Development and validation of a liquid chromatography-tandem mass spectrometry method for the determination of nicotine and its metabolites in placenta and umbilical cord.
de Castro, Ana
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PANIAGUA-GONZÁLEZ. L., JIMÉNEZ-MORIGOSA, C., LENDOIRO, E., CONCHEIRO, M., CRUZ, A., LÓPEZ-RIVADULLA, M. and DE CASTRO, A. 2018. Development and validation of a liquid chromatography-tandem mass spectrometry method for the determination of nicotine and its metabolites in placenta and umbilical cord. Drug testing and analysis [online], 10(8), pages 1305-1314. Available from: https://doi.org/10.1002/dta.2381
Tobacco exposure during pregnancy is associated with obstetric and fetal complications. We developed and validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine nicotine, cotinine, and hydroxycotinine (OH-cotinine) in placenta (PL) and umbilical cord (UC). Specimens were homogenized in water, followed by solid-phase extraction. Chromatographic separation was performed using an Atlantis® HILIC Silica column. Detection was accomplished in electrospray in positive mode. Method validation included: linearity (5 to 1000 ng/g), accuracy (86.9 to 105.2% of target concentration in PL, and 89.1 to 105.0% in UC), imprecision (6.8 to 11.8% in PL, and 7.6 to 12.2% in UC), limits of detection (2 ng/g for cotinine and OH-cotinine, and 5 ng/g for nicotine) and quantification (5 ng/g), selectivity (no endogenous or exogenous interferences), matrix effect (-34.1 to -84.5% in PL, %CV = 9.1-24.0%; -18.9 to -84.7% in UC, %CV = 10.2-23.9%), extraction efficiency (60.7 to 131.5% in PL, and 64.1 to 134.2% in UC), and stability 72 h in the autosampler (<11.5% loss in PL, and < 13% loss in UC). The method was applied to 14 PL and UC specimens from tobacco users during pregnancy. Cotinine (6.8-312.2 ng/g in PL; 6.7-342.3 ng/g in UC) was the predominant analyte, followed by OH-cotinine (<LOQ-80.2 ng/g in PL; <LOQ-80.5 ng/g in UC) and nicotine (5.7-63.7 ng/g in PL; 5.1-63.3 ng/g in UC). This method will be applied to more than 150 specimens collected from a wide clinical study to evaluate the usefulness of maternal hair, meconium, placenta and umbilical cord compared to the maternal interview to detect in utero drug exposure.