Single gas chromatography method with nitrogen phosphorus detector for urinary cotinine determination in passive and active smokers

Nicotine is a major addictive compound in cigarettes and is rapidly and extensively metabolized to several metabolites in humans, including urinary cotinine, considered a biomarker due to its high concentration compared to other metabolites. The aim of this study was to develop a single method for d...

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Main Authors: Lusiane Malafatti, Patrícia Penido Maia, Matheus Coutinho Gonçalves Martins, Maria Elisa Pereira Bastos de Siqueira, Isarita Martins
Format: Article
Language:English
Published: Universidade de São Paulo 2010-12-01
Series:Brazilian Journal of Pharmaceutical Sciences
Subjects:
Online Access:http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-82502010000400019
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author Lusiane Malafatti
Patrícia Penido Maia
Matheus Coutinho Gonçalves Martins
Maria Elisa Pereira Bastos de Siqueira
Isarita Martins
author_facet Lusiane Malafatti
Patrícia Penido Maia
Matheus Coutinho Gonçalves Martins
Maria Elisa Pereira Bastos de Siqueira
Isarita Martins
author_sort Lusiane Malafatti
collection DOAJ
description Nicotine is a major addictive compound in cigarettes and is rapidly and extensively metabolized to several metabolites in humans, including urinary cotinine, considered a biomarker due to its high concentration compared to other metabolites. The aim of this study was to develop a single method for determination of urinary cotinine, in active and passive smokers, by gas chromatography with a nitrogen phosphorus detector (GC-NPD). Urine (5.0 mL) was extracted with 1.0 mL of sodium hydroxide 5 mol L-1, 5.0 mL of chloroform, and lidocaine used as the internal standard. Injection volume was 1 &#956;L in GC-NPD. Limit of quantification was 10 ng mL-1. Linearity was evaluated in the ranges 10-1000 ng mL-1 and 500-6000 ng mL-1, with determination coefficients of 0.9986 and 0.9952, respectively. Intra- and inter-assay standard relative deviations were lower than 14.2 %, while inaccuracy (bias) was less than +11.9%. The efficiency of extraction was greater than 88.5%. Ruggedness was verified, according to Youden's test. Means of cotinine concentrations observed were 2,980 ng mL-1 for active smokers and 132 ng mL-1, for passive smokers. The results revealed that satisfactory chromatographic separation between the analyte and interferents was obtained with a ZB-1 column. This method is reliable, precise, linear and presented ruggedness in the range evaluated. The results suggest that it can be applied in routine analysis for passive and active smokers, since it is able to quantify a wide range of cotinine concentrations in urine.<br>A nicotina é uma substância presente no cigarro capaz de causar dependência, sendo biotransformada em vários metabólitos nos seres humanos, dentre eles a cotinina urinária, que é considerada um indicador biológico de exposição à nicotina, devido a suas altas concentrações, comparado a outras matrizes. Assim, o objetivo deste estudo foi desenvolver um único método para determinação de cotinina urinária, em amostras de urina de fumantes ativos e passivos, através de cromatografia em fase gasosa com detector de nitrogênio- fósforo (CG-DNF). Para o preparo de amostras foram utilizados 5 mL de urina, 1 mL de hidróxido de sódio 5 mol L-1, 5 mL de clorofórmio, tendo como padrão interno a lidocaína. Na faixa de concentrações de 10-1000 ng mL-1 e 500- 6000 ng mL-1, o coeficiente de determinação foi 0,9986 e 0,9952, respectivamente e, o limite de quantificação foi 10 ng mL-1. A precisão intra- e interensaio apresentou desvio padrão relativo (%) menor que 14,2% e a inexatidão foi menor que +11,9%, com uma eficiência de extração de 88,5%. O método apresentou robustez, de acordo com o teste de Youden. As concentrações médias de cotinina observadas foram 2980 ng mL-1, para fumantes ativos e 132 ng mL-1, para fumantes passivos. Os resultados sugerem que o método é confiável, preciso, linear e apresentou robustez, na faixa avaliada, podendo ser aplicado na rotina para análises de amostras de fumantes ativos e passivos, pois é capaz de quantificar uma ampla faixa de concentrações de cotinina urinária.
