HPTLC assay of nicotine and cotinine in biological samples
This study presents the development of a simple high-performance thin layer chromatography (HPTLC) method for the determination of nicotine and its metabolite cotinine in human plasma and urine. The following mobile phases: methanol: ammonia (100:1.5, v:v), chloroform: acetone: ammonia (48.75: 48.7...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Ion Motofei, Carol Davila University
2018-10-01
|
| Series: | Journal of Mind and Medical Sciences |
| Subjects: | |
| Online Access: | https://scholar.valpo.edu/cgi/viewcontent.cgi?article=1146&context=jmms |
| _version_ | 1818082723025125376 |
|---|---|
| author | Ana Maria Vlasceanu Daniela Luiza Baconi Anne Marie Ciobanu Vlad D. Constantin Liana Pleș Cristian Balalau Miriana Stan |
| author_facet | Ana Maria Vlasceanu Daniela Luiza Baconi Anne Marie Ciobanu Vlad D. Constantin Liana Pleș Cristian Balalau Miriana Stan |
| author_sort | Ana Maria Vlasceanu |
| collection | DOAJ |
| description | This study presents the development of a simple high-performance thin layer chromatography (HPTLC) method for the determination of nicotine and its metabolite cotinine in human plasma and urine.
The following mobile phases: methanol: ammonia (100:1.5, v:v), chloroform: acetone: ammonia (48.75: 48.75: 2.5, v:v:v), methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) and glass plates precoated with silicagel 60 F254 (20x20) as a stationary phase were used. Densitometric scanning was performed at 263 nm. Two different extraction procedures have been applied: liquid-liquid extraction using dichloromethane at alkaline pH and solid-phase extraction using C18 cartridges.
Preliminary tests in order to establish the system of solvents for development, as well as the range of linearity, were conducted. The best separation of nicotine and cotinine was obtained by using methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) as the mobile phase. The liquid-liquid extraction technique led to better results than solid phase extraction. The regression curves were linear (with a corresponding correlation coefficient higher than 0.99) in the quantities range of 200 ng–1000 ng/spot for both nicotine and cotinine. The UV spectra confirm the identification of nicotine and cotinine both in the standards and in the extracts after liquid-liquid extraction.
The proposed method can be applied for the simultaneous evaluation of nicotine and cotinine in biological samples at toxic/lethal levels. Thus, the method may be applicable in lethal nicotine intoxication cases in forensic toxicological analysis. |
| first_indexed | 2024-12-10T19:26:38Z |
| format | Article |
| id | doaj.art-5301d6f9cc854fc3a9f11f16424d11eb |
| institution | Directory Open Access Journal |
| issn | 2392-7674 2392-7674 |
| language | English |
| last_indexed | 2024-12-10T19:26:38Z |
| publishDate | 2018-10-01 |
| publisher | Ion Motofei, Carol Davila University |
| record_format | Article |
| series | Journal of Mind and Medical Sciences |
| spelling | doaj.art-5301d6f9cc854fc3a9f11f16424d11eb2022-12-22T01:36:21ZengIon Motofei, Carol Davila UniversityJournal of Mind and Medical Sciences2392-76742392-76742018-10-015224424910.22543/7674.52.P244249HPTLC assay of nicotine and cotinine in biological samplesAna Maria Vlasceanu0Daniela Luiza Baconi1Anne Marie Ciobanu2Vlad D. Constantin3Liana Pleș4Cristian Balalau5Miriana Stan6Carol Davila University of Medicine and Pharmacy, Faculty of Pharmacy, Bucharest, RomaniaCarol Davila University of Medicine and Pharmacy, Faculty of Pharmacy, Bucharest, RomaniaCarol Davila University of Medicine and Pharmacy, Faculty of Pharmacy, Bucharest, RomaniaCarol Davila University of Medicine and Pharmacy, Faculty of General Medicine, Bucharest, RomaniaCarol Davila University of Medicine and Pharmacy, Faculty of General Medicine, Bucharest, RomaniaCarol Davila University of Medicine and Pharmacy, Faculty of General Medicine, Bucharest, RomaniaCarol Davila University of Medicine and Pharmacy, Faculty of Pharmacy, Bucharest, RomaniaThis study presents the development of a simple high-performance thin layer chromatography (HPTLC) method for the determination of nicotine and its metabolite cotinine in human plasma and urine. The following mobile phases: methanol: ammonia (100:1.5, v:v), chloroform: acetone: ammonia (48.75: 48.75: 2.5, v:v:v), methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) and glass plates precoated with silicagel 60 F254 (20x20) as a stationary phase were used. Densitometric scanning was performed at 263 nm. Two different extraction procedures have been applied: liquid-liquid extraction using dichloromethane at alkaline pH and solid-phase extraction using C18 cartridges. Preliminary tests in order to establish the system of solvents for development, as well as the range of linearity, were conducted. The best separation of nicotine and cotinine was obtained by using methanol: chloroform: ammonia (48.75: 48.75: 0.5, v:v:v) as the mobile phase. The liquid-liquid extraction technique led to better results than solid phase extraction. The regression curves were linear (with a corresponding correlation coefficient higher than 0.99) in the quantities range of 200 ng–1000 ng/spot for both nicotine and cotinine. The UV spectra confirm the identification of nicotine and cotinine both in the standards and in the extracts after liquid-liquid extraction. The proposed method can be applied for the simultaneous evaluation of nicotine and cotinine in biological samples at toxic/lethal levels. Thus, the method may be applicable in lethal nicotine intoxication cases in forensic toxicological analysis.https://scholar.valpo.edu/cgi/viewcontent.cgi?article=1146&context=jmmsnicotinecotinineTLC-densitometryliquid-liquid extraction |
| spellingShingle | Ana Maria Vlasceanu Daniela Luiza Baconi Anne Marie Ciobanu Vlad D. Constantin Liana Pleș Cristian Balalau Miriana Stan HPTLC assay of nicotine and cotinine in biological samples Journal of Mind and Medical Sciences nicotine cotinine TLC-densitometry liquid-liquid extraction |
| title | HPTLC assay of nicotine and cotinine in biological samples |
| title_full | HPTLC assay of nicotine and cotinine in biological samples |
| title_fullStr | HPTLC assay of nicotine and cotinine in biological samples |
| title_full_unstemmed | HPTLC assay of nicotine and cotinine in biological samples |
| title_short | HPTLC assay of nicotine and cotinine in biological samples |
| title_sort | hptlc assay of nicotine and cotinine in biological samples |
| topic | nicotine cotinine TLC-densitometry liquid-liquid extraction |
| url | https://scholar.valpo.edu/cgi/viewcontent.cgi?article=1146&context=jmms |
| work_keys_str_mv | AT anamariavlasceanu hptlcassayofnicotineandcotinineinbiologicalsamples AT danielaluizabaconi hptlcassayofnicotineandcotinineinbiologicalsamples AT annemarieciobanu hptlcassayofnicotineandcotinineinbiologicalsamples AT vladdconstantin hptlcassayofnicotineandcotinineinbiologicalsamples AT lianaples hptlcassayofnicotineandcotinineinbiologicalsamples AT cristianbalalau hptlcassayofnicotineandcotinineinbiologicalsamples AT mirianastan hptlcassayofnicotineandcotinineinbiologicalsamples |