A New, Rapid, Fully Automated Method for Determination of Fluconazole in Serum by Column-Switching Liquid Chromatography

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A New, Rapid, Fully Automated Method for Determination of Fluconazole in Serum by Column-Switching Liquid Chromatography. / Egle, Hannes; Trittler, Rainer; Kümmerer, Klaus.
in: Therapeutic Drug Monitoring, Jahrgang 26, Nr. 4, 01.08.2004, S. 425-431.

Publikation: Beiträge in ZeitschriftenZeitschriftenaufsätzeForschungbegutachtet

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@article{bc640e8e1c8e485fba69fb90e9d207a4,
title = "A New, Rapid, Fully Automated Method for Determination of Fluconazole in Serum by Column-Switching Liquid Chromatography",
abstract = "A sensitive and rapid HPLC assay for the determination of fluconazole in serum is described. HPLC-integrated sample preparation allows direct injection of serum samples without any pretreatment. The in-line extraction technique is carried out by automatically switching from the extraction column (Lichrospher{\textregistered} ADS C 8) to the analytic column (Nucleosil C 18). After 6 minutes the matrix passes the extraction column, and the retained analyte is quantitatively transferred to the analytic column, where separation by isocratic HPLC is performed. The extraction eluent is sodium dihydrogen phosphate buffer, pH 5.0 (50 mM), and the analytic eluent is acetonitrile/sodium dihydrogen phosphate buffer, pH 5.0 (50 mM) (26.8/73.2, vol/vol). Fluconazole is detected according to its absorption maximum at 210 nm. The lower limit of quantification (LLOQ) is 0.65 μg/mL, the limit of detection (LOD) is 0.2 μg/mL, and the quantification range is 0.65-23.3 μg/mL. The assay was precise with a between-run coefficient of variation of ≤ 5.59%. The within-run accuracy was 99.8% and 103.4%, and the between-run accuracy was 99.2% and 99.7%, respectively, for the concentrations 23.3 μg/mL and 1.3 μg/mL. The recovery was 78%. The described procedure allows sample cleanup and determination within 20 minutes, thereby facilitating drug monitoring in clinical routine. The method was applied successfully. ",
keywords = "Absorption, accuracy, ADS, ASSAY, Bioassay, buffer, chromatography, COEFFICIENT, COLUMN, column switching, column-switching, concentration, detection, determination, direct injection, DIRECT-INJECTION, DOSAGE, drug, drug monitoring, Drug-monitoring, EXTRACTION, FLUCONAZOLE, GERMANY, HPLC, HPLC ASSAY, human plasma, in-line extraction, INJECTION, INTEGRATED SAMPLE PREPARATION, liquid, liquid chromatography, LIQUID-CHROMATOGRAPHY, MALIGNANCY, MATRIX, Method, Monitoring, PH, pharmacokinetics, PRETREATMENT, QUANTIFICATION, recovery, RENAL REPLACEMENT THERAPY, sample preparation, SAMPLES, SEPARATION, Serum, SERUM SAMPLE, TANDEM MASS-SPECTROMETRY, USA, Chemistry",
author = "Hannes Egle and Rainer Trittler and Klaus K{\"u}mmerer",
year = "2004",
month = aug,
day = "1",
doi = "10.1097/00007691-200408000-00013",
language = "English",
volume = "26",
pages = "425--431",
journal = "Therapeutic Drug Monitoring",
issn = "0163-4356",
publisher = "Lippincott Williams and Wilkins",
number = "4",

}

RIS

TY - JOUR

T1 - A New, Rapid, Fully Automated Method for Determination of Fluconazole in Serum by Column-Switching Liquid Chromatography