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spelling doaj.art-35acf56b754046a18427bda8088b1ba02022-12-22T03:13:02ZengUniversidade de São PauloBrazilian Journal of Pharmaceutical Sciences1984-82502175-97902010-12-0146476977610.1590/S1984-82502010000400019Single gas chromatography method with nitrogen phosphorus detector for urinary cotinine determination in passive and active smokersLusiane MalafattiPatrícia Penido MaiaMatheus Coutinho Gonçalves MartinsMaria Elisa Pereira Bastos de SiqueiraIsarita MartinsNicotine is a major addictive compound in cigarettes and is rapidly and extensively metabolized to several metabolites in humans, including urinary cotinine, considered a biomarker due to its high concentration compared to other metabolites. The aim of this study was to develop a single method for determination of urinary cotinine, in active and passive smokers, by gas chromatography with a nitrogen phosphorus detector (GC-NPD). Urine (5.0 mL) was extracted with 1.0 mL of sodium hydroxide 5 mol L-1, 5.0 mL of chloroform, and lidocaine used as the internal standard. Injection volume was 1 &#956;L in GC-NPD. Limit of quantification was 10 ng mL-1. Linearity was evaluated in the ranges 10-1000 ng mL-1 and 500-6000 ng mL-1, with determination coefficients of 0.9986 and 0.9952, respectively. Intra- and inter-assay standard relative deviations were lower than 14.2 %, while inaccuracy (bias) was less than +11.9%. The efficiency of extraction was greater than 88.5%. Ruggedness was verified, according to Youden's test. Means of cotinine concentrations observed were 2,980 ng mL-1 for active smokers and 132 ng mL-1, for passive smokers. The results revealed that satisfactory chromatographic separation between the analyte and interferents was obtained with a ZB-1 column. This method is reliable, precise, linear and presented ruggedness in the range evaluated. The results suggest that it can be applied in routine analysis for passive and active smokers, since it is able to quantify a wide range of cotinine concentrations in urine.<br>A nicotina é uma substância presente no cigarro capaz de causar dependência, sendo biotransformada em vários metabólitos nos seres humanos, dentre eles a cotinina urinária, que é considerada um indicador biológico de exposição à nicotina, devido a suas altas concentrações, comparado a outras matrizes. Assim, o objetivo deste estudo foi desenvolver um único método para determinação de cotinina urinária, em amostras de urina de fumantes ativos e passivos, através de cromatografia em fase gasosa com detector de nitrogênio- fósforo (CG-DNF). Para o preparo de amostras foram utilizados 5 mL de urina, 1 mL de hidróxido de sódio 5 mol L-1, 5 mL de clorofórmio, tendo como padrão interno a lidocaína. Na faixa de concentrações de 10-1000 ng mL-1 e 500- 6000 ng mL-1, o coeficiente de determinação foi 0,9986 e 0,9952, respectivamente e, o limite de quantificação foi 10 ng mL-1. A precisão intra- e interensaio apresentou desvio padrão relativo (%) menor que 14,2% e a inexatidão foi menor que +11,9%, com uma eficiência de extração de 88,5%. O método apresentou robustez, de acordo com o teste de Youden. As concentrações médias de cotinina observadas foram 2980 ng mL-1, para fumantes ativos e 132 ng mL-1, para fumantes passivos. Os resultados sugerem que o método é confiável, preciso, linear e apresentou robustez, na faixa avaliada, podendo ser aplicado na rotina para análises de amostras de fumantes ativos e passivos, pois é capaz de quantificar uma ampla faixa de concentrações de cotinina urinária.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-82502010000400019Cromatografia em fase gasosaCromatografia em fase gasosaCotinina urináriaUrinaFumantesGas chromatographyUrinary cotinineUrineSmokers
spellingShingle Lusiane Malafatti
Patrícia Penido Maia
Matheus Coutinho Gonçalves Martins
Maria Elisa Pereira Bastos de Siqueira
Isarita Martins
Single gas chromatography method with nitrogen phosphorus detector for urinary cotinine determination in passive and active smokers
Brazilian Journal of Pharmaceutical Sciences
Cromatografia em fase gasosa
Cromatografia em fase gasosa
Cotinina urinária
Urina
Fumantes
Gas chromatography
Urinary cotinine
Urine
Smokers
title Single gas chromatography method with nitrogen phosphorus detector for urinary cotinine determination in passive and active smokers
title_full Single gas chromatography method with nitrogen phosphorus detector for urinary cotinine determination in passive and active smokers
title_fullStr Single gas chromatography method with nitrogen phosphorus detector for urinary cotinine determination in passive and active smokers
title_full_unstemmed Single gas chromatography method with nitrogen phosphorus detector for urinary cotinine determination in passive and active smokers
title_short Single gas chromatography method with nitrogen phosphorus detector for urinary cotinine determination in passive and active smokers
title_sort single gas chromatography method with nitrogen phosphorus detector for urinary cotinine determination in passive and active smokers
topic Cromatografia em fase gasosa
Cromatografia em fase gasosa
Cotinina urinária
Urina
Fumantes
Gas chromatography
Urinary cotinine
Urine
Smokers
url http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-82502010000400019
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