An optimized and validated RP-HPLC/UV detection method for simultaneous determination of all-trans-Retinol (Vitamin A) and α-Tocopherol (Vitamin E) in human serum: Comparison of different particulate reversed-phase HPLC columns

Khan, Abad; Khan, Muhammad Imran; Iqbal, Zafar; Shah, Yatrik M.; Ahmad, Lateef; Watson, David

doi:10.1016/j.jchromb.2010.07.009

Cited by 46 publications

(34 citation statements)

References 61 publications

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“…A short run time in the HPLC method always facilitate analysis of several samples per day as already published in some of the studies with HPLC methods, (13,(28)(29)(30). The run time of 2.5 min in this study also facilitates analysis of several hundred samples of TNF per day.…”

Section: Introductionsupporting

confidence: 53%

Sensitive and Rapid HPLC Quantification of Tenofovir from Hyaluronic Acid-Based Nanomedicine

Agrahari

Youan

2012

AAPS PharmSciTech

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Abstract. The purpose of this study was to develop and validate a rapid, sensitive, and specific reversedphase high-performance liquid chromatography method for the quantitative determination of native tenofovir (TNF) for various applications. Different analytical performance parameters such as linearity, precision, accuracy, limit of quantification (LOQ), limit of detection (LOD), and robustness were determined according to International Conference on Harmonization (ICH) guidelines. A Bridge™ C18 column (150×4.6 mm, 5 μm) was used as stationary phase. The retention time of TNF was 1.54±0.03 min (n=6). The assay was linear over the concentration range of 0.1-10 μg/mL. The proposed method was sensitive with LOD and LOQ values equal to 50 and 100 ng/mL, respectively. The method was accurate with percent mean recovery from 95.41% to 102.90% and precise as percent RSD (relative standard deviation) values for intra-day, and inter-day precision were less than 2%. This method was utilized for the estimation of molar absorptivity of TNF at 259 nm (ε 259 =12,518 L/mol/cm), calculated from linear regression analysis. The method was applied for determination of percentage of encapsulation efficiency (22.93±0.04%), drug loading (12.25±1.03%), in vitro drug release profile in the presence of enzyme (43% release in the first 3 h) and purification analysis of hyaluronic acid-based nanomedicine.

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Section: Introductionsupporting

confidence: 53%

Sensitive and Rapid HPLC Quantification of Tenofovir from Hyaluronic Acid-Based Nanomedicine

Agrahari

Youan

2012

AAPS PharmSciTech

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“…The liquid-liquid extraction internal standard, retinol acetate, was deemed a suitable internal standard for this assay because of its documented use as suitable internal standard in previous investigations involving fat soluble vitamins, including both retinol and the tocopherols [22][23][24]26,36].…”

Section: Internal Standardmentioning

confidence: 99%

“…Several methods have been developed for the simultaneous monitoring of tocopherol and retinol in human plasma [5][6][7][8][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27]29,30,[32][33][34][35][36][37]40,[43][44][45][46]. These methods have often featured chromatography run-times and flow rates requiring considerable consumption of time and/or solvents during sample analyses [16-18, 20,21,29,35,47].…”

Section: Introductionmentioning

confidence: 99%

Fluorometric Determination of Vitamin Constituents in Human Plasma Using Ultra Performance Liquid Chromatography

Bell¹

2012

J Chromatogr Sep Tech

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Purpose: A rapid, selective and sensitive ultra performance liquid chromatography method has been developed for the detection and quantification of several vitamins in human plasma.Methods: Alpha tocopherol, gamma tocopherol, and retinol are assayed using fluorescence detection. Excitation/emission wavelengths are 295 nm/330 nm and 325 nm/470 nm for the analysis of both tocopherols and retinol, respectively. Retinol acetate is employed as the internal standard. The reversed phase method incorporates gradient elution with a mobile phase consisting of methanol and acetonitrile. Separation of vitamin compounds is achieved using a bridged ethyl-hybrid C18 column. Results:The retention times for retinol, retinol acetate, gamma tocopherol and alpha tocopherol are 1.6 minutes, 1.8 minutes, 3.9 minutes and 4.3 minutes, respectively. The limits of quantification for retinol, gamma tocopherol and alpha tocopherol, were 0.02 μg/ml, 0.02 μg/ml, and 0.1 μg/ml, respectively. Conclusion:The assay method is suitable for the analysis of these vitamins in human plasma following the ingestion of foods fortified with these fat soluble vitamins.

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“…Availability of more than 1000 commercial stationary phases [2,3], makes column selection a crucial process. Therefore, it is not surprising that numerous papers [4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22] describing various attempts to achieve this are being continuously published.…”

Section: Introductionmentioning

confidence: 99%

Assessment of column selection systems using Partial Least Squares

Žuvela

Liu

Plenis

et al. 2015

Journal of Chromatography A

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An optimized and validated RP-HPLC/UV detection method for simultaneous determination of all-trans-Retinol (Vitamin A) and α-Tocopherol (Vitamin E) in human serum: Comparison of different particulate reversed-phase HPLC columns

Cited by 46 publications

References 61 publications

Sensitive and Rapid HPLC Quantification of Tenofovir from Hyaluronic Acid-Based Nanomedicine

Sensitive and Rapid HPLC Quantification of Tenofovir from Hyaluronic Acid-Based Nanomedicine

Fluorometric Determination of Vitamin Constituents in Human Plasma Using Ultra Performance Liquid Chromatography

Assessment of column selection systems using Partial Least Squares

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