Gas chromatography-mass spectrometry determination of nicotine and cotinine in urine: A study of the effect of passive smoking.

RATIONALE: Recent data suggest that passive smoking has a risk comparable to active smoking. Passive smoking is considered dangerous in children and is suspected as a cause of asthma. However, some reports are opposing such claims, indicating the need for solid results and large-scale studies. This scientific work aims to develop a method for the determination of nicotine (NCOT) and major nicotine's metabolite cotinine (COT) in urine samples, using gas chromatography-mass spectrometry (GC-MS). METHODS: Analysis was performed using a gas chromatograph Agilent Technologies 7890A with an MS 5975C inert XL, EI/CI MSD with Triple-Axis detector. For sample preparation, liquid-liquid extraction was applied after an optimization study with different extraction media. Eventually, 1 mL of dichloromethane was selected for the extraction of 0.5 mL of urine. Suitable chromatographic conditions were found for the rapid and accurate determination of NCOT and COT. Injection of 2 µL was performed using GC-MS, and selected ion monitoring (SIM) analysis was performed with the following ions (m/z): 162 (quantifier ion) and 84, 133, 161 qualifier ions for NCOT, and 176 (quantifier ion) and 98, 118, 119, 147 qualifier ions for COT. Nicotine-D4 (NCOT-D4) and cotinine-D3 (COT-D3) were used as internal standards with quantifier ions 101 and 166, respectively. The retention time (Rt) for NCOT was 7.557 min and 9.743 min for COT. RESULTS: The method was validated following international principles, assessing characteristics such as absolute recovery, carryover, linearity, specificity, selectivity, accuracy, precision, and stability. The method showed a linear dynamic range from 0.5 to 50 ng/mL, and the limits of detection and quantification were for both NCOT and COT 0.2 and 0.5 ng/mL, respectively. Validation results were found satisfactory. Finally, the method was applied to the analysis of 60 clinical pediatric samples obtained from Aristotle University's pediatric clinic to check for possible exposure to smoke. Concentration levels ranged between 0.5 and 16.2 ng/mL for NCOT and between 1.0 and 25.1 ng/mL for COT. CONCLUSIONS: A rapid, sensitive, accurate, and simple method was developed and used as a tool for the confirmation of passive smoking in children. It is the first method applied to the analysis of such samples belonging to nonsmokers of young age. The total runtime of the GC-MS analysis was short (20 min), and the pretreatment protocol was simple, giving the ability for analysis of a large number of samples on a daily routine basis.

Determination of Cotinine, 3'-Hydroxycotinine and Nicotine 1'-Oxide in Urine of Passive and Active Young Smokers by LC-Orbitrap-MS/MS Technique.

Tobacco smoke is probably the most significant factor conducing to toxic xenobiotics exposure to humans. The aim of the study was to develop a rapid and sensitive method for the determination of selected nicotine metabolites in urine of tobacco smokers and passive smokers. The method for removing protein and extracting the metabolites involved the centrifugation of urine with acetonitrile. Cotinine, trans-3'-hydroxycotinine, and (2'S)-nicotine 1'-oxide in the supernatant were determined using the LC-Orbitrap-MS/MS technique, with the selected ion monitoring (SIM) and parallel reaction monitoring (PRM) modes used. The recovery of these analytes added to the urine samples ranged from 72% to 101%. Repeatability and reproducibility were less than 3.1% and 10.1%, respectively. The study was carried out among medical students. The group was selected as representatives of young people and who as future physicians should be more aware of the effects of nicotine use. Concentration levels of cotinine and trans-3'-hydroxycotinine determined in ng/mL in the urine of cigarette smokers were 70- and 58-fold higher, respectively, compared to passive smokers. Higher concentrations were recorded in the urine of those passively exposed to tobacco smoke than in non-smokers, confirming that passive exposure to tobacco smoke is not harmless to the human body. However, no significant differences were observed in the concentration of (1'S,2'S)-nicotine 1'-oxide in the samples of individuals from various groups.

[Chronic diarrhoea and weight loss secondary to nicotine use in an adolescent]. / Cas clinique. Diarrhée chronique et perte pondérale suite à la consommation de nicotine chez un adolescent.

