Kokain ve metabolitlerinin insan idrarında analizi
Küçük Resim Yok
Tarih
2016
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Ülkemizde postmortem ve/veya yaşayan canlının biyolojik materyalinde kötüye kullanılabilen maddelerin saptanabilmesine yönelik uygun adli toksikolojik yöntemlerin geliştirilmesi hem tıbbi hem de hukuki açıdan oldukça önemlidir. Çalışmamızda gaz kromatografisi/kütle spektrometresi(GC/MS) kullanılarak kokain ile birlikte onun metabolitleri olan kokaetilen ve benzoilekgoninin insan idrarında analizini gerçekleştirebilmek için kromatografik analiz öncesinde yer alan ekstraksiyon ve türevlendirme basamaklarına yönelik üç farklı metot (Metot A, B ve C) geliştirilmiştir. Metot A'da kromatografik analiz öncesinde türevlendirme uygulanmadan; kokain, kokaetilen ve benzoilekgoninin insan idrarından izole edilebilmesi için hızlı, basit bir sıvı-sıvı ekstraksiyon(LLE) tekniği uygulanmıştır. Uygulanan ekstraksiyon tekniğinde örnekler, petrol eteri ile ekstrakte edilmiştir. Tek aşamalı bir ön hazırlık basamağı sonrası GC/MS ile analiz edilen örneklerde kokain ve kokaetilen saptanabilirken; benzoilekgonin saptanamamıştır. Metot B'de ise Metot A ile aynı olan LLE tekniği örneklere uygulanmış ve ardından örnekler farklı türevlendirme ajanlarıyla (BSTFA+ %1TMCS: N-O-Bis(trimetilsilil)trifloroasetamid+ %1trimetilklorsilan veya PFPA: Pentafloropropiyonik anhidrit) farklı türevlendirme mekanizmaları(sililleme veya açilleme) üzerinden ayrı ayrı türevlendirilmiştir. Ancak GC/MS ile analiz edilen örneklerde benzoilekgonin saptanamamıştır. Metot C'de örnekler, karma modlu katı faz ekstraksiyon(SPE) kartuşu ile ekstrakte edildikten sonra BSTFA:TMCS(99:1) ile türevlendirilmiştir. Örnekler, GC/MS ile analiz edildiğinde benzoilekgonin saptanabilmiştir. LLE ile non-polar ve bazik maddeler olan kokain ve kokaetilen idrardan kolayca izole edilebilirken; polar ve amfoterik madde olan benzoilekgonin izole edilememiştir. Karma modlu SPE kartuş ile farklı polaritede davranış gösteren analitlerin idrardan eş zamanlı ve tek seferde ekstraksiyonunun mümkün olması sebebiyle SPE'nin LLE'ye göre daha avantajlı olduğu düşünülmüştür. Polar yapısı nedeniyle benzoilekgoninin GC/MS analizi öncesinde türevlendirilmesi gerekmektedir. Benzoilekgoninin türevlendirilmesinde BSTFA:TMCS(99:1)'nın, tek başına PFPA'ya kıyasla, daha uygun bir ajan olduğu görülmüştür
In our country, the development of appropriate forensic toxicological methods to detect substances that can be abused in biological material of postmortem and/or living organisms is highly important both in terms of medical and legal aspects. In our study, three different methods(Methods A,B and C) was developed towards the extraction and derivatization steps prior to chromatographic analysis in order to be able to perform the analysis of cocaine and its metabolites cocaethylene and benzoylecgonine in human urine by using gas chromatography/mass spectrometer(GC/MS). A fast, simple liquid-liquid extraction(LLE) technique was performed to isolate cocaine, cocaethylene and benzoylecgonine from human urine, no derivatization was applied before the chromatographic analysis in Method A. Samples were extracted with petroleum ether in the performed extraction technique. After a one-step preliminary step, benzoylecgonine could not be detected while cocaine and cocaethylene could be detected in the samples analyzed by GC/MS. In Method B, the same LLE technique as in Method A was performed to the samples and then the samples were derivatized using the different derivatization mechanisms(silylation or acylation) with the different derivatization agents(BSTFA+1%TMCS:N,O-bis(trimethylsilyl)trifluoroacetamide+1%trimethylchlorosilane or PFPA:Pentafluoropropionic anhydride) separately. However, no benzoylecgonine could be detected in the samples analyzed by GC/MS. In Method C, the samples were derivatized with BSTFA:TMCS(99:1) after being extracted with the mixed mode solid phase extraction(SPE) cartridge. Benzoylecgonine could be detected in the samples analyzed by GC/MS. Polar and amphoteric substance, benzoylecgonine, could not be isolated while non-polar and basic substances, cocaine and cocaethylene, could be easily isolated from urine by LLE. It has been considered that SPE is more advantageous than LLE owing to the fact that it is to be possible one-time and simultaneous extraction of the analytes with different polarity by mixed mode SPE cartridge. It has been necessary to derivatize benzoylecgonine because of its polar structure before GC/MS analysis. BSTFA:TMCS(99:1) has been seen to be a more suitable agent than PFPA alone in the derivatization of benzoylecgonine.
In our country, the development of appropriate forensic toxicological methods to detect substances that can be abused in biological material of postmortem and/or living organisms is highly important both in terms of medical and legal aspects. In our study, three different methods(Methods A,B and C) was developed towards the extraction and derivatization steps prior to chromatographic analysis in order to be able to perform the analysis of cocaine and its metabolites cocaethylene and benzoylecgonine in human urine by using gas chromatography/mass spectrometer(GC/MS). A fast, simple liquid-liquid extraction(LLE) technique was performed to isolate cocaine, cocaethylene and benzoylecgonine from human urine, no derivatization was applied before the chromatographic analysis in Method A. Samples were extracted with petroleum ether in the performed extraction technique. After a one-step preliminary step, benzoylecgonine could not be detected while cocaine and cocaethylene could be detected in the samples analyzed by GC/MS. In Method B, the same LLE technique as in Method A was performed to the samples and then the samples were derivatized using the different derivatization mechanisms(silylation or acylation) with the different derivatization agents(BSTFA+1%TMCS:N,O-bis(trimethylsilyl)trifluoroacetamide+1%trimethylchlorosilane or PFPA:Pentafluoropropionic anhydride) separately. However, no benzoylecgonine could be detected in the samples analyzed by GC/MS. In Method C, the samples were derivatized with BSTFA:TMCS(99:1) after being extracted with the mixed mode solid phase extraction(SPE) cartridge. Benzoylecgonine could be detected in the samples analyzed by GC/MS. Polar and amphoteric substance, benzoylecgonine, could not be isolated while non-polar and basic substances, cocaine and cocaethylene, could be easily isolated from urine by LLE. It has been considered that SPE is more advantageous than LLE owing to the fact that it is to be possible one-time and simultaneous extraction of the analytes with different polarity by mixed mode SPE cartridge. It has been necessary to derivatize benzoylecgonine because of its polar structure before GC/MS analysis. BSTFA:TMCS(99:1) has been seen to be a more suitable agent than PFPA alone in the derivatization of benzoylecgonine.
Açıklama
Anahtar Kelimeler
Adli Tıp, Forensic Medicine, Adli tıp, Forensic medicine, Kokain, Cocaine, Kromatografi-gaz, Chromatography-gas, Kütle spektrometri, Mass spectrometry, Metabolitler, Metabolites, Narkotik bağımlılığı, Narcotic dependence, Narkotikler, Narcotics, Türevlendirme, Derivatization, Özütleme, Extraction, İdrar, Urine
