Development and validation of a high-performance liquid chromatography-fluorescence detection method for the accurate quantification of colistin in human plasma

Divyabharathi Chepyala, I. Lin Tsai, Hsin Yun Sun, Shu Wen Lin, Ching Hua Kuo

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Recently, colistin has become one of the most important drugs for treating infections caused by multidrug-resistant Gram-negative bacteria. Therapeutic drug monitoring is recommended to ensure the safety and efficacy of colistin and to improve clinical outcomes. This study developed an accurate and sensitive high-performance liquid chromatography-fluorescence detection (HPLC-FLD) method for the quantification of colistin in human plasma. The sample preparation included protein precipitation using trichloroacetic acid (TCA) and methanol, followed by in-solid phase extraction (In-SPE) derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl). A Poroshell 120 EC-C18 2.1×100mm (2.7μm) column was used in the HPLC method with a mobile phase composed of acetonitrile (ACN), tetrahydrofuran (THF), and deionized (DI) water (82%, 2%, 16% (v/v), respectively). Polymyxin B1 was used as the internal standard. The total analysis time was 22min under optimal separation conditions. The HPLC-FLD method was validated over a therapeutic range of 0.3-6.0μgmL-1. The intra-day and inter-day precisions for colistin A and colistin B were below 9.9% and 4.5% relative standard deviations, respectively. The accuracy test results were between 100.2 and 118.4%. The extraction recoveries were between 81.6 and 94.1%. The method was linear over the test range, with a 0.9991 coefficient of determination. The limit of detection was 0.1μgmL-1. The validated HPLC-FLD method was successfully applied to quantify the colistin concentrations in 2 patient samples for therapeutic drug monitoring.

Original languageEnglish
Pages (from-to)48-54
Number of pages7
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume980
DOIs
Publication statusPublished - Feb 1 2015
Externally publishedYes

Fingerprint

Plasma (human)
Colistin
High performance liquid chromatography
Fluorescence
High Pressure Liquid Chromatography
Drug Monitoring
Pharmaceutical Preparations
Trichloroacetic Acid
Monitoring
Deionized water
Solid Phase Extraction
Gram-Negative Bacteria
Methanol
Limit of Detection
Bacteria
Safety
Recovery
Water
Infection
Proteins

Keywords

  • Colistin
  • FMOC-Cl
  • HPLC-FLD
  • Human plasma
  • Therapeutic drug monitoring

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Cell Biology
  • Clinical Biochemistry
  • Medicine(all)

Cite this

@article{32bec6337b674dd5a094343867158e06,
title = "Development and validation of a high-performance liquid chromatography-fluorescence detection method for the accurate quantification of colistin in human plasma",
abstract = "Recently, colistin has become one of the most important drugs for treating infections caused by multidrug-resistant Gram-negative bacteria. Therapeutic drug monitoring is recommended to ensure the safety and efficacy of colistin and to improve clinical outcomes. This study developed an accurate and sensitive high-performance liquid chromatography-fluorescence detection (HPLC-FLD) method for the quantification of colistin in human plasma. The sample preparation included protein precipitation using trichloroacetic acid (TCA) and methanol, followed by in-solid phase extraction (In-SPE) derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl). A Poroshell 120 EC-C18 2.1×100mm (2.7μm) column was used in the HPLC method with a mobile phase composed of acetonitrile (ACN), tetrahydrofuran (THF), and deionized (DI) water (82{\%}, 2{\%}, 16{\%} (v/v), respectively). Polymyxin B1 was used as the internal standard. The total analysis time was 22min under optimal separation conditions. The HPLC-FLD method was validated over a therapeutic range of 0.3-6.0μgmL-1. The intra-day and inter-day precisions for colistin A and colistin B were below 9.9{\%} and 4.5{\%} relative standard deviations, respectively. The accuracy test results were between 100.2 and 118.4{\%}. The extraction recoveries were between 81.6 and 94.1{\%}. The method was linear over the test range, with a 0.9991 coefficient of determination. The limit of detection was 0.1μgmL-1. The validated HPLC-FLD method was successfully applied to quantify the colistin concentrations in 2 patient samples for therapeutic drug monitoring.",
keywords = "Colistin, FMOC-Cl, HPLC-FLD, Human plasma, Therapeutic drug monitoring",
author = "Divyabharathi Chepyala and Tsai, {I. Lin} and Sun, {Hsin Yun} and Lin, {Shu Wen} and Kuo, {Ching Hua}",
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TY - JOUR