AU - Egle, Hannes

AU - Trittler, Rainer

AU - Kümmerer, Klaus

PY - 2004/8/1

Y1 - 2004/8/1

N2 - A sensitive and rapid HPLC assay for the determination of fluconazole in serum is described. HPLC-integrated sample preparation allows direct injection of serum samples without any pretreatment. The in-line extraction technique is carried out by automatically switching from the extraction column (Lichrospher® ADS C 8) to the analytic column (Nucleosil C 18). After 6 minutes the matrix passes the extraction column, and the retained analyte is quantitatively transferred to the analytic column, where separation by isocratic HPLC is performed. The extraction eluent is sodium dihydrogen phosphate buffer, pH 5.0 (50 mM), and the analytic eluent is acetonitrile/sodium dihydrogen phosphate buffer, pH 5.0 (50 mM) (26.8/73.2, vol/vol). Fluconazole is detected according to its absorption maximum at 210 nm. The lower limit of quantification (LLOQ) is 0.65 μg/mL, the limit of detection (LOD) is 0.2 μg/mL, and the quantification range is 0.65-23.3 μg/mL. The assay was precise with a between-run coefficient of variation of ≤ 5.59%. The within-run accuracy was 99.8% and 103.4%, and the between-run accuracy was 99.2% and 99.7%, respectively, for the concentrations 23.3 μg/mL and 1.3 μg/mL. The recovery was 78%. The described procedure allows sample cleanup and determination within 20 minutes, thereby facilitating drug monitoring in clinical routine. The method was applied successfully.

AB - A sensitive and rapid HPLC assay for the determination of fluconazole in serum is described. HPLC-integrated sample preparation allows direct injection of serum samples without any pretreatment. The in-line extraction technique is carried out by automatically switching from the extraction column (Lichrospher® ADS C 8) to the analytic column (Nucleosil C 18). After 6 minutes the matrix passes the extraction column, and the retained analyte is quantitatively transferred to the analytic column, where separation by isocratic HPLC is performed. The extraction eluent is sodium dihydrogen phosphate buffer, pH 5.0 (50 mM), and the analytic eluent is acetonitrile/sodium dihydrogen phosphate buffer, pH 5.0 (50 mM) (26.8/73.2, vol/vol). Fluconazole is detected according to its absorption maximum at 210 nm. The lower limit of quantification (LLOQ) is 0.65 μg/mL, the limit of detection (LOD) is 0.2 μg/mL, and the quantification range is 0.65-23.3 μg/mL. The assay was precise with a between-run coefficient of variation of ≤ 5.59%. The within-run accuracy was 99.8% and 103.4%, and the between-run accuracy was 99.2% and 99.7%, respectively, for the concentrations 23.3 μg/mL and 1.3 μg/mL. The recovery was 78%. The described procedure allows sample cleanup and determination within 20 minutes, thereby facilitating drug monitoring in clinical routine. The method was applied successfully.

KW - Absorption

KW - accuracy

KW - ADS

KW - ASSAY

KW - Bioassay

KW - buffer

KW - chromatography

KW - COEFFICIENT

KW - COLUMN

KW - column switching

KW - column-switching

KW - concentration

KW - detection

KW - determination

KW - direct injection

KW - DIRECT-INJECTION

KW - DOSAGE

KW - drug

KW - drug monitoring

KW - Drug-monitoring

KW - EXTRACTION

KW - FLUCONAZOLE

KW - GERMANY

KW - HPLC

KW - HPLC ASSAY

KW - human plasma

KW - in-line extraction

KW - INJECTION

KW - INTEGRATED SAMPLE PREPARATION

KW - liquid

KW - liquid chromatography

KW - LIQUID-CHROMATOGRAPHY

KW - MALIGNANCY

KW - MATRIX

KW - Method

KW - Monitoring

KW - PH

KW - pharmacokinetics

KW - PRETREATMENT

KW - QUANTIFICATION

KW - recovery

KW - RENAL REPLACEMENT THERAPY

KW - sample preparation

KW - SAMPLES

KW - SEPARATION

KW - Serum

KW - SERUM SAMPLE

KW - TANDEM MASS-SPECTROMETRY

KW - USA

KW - Chemistry

UR - http://www.scopus.com/inward/record.url?scp=3343018904&partnerID=8YFLogxK

U2 - 10.1097/00007691-200408000-00013

DO - 10.1097/00007691-200408000-00013

M3 - Journal articles

VL - 26

SP - 425

EP - 431

JO - Therapeutic Drug Monitoring

JF - Therapeutic Drug Monitoring

SN - 0163-4356

IS - 4

ER -

DOI