Chronic diarrhoea is described as diarrhoea lasting more than 4 weeks. The underlying causes are multiple and the diagnostic orientation depends on several anamnestic and clinical elements. The basic work-up includes biology and stool analysis. We present the case of a 14-year-old adolescent with chronic diarrhoea and weight loss for several months. Extensive complementary analyses were performed, all being totally negative. A careful repeated clinical history revealed that the patient had a significant nicotine intake, confirmed by a urine cotinine test. Withdrawal led to a resolution of the symptomatology. Nicotine consumption in the form of nicotine replacement products among young people is increasing rapidly. Nicotine has multiple health effects, including neurological, gastrointestinal and immune adverse effects.

La diarrhée chronique est décrite comme une diarrhée qui dure plus de 4 semaines. Les étiologies sont multiples et l'orientation diagnostique dépend de plusieurs éléments anamnestiques et cliniques. La mise au point de base comprend une biologie et une analyse de selles. Nous présentons le cas d'un adolescent de 14 ans avec diarrhée chronique et perte pondérale depuis plusieurs mois. Un bilan complémentaire extensif est réalisé et revient totalement négatif. C'est en répétant l'anamnèse qu'une consommation nicotinique non négligeable est mise en évidence chez le patient, confirmée par un dosage élevé de cotinine urinaire. Le sevrage a permis une résolution de la symptomatologie. La consommation de nicotine sous forme de produits de substitution nicotinique chez les jeunes est en forte augmentation. La nicotine a de multiples effets négatifs sur la santé, notamment neurologiques, gastrointestinaux et immunitaires.

Análisis en pelo de nicotina y cotinina para valorar el consumo y la expociión ambiental al tabaco durante el embarazo: un estudio piloto / Hair analysis of nicotine and cotinine for evaluating smoking intake and exposure during pregnancy: a pilot study

En este trabajo se determinó la concentración de nicotina y cotinina en el pelo de un grupo de 19 mujeres embarazadas (9 fumadoras y 10 no fumadoras), en el cuarto mes de embarazo, con la intención de valorar las autodeclaraciones de consumo y exposición ambiental al humo del tabaco durante el embarazo. Las muestras de pelo fueron lavadas con dodecil sulfato sódico al 1 por ciento, digeridas en NaOH y procesadas para el análisis de nicotina y cotinina mediante un radioinmunoensayo tritiado de doble anticuerpo. Las concentraciones de nicotina y cotinina en pelo fueron significativamente diferentes entre los grupos de fumadoras y no fumadoras. Para las mujeres que declararon ser fumadoras activas se obtuvo una concentración media [desviación estandar, DE] de 6,23 [5,17] ng/mg para la nicotina y 0,78 [0,34] ng/mg para la cotinina. Para las mujeres que declararon ser no fumadoras, las concentraciones fueron de 0,91 [1,15] y 0,11 [0,10] ng/mg para la nicotina y cotinina respectivamente. Las concentraciones en pelo de nicotina y cotinina, en las mujeres fumadoras, no correlacionaron con el consumo diario de nicotina, pero se observó correlación (r=0,56) entre el consumo y la concentración urinaria de cotinina. En cambio, las concentraciones en pelo de nicotina y su metabolito sí presentaron correlación (r=0,49 y 0,53 respectivamente9 con la valoración conjunta del consumo diario y la expocición en el mismo grupo de mujeres. En el grupo de las mujeres no-fumadoras, las concentraciones en pelo de nicotina y cotinina se correlacionaron también con la exposición (r=0,93 y 0,78 respectivamente). Además, la concentración de nicotina en pelo pudo discriminar entre varios subgrupos: a) mujeres fumadoras expuestas frente a no-fumadoras no-expuestas, b) mujeres fumadras expuestas frente a no-fuamdoras expuestas, y c) mujeres no-fumadoras expuestas frente a no-fumadoras no-expuestas. Finalmente, la concentración de cotinina pudo discriminar entre mujeres fumadoras y no-fumadoras expuestas así como entre fumadoras y no-fumadoras no-expuestas. Estos resultados confirman el posible uso de la concentración de nicotina y cotinina en pelo como marcador fiable de la expocición sistemática (activa o pasiva) a los constituyentes del humo del tabaco en mujeres embarazadas (AU)