T1 - Development and validation of a high-performance liquid chromatography-fluorescence detection method for the accurate quantification of colistin in human plasma

AU - Chepyala, Divyabharathi

AU - Tsai, I. Lin

AU - Sun, Hsin Yun

AU - Lin, Shu Wen

AU - Kuo, Ching Hua

PY - 2015/2/1

Y1 - 2015/2/1

N2 - Recently, colistin has become one of the most important drugs for treating infections caused by multidrug-resistant Gram-negative bacteria. Therapeutic drug monitoring is recommended to ensure the safety and efficacy of colistin and to improve clinical outcomes. This study developed an accurate and sensitive high-performance liquid chromatography-fluorescence detection (HPLC-FLD) method for the quantification of colistin in human plasma. The sample preparation included protein precipitation using trichloroacetic acid (TCA) and methanol, followed by in-solid phase extraction (In-SPE) derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl). A Poroshell 120 EC-C18 2.1×100mm (2.7μm) column was used in the HPLC method with a mobile phase composed of acetonitrile (ACN), tetrahydrofuran (THF), and deionized (DI) water (82%, 2%, 16% (v/v), respectively). Polymyxin B1 was used as the internal standard. The total analysis time was 22min under optimal separation conditions. The HPLC-FLD method was validated over a therapeutic range of 0.3-6.0μgmL-1. The intra-day and inter-day precisions for colistin A and colistin B were below 9.9% and 4.5% relative standard deviations, respectively. The accuracy test results were between 100.2 and 118.4%. The extraction recoveries were between 81.6 and 94.1%. The method was linear over the test range, with a 0.9991 coefficient of determination. The limit of detection was 0.1μgmL-1. The validated HPLC-FLD method was successfully applied to quantify the colistin concentrations in 2 patient samples for therapeutic drug monitoring.

AB - Recently, colistin has become one of the most important drugs for treating infections caused by multidrug-resistant Gram-negative bacteria. Therapeutic drug monitoring is recommended to ensure the safety and efficacy of colistin and to improve clinical outcomes. This study developed an accurate and sensitive high-performance liquid chromatography-fluorescence detection (HPLC-FLD) method for the quantification of colistin in human plasma. The sample preparation included protein precipitation using trichloroacetic acid (TCA) and methanol, followed by in-solid phase extraction (In-SPE) derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl). A Poroshell 120 EC-C18 2.1×100mm (2.7μm) column was used in the HPLC method with a mobile phase composed of acetonitrile (ACN), tetrahydrofuran (THF), and deionized (DI) water (82%, 2%, 16% (v/v), respectively). Polymyxin B1 was used as the internal standard. The total analysis time was 22min under optimal separation conditions. The HPLC-FLD method was validated over a therapeutic range of 0.3-6.0μgmL-1. The intra-day and inter-day precisions for colistin A and colistin B were below 9.9% and 4.5% relative standard deviations, respectively. The accuracy test results were between 100.2 and 118.4%. The extraction recoveries were between 81.6 and 94.1%. The method was linear over the test range, with a 0.9991 coefficient of determination. The limit of detection was 0.1μgmL-1. The validated HPLC-FLD method was successfully applied to quantify the colistin concentrations in 2 patient samples for therapeutic drug monitoring.

KW - Colistin

KW - FMOC-Cl

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KW - Human plasma

KW - Therapeutic drug monitoring

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