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Development, validation and transfer of a hydrophilic interaction liquid chromatography/tandem mass spectrometric method for the analysis of the tobacco‐specific nitrosamine metabolite NNAL in human plasma at low picogram per milliliter concentrations
Author(s) -
Pan Jiongwei,
Song Qi,
Shi Haihong,
King Melissa,
Junga Heiko,
Zhou Shaolian,
Naidong Weng
Publication year - 2004
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1656
Subject(s) - chemistry , chromatography , metabolite , hydrophilic interaction chromatography , liquid chromatography–mass spectrometry , extraction (chemistry) , liquid–liquid extraction , detection limit , tandem mass spectrometry , solid phase extraction , mass spectrometry , sample preparation , high performance liquid chromatography , biochemistry
A highly sensitive bioanalytical method based on a simple liquid/liquid extraction and hydrophilic interaction liquid chromatography with tandem mass spectrometry (HILIC/MS/MS) analysis has been developed, validated and transferred for the determination of 4‐(methylnitrosamino)‐1‐(3‐pyridyl)‐1‐butanol (NNAL), a tobacco‐specific nitrosamine metabolite. Deuterated NNAL (NNAL‐d 4 ) was synthesized and used as the internal standard. This method can be used for the analysis of free and total NNAL (free NNAL plus NNAL‐gluc) in K 3 ‐EDTA human plasma. Free NNAL and NNAL‐d 4 are extracted from human plasma by liquid/liquid extraction. To analyze for total NNAL and the internal standard, a separate aliquot of the K 3 ‐EDTA human plasma is treated with β ‐glucuronidase to deconjugate the NNAL‐gluc; the total NNAL and internal standard are then extracted using liquid/liquid extraction. After drying down under nitrogen, the residue is reconstituted with acetonitrile and analyzed using positive ion electrospray and HILIC/MS/MS at a flow rate of 1.0 mL/min. The chromatographic run time is 1.0 min per injection, with retention time for both NNAL and NNAL‐d 4 of 0.75 min with a capacity factor (k') of 2. The standard curve range for this assay is from 5.00–1000 pg/mL for both free and total NNAL, using a total plasma sample volume of 1.0 mL. The interday precision and accuracy of the quality control (QC) samples demonstrated <7.6% relative standard deviation (RSD) and <3.3% relative error (RE) for free NNAL. For total NNAL, the interday precision and accuracy of the QC samples demonstrated <11.7% RSD and <2.8% RE. Optimization of enzyme hydrolysis of NNAL‐gluc is discussed in detail. The overall recoveries for free and total NNAL and IS were 68.2 and 71.5% (free) and 70.7 and 65.5% (total). No adverse matrix effects were noticed for this assay. Copyright © 2004 John Wiley & Sons, Ltd.