage forms.

A SIMPLE AND RAPID HPLC METHOD FOR ESTIMATION OF DIMETHICONE FROM FORMULATIONS

INDIAN DRUGS ◽  
2013 ◽  
Vol 50 (03) ◽  
pp. 26-29
Author(s):  
J. J Jadhav ◽  
◽  
S Mungekar ◽  
J. V. Velada ◽  
H. A. Doshi ◽  
...  
Keyword(s):  
Mobile Phase ◽  
High Performance ◽  
Dosage Forms ◽  
Hplc Method ◽  
Normal Phase ◽  
Hplc Separation ◽  
Tablet Dosage ◽  
Refractive Index Detector ◽  
Isocratic Mode

A simple, sensitive, precise and specific normal phase high performance liquid chromatography (HPLC) method was developed and validated for the determination of dimethicone from tablet dosage forms. It was found that the excipients used in the tablet dosage form did not interfere in the quantification of dimethicone. The HPLC separation was carried out by normal phase chromatography on Princeton Sphere Cyano, 250 x 4.6mm, 5µ with a mobile phase composed of hexane : ethanol : ethyl acetate (80:20:0.2) in isocratic mode at a flow rate of 0.5mL/min. Dimethicone was quantified using a refractive index detector. The calibration curve for dimethicone was linear from 1.75 to 3.25 mg/mL. The inter-day and intra-day precisions were found to be within limits. The proposed method has adequate sensitivity, reproducibility and specificity for the determination of dimethicone from tablet dosage forms.

Stereospecific Determination of Sertraline and its Impurities in Bulk Drug Using Cyclodextrins as a Chiral Selector

2020 ◽  
Vol 16 (7) ◽  
pp. 823-830 ◽  
Author(s):  
Navjot Kaur Sandhu ◽  
Durga Das Angehore ◽  
Neeraj Upmanyu ◽  
Pawan K. Porwal
Keyword(s):  
Mobile Phase ◽  
High Performance ◽  
Method Development ◽  
High Performance Liquid Chromatographic ◽  
Hplc Method ◽  
Related Substances ◽  
System Suitability ◽  
Bulk Drug ◽  
Liquid Chromatographic

Background: Sertraline Hydrochloride, an oral anti-depressant, has two chiral centers and its cis enantiomers and trans diasteromers are defined as related substances by United State Pharmacopoeia and British Pharmacopoeia. Introduction: A selective, stereospecific and economical high performance liquid chromatographic (HPLC) method was developed for the determination of sertraline enantiomeric forms. The HPLC-UV method was developed and optimized in the terms of system suitability parameters. Methods: The elution was made using a mixture of -cyclodextrin (-CD) and hydroxypropyl - cyclodextrin (HP -CD). Analysis was performed on a Zorbax SB C-18 column (250 x 4.6mm), 5μ with the mobile phase consisting of 170mM KH2PO4 containing -CD and HP -CD (pH: 3.0 with dil. H3PO4) and acetonitrile (75:25, v/v). Flow rate was kept at 1.0mL/min and the detection was carried out by UV at 220nm. Enantio-separation for sertraline was carried out using two different CDs (β-CD and HP β- CD) at different concentrations in the mobile phase. Results: Complete resolution of all the four isomers was achieved using 9mM β-CD and 15mM HP β- CD in the mobile phase. The development was optimized using central composite quadric model, where concentration of -CD and HP -CD were varied and resolution between trans diastereomeric impurities was calculated as a response. Conclusion: Resolution between any pair of isomers was found to be more than 2. Method development and optimization leading to the best resolution between the isomers of sertraline is described in detail.

scholarly journals Development and Validation of RP-HPLC Method for the Determination of Adefovir Dipivoxil in Bulk and in Pharmaceutical Formulation

2009 ◽  
Vol 6 (2) ◽  
pp. 469-474 ◽  
Author(s):  
Zaheer Ahmed ◽  
B. Gopinath ◽  
A. Sathish Kumar Shetty ◽  
B. K. Sridhar
Keyword(s):  
Flow Rate ◽  
Phosphate Buffer ◽  
Mobile Phase ◽  
Adefovir Dipivoxil ◽  
Hplc Method ◽  
Pharmaceutical Formulation ◽  
Rp Hplc ◽  
System Suitability ◽  
Development And Validation

A rapid and sensitive RP-HPLC method with UV detection (262 nm) for routine analysis of adefovir dipivoxil in bulk and in pharmaceutical formulation was developed. Chromatography was performed with mobile phase containing a mixture of acetonitrile and phosphate buffer (50:50, v/v) with flow rate 1.0 mL min-l. In the range of 5.0-100 µg/mL, the linearity of adefovir dipivoxil shows a correlation co-efficient of 0.9999. The proposed method was validated by determining sensitivity accuracy, precision, robustness stability, specificity, selectivity and system suitability parameters.

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