Simple high-performance liquid chromatographic column-switching technique for the on-line immunoaffinity extraction and analysis of flunitrazepam and its main metabolites in urine

Deinl, Ingo; Angermaier, L; Franzelius, Cornelia; Machbert, G.

doi:10.1016/s0378-4347(97)00429-5

Cited by 24 publications

(8 citation statements)

References 13 publications

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“…Sample preparation techniques involve SPE [36,40], automated on-line SPE [34], SPME [23,24,33], on-line dialysis [37,38], direct LC analysis using restricted access extraction materials [43,44], column switching techniques [45][46][47][48], as well as liquid phase microextraction [32].…”

Section: Introductionmentioning

confidence: 99%

Development of a validated HPLC method for the determination of four 1,4‐benzodiazepines in human biological fluids

Samanidou

Pechlivanidou

Papadoyannis³

2007

J of Separation Science

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A simple and sensitive HPLC method was developed and validated for the determination of four frequently prescribed 1,4-benzodiazepines: alprazolam (ALP), bromazepam (BRZ), diazepam (DZP), and flunitrazepam (FNZ). Separation was achieved on an Inertsil C8 analytical (250 mm x 4 mm, 5 microm) column, after selective extraction of benzodiazepine drugs from biological matrices by means of SPE. Isocratic elution was performed with a mobile phase consisting of CH3COONH4, 0.05 M CH3OH, and CH3CN (33:57:10 by volume). Quantification was performed at 240 nm with mefenamic acid (6 ng/microL) as the internal standard. DSC-18 Supelco cartridges provided high absolute recoveries (81-115%). The developed method was fully validated in terms of selectivity, linearity, accuracy, precision, stability, and sensitivity. Repeatability (n = 8) and between-day precision (n = 8) revealed RSD <12%. Recoveries from biological samples ranged from 81.2 to 115%. The detection limit of the method was calculated as 3.3-10.2 ng in blood plasma and 2.6-12.6 ng in urine for 20 microL injection volume. The method was applied to spiked biological matrices. Moreover, the method was applied to real samples of urine after an oral administration.

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Section: Introductionmentioning

confidence: 99%

Development of a validated HPLC method for the determination of four 1,4‐benzodiazepines in human biological fluids

Samanidou

Pechlivanidou

Papadoyannis³

2007

J of Separation Science

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“…The choice of detectors offers several possibilities among which are the coupling of HPLC/UV or HPLC/DAD [13][14][15][16][17][18][19][20][21][22][23][24][25][26][27], and more recently the coupling of HPLC/MS which offers interesting selectivity and specificity [28].…”

Section: Introductionmentioning

confidence: 99%

Rapid analysis of benzodiazepines in whole blood by high-performance liquid chromatography: use of a monolithic column

Bugey¹,

Staub²

2004

Journal of Pharmaceutical and Biomedical Analysis

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“…Simultaneous determination of flunitrazepam and its metabolites in human urine by UV absorbance was published by Deinl et al, (1997). Some assays involving protein analytes have utilized immunoassay coupled with size exclusion (Riggin et al, 1993) or ion-exchange chromatography (Janis and Regnier, 1988;Janis et al, 1989).…”

Section: Pre-column Immunoassays Coupled To Lcmentioning

confidence: 99%

Coupling immunoassays with chromatographic separation techniques

Tang

Karnes

2000

Biomed. Chromatogr.

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Coupling immunoassays with HPLC separation techniques is becoming increasingly useful in the analysis of biological and nonbiological samples of both large and small molecules. This is because it provides both sensitivity and selectivity for molecular analysis at relatively low cost, low maintenance and with excellent potential for automation. This paper reviews application of this hyphenated approach both in the pre-column immunoextraction and post-column immunodetection modes. Systems in which immunoassays are interfaced to chromatographic separations in order to separate bound and free fractions of the immunoassay will not be included since these systems do not provide the enhanced selectivity common to hyphenated systems. Post-column immunodetection is based on various immunoassay formats such as direct detection, one-site, competitive and sandwich immunoassays. Homogeneous immunodetectors are more convenient than heterogeneous immunodectors since there are no separation and column regeneration steps involved in homogeneous immunoassays. On the other hand, heterogeneous immunoassays are generally more sensitive than homogeneous immunoassays since interfering substances are removed prior to immunodetection. Advantages and limitations for the various approaches will be discussed.

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Simple high-performance liquid chromatographic column-switching technique for the on-line immunoaffinity extraction and analysis of flunitrazepam and its main metabolites in urine

Cited by 24 publications

References 13 publications

Development of a validated HPLC method for the determination of four 1,4‐benzodiazepines in human biological fluids

Development of a validated HPLC method for the determination of four 1,4‐benzodiazepines in human biological fluids

Rapid analysis of benzodiazepines in whole blood by high-performance liquid chromatography: use of a monolithic column

Coupling immunoassays with chromatographic separation techniques

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