Development and validation of a gradient HPLC method for the determination of insulin in rat plasma: application to pharmacokinetic study / Eda Frida Salleh

A simple, specific and sensitive reversed-phase high performance liquid chromatography method was developed to determine insulin in rat plasma. The method involves direct injection of rat plasma sample after deproteinization using acetonitrile - propanol (1:1). The mobile phase comprises a mixture o...

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Main Author: Salleh, Eda Frida
Format: Thesis
Language:English
Published: 2012
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Online Access:https://ir.uitm.edu.my/id/eprint/65561/1/65561.pdf
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spelling my-uitm-ir.655612022-11-07T07:10:58Z Development and validation of a gradient HPLC method for the determination of insulin in rat plasma: application to pharmacokinetic study / Eda Frida Salleh 2012 Salleh, Eda Frida Drugs and their actions A simple, specific and sensitive reversed-phase high performance liquid chromatography method was developed to determine insulin in rat plasma. The method involves direct injection of rat plasma sample after deproteinization using acetonitrile - propanol (1:1). The mobile phase comprises a mixture of water and acetonitrile containing 0.1% v/v trifloroacetic acid (TFA) with gradient analysis by increasing the proportion of mobile phase of water from 25% v/v to 40% v/v within 20 minutes. A Phenomenon C18 column (5μm 4.6 x 150 mm) was used for the chromatographic separation which was delivered at a flow rate of 0.6 mL/min at 210 nm wavelength. Under these conditions, the method was validated with respect to linearity, recovery, specificity, accuracy, precision, and stability. The method was proven to be linear over the concentration range of 0.39 - 50.00 μg/ml with mean correlation coefficient of 0.9994. The mean extraction recovery was 97.4%, while the precision of within-day and between-day measurements was a <9%. The limit of detection (LOD) and limit of quantitation (LOQ) of the method were 0.13 μg/ml and 0.39 μg/ml, respectively. The short run time, specificity, sensitivity and reproducibility of this method were found to be satisfactory and thus suitable for routine determination of insulin in rat. 2012 Thesis https://ir.uitm.edu.my/id/eprint/65561/ https://ir.uitm.edu.my/id/eprint/65561/1/65561.pdf text en public degree Universiti Teknologi MARA (UiTM) Faculty of Pharmacy Abdul Hamid, Khuriah
institution Universiti Teknologi MARA
collection UiTM Institutional Repository
language English
advisor Abdul Hamid, Khuriah
topic Drugs and their actions
spellingShingle Drugs and their actions
Salleh, Eda Frida
Development and validation of a gradient HPLC method for the determination of insulin in rat plasma: application to pharmacokinetic study / Eda Frida Salleh
description A simple, specific and sensitive reversed-phase high performance liquid chromatography method was developed to determine insulin in rat plasma. The method involves direct injection of rat plasma sample after deproteinization using acetonitrile - propanol (1:1). The mobile phase comprises a mixture of water and acetonitrile containing 0.1% v/v trifloroacetic acid (TFA) with gradient analysis by increasing the proportion of mobile phase of water from 25% v/v to 40% v/v within 20 minutes. A Phenomenon C18 column (5μm 4.6 x 150 mm) was used for the chromatographic separation which was delivered at a flow rate of 0.6 mL/min at 210 nm wavelength. Under these conditions, the method was validated with respect to linearity, recovery, specificity, accuracy, precision, and stability. The method was proven to be linear over the concentration range of 0.39 - 50.00 μg/ml with mean correlation coefficient of 0.9994. The mean extraction recovery was 97.4%, while the precision of within-day and between-day measurements was a <9%. The limit of detection (LOD) and limit of quantitation (LOQ) of the method were 0.13 μg/ml and 0.39 μg/ml, respectively. The short run time, specificity, sensitivity and reproducibility of this method were found to be satisfactory and thus suitable for routine determination of insulin in rat.
format Thesis
qualification_level Bachelor degree
author Salleh, Eda Frida
author_facet Salleh, Eda Frida
author_sort Salleh, Eda Frida
title Development and validation of a gradient HPLC method for the determination of insulin in rat plasma: application to pharmacokinetic study / Eda Frida Salleh
title_short Development and validation of a gradient HPLC method for the determination of insulin in rat plasma: application to pharmacokinetic study / Eda Frida Salleh
title_full Development and validation of a gradient HPLC method for the determination of insulin in rat plasma: application to pharmacokinetic study / Eda Frida Salleh
title_fullStr Development and validation of a gradient HPLC method for the determination of insulin in rat plasma: application to pharmacokinetic study / Eda Frida Salleh
title_full_unstemmed Development and validation of a gradient HPLC method for the determination of insulin in rat plasma: application to pharmacokinetic study / Eda Frida Salleh
title_sort development and validation of a gradient hplc method for the determination of insulin in rat plasma: application to pharmacokinetic study / eda frida salleh
granting_institution Universiti Teknologi MARA (UiTM)
granting_department Faculty of Pharmacy
publishDate 2012
url https://ir.uitm.edu.my/id/eprint/65561/1/65561.pdf
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