[Application Note] INTRODUCTION The determination or confirmation of the cause of a suspected poisoning is necessary for the rapid and effective treatment of patients admitted to a hospital after traumatic events such as attempted suicide, accidental poisoning or an adverse drug reaction. Various analytical approaches are possible, which include GC, GC/MS, LC/MS, immunoassay, or LC/UV with the use of UV spectra libraries. LC/UV is a popular analytical technique as it is often found as standard equipment in many laboratories and patient samples do not routinely require derivatization after extraction, thereby reducing both cost and time. Additionally, the methodology can be semi quantitative. The identification of xenobiotics combines both LC retention time and UV spectra, with a further comparison to a compound library. This type of analytical application is operated routinely in many laboratories with the use of Waters Alliance ® systems, incorporating a method developed by ToxLab in Paris 1 . Some potential limitations of this particular application relate to chromatographic interferences which impact the UV spectra, leading to inferior library matches. The introduction of UPLC ® systems has provided dramatic improvements in the resolution of chromatographic peaks, analytical sensitivity and reproducibility of the retention times. Transferring the application from HPLC to UPLC will therefore improve the identification of any unknown compounds in a shorter time. The compound library that has been created, contains separate UV spectra for 612 compounds along with their relative retention times for the UPLC method as described hereafter. All chromato- grams and tables shown in this document are kindly provided, courtesy of Mrs. Camille Chatenay. TOXICOLOGY SCREENING BY UPLC/PDA IN COMBINATION WITH AN EXTENSIVE COMPOUND LIBRARY Camille Chatenay, 1,2 Fabien Bévalot, 1,2 Cyril Mounier, 1 Jean Michel Prévosto 1 1 Hôpital d’Instruction des Armées Desgenettes, Lyon (France), 2 Laboratoire LUMTOX, Lyon (France) EXPERIMENTAL LC Conditions LC System: Waters ACQUITY UPLC ® System Column: ACQUITY UPLC BEH C 18 Column 2.1 x 150 mm, 1.7 μm Column Temp: 50 ˚C Flow Rate: 450 μL/min Mobile Phase A: Acetonitrile/ Ammonium acetate buffer 5 mM pH 3.8 (15/85) Mobile Phase B: Acetonitrile Gradient: Time (min) % A % B 0.00 100 0.0 1.40 100 0.0 2.80 89.0 11.0 11.00 25.0 75.0 11.20 25.0 75.0 12.00 100 0.0 15.00 100 0.0 Detector: Waters UPLC PDA detector 2 Wavelength range: 200-400 nm, 40 points/second Sample preparation: Sample volume: 1 mL Extraction: LLE with Toxitube A* Internal standard: 20 µL flurazepam at (reference peak) 0.05 mg/mL Reconstitution: 80 µL volume Injected volume: 5 µL * Varian Inc. Waters ACQUITY UPLC system
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Toxicology Screening by UPLC/PDA in Combination with an ...The test mixture is composed of the following compounds: Venlafaxine, Theophylline, Amisulpride, Mirtazapine, Trifluoperazine,
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[Application Note]
INT RODUCT ION
The determination or confirmation of the cause of a suspected
poisoning is necessary for the rapid and effective treatment of
patients admitted to a hospital after traumatic events such as
attempted suicide, accidental poisoning or an adverse drug reaction.
Various analytical approaches are possible, which include GC,
GC/MS, LC/MS, immunoassay, or LC/UV with the use of UV spectra
libraries.
LC/UV is a popular analytical technique as it is often found as
standard equipment in many laboratories and patient samples
do not routinely require derivatization after extraction, thereby
reducing both cost and time. Additionally, the methodology can
be semi quantitative. The identification of xenobiotics combines
both LC retention time and UV spectra, with a further comparison
to a compound library. This type of analytical application is
operated routinely in many laboratories with the use of Waters
Alliance® systems, incorporating a method developed by ToxLab
in Paris1. Some potential limitations of this particular application
relate to chromatographic interferences which impact the UV
spectra, leading to inferior library matches.
The introduction of UPLC® systems has provided dramatic
improvements in the resolution of chromatographic peaks,
analytical sensitivity and reproducibility of the retention times.
Transferring the application from HPLC to UPLC will therefore
improve the identification of any unknown compounds in a shorter
time.
The compound library that has been created, contains separate
UV spectra for 612 compounds along with their relative retention
times for the UPLC method as described hereafter. All chromato-
grams and tables shown in this document are kindly provided,
courtesy of Mrs. Camille Chatenay.
T OX ICO LOGY SC R E ENING BY U P LC / P DA IN COMBINAT ION W IT H AN E X T ENSIV E COM P OUND L IB RA RY
Camille Chatenay,1,2 Fabien Bévalot,1,2 Cyril Mounier,1 Jean Michel Prévosto1