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
Subjects:	Drugs and their actions
Online Access:	https://ir.uitm.edu.my/id/eprint/65561/1/65561.pdf
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id	my-uitm-ir.65561
record_format	uketd_dc
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
_version_	1783735559547518976

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

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