Amfetamin tipi stimülanların idrarda analizine yönelik katı faz mikroekstraksiyon yöntemi geliştirilmesi
Küçük Resim Yok
Dosyalar
Tarih
2021
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/embargoedAccess
Özet
Madde kullanımı dünyada olduğu gibi ülkemizde de gittikçe artmaktadır ve tüm boyutlarıyla ortaya konması gereken bir konudur. Amfetamin Tipi Stimülanlar (ATS) dünya genelinde esrardan sonra en çok suistimal edilen güçlü merkezi sinir sistemi uyarıcılarıdır. Bu maddeleri kullananların saptanması, dolayısıyla toksikolojik analizleri madde kullanımı ile mücadelede büyük bir yer tutmaktadır. ATS'lerin analizinde sıklıkla kullanılan idrar ve kan gibi biyolojik materyallerde ilgili analiti saptayabilmek için hızlı, temiz ve uygun ön işlem (ekstraksiyon) prosedürlerinin uygulanması önemlidir. Günümüze kadar birçok analitik yöntem geliştirilmiş olmakla birlikte daha az çözgenin harcandığı, yeşil çevreye katkıda bulunan ve küçük örnek hacimlerinde çalışma imkânı sağlayan mikroekstraksiyon yöntemleri üzerine yoğunlaşılmaya başlanmıştır. 1990'ların başında geleneksel örnek hazırlamadaki bazı dezavantajları gidermek, yerinde (on-site) analize olanak sağlamak vb. amaçlarla katı faz mikroekstraksiyon yöntemi (SPME) analitik uygulamaya sokulmuştur. Fiber tip SPME tekniği, günümüze kadar en çok kullanılan SPME yaklaşımı olmaya devam etmektedir. Uygun fiber yardımıyla çözücü kullanılmadan yapılan bu teknik; sıvıdan bileşenleri ya tepe boşluğundan (headspace) ya da sıvı içerisine daldırarak doğrudan ekstrakte edilmesi prensibine dayanmaktadır. Bu tez çalışmasında, kromatografik ayırma tekniklerinden yararlanılarak ATS'lerden amfetamin (AMP), metamfetamin (MET), 3,4- metilendioksiamfetamin (MDA), 3,4-metilendioksimetamfetamin (MDMA,ekstazi)'nin idrarda hassas ve güvenilir tayini için SPME yöntemi geliştirilmesi amaçlanmıştır. Ticari olarak satılan fiberlerden uygun olan Polidimetilsiloksan/Divinilbenzen (PDMS/DVB) fiberi ile yöntem geliştirilmiştir. Geliştirilen yöntem kemometrik olarak optimize edildikten sonra valide edilmiştir. Nanomalzemeler kullanılarak elektrokimyasal polimerizasyonla kaplama yapılarak laboratuvar tipi fiberler oluşturulmuştur. Oluşturulan bu fiberler kullanılarak ticari olarak alınan fiberle valide edilen yöntem ATS'lerin idrarda analizinde uygulanmaya çalışılmıştır. Geliştirilen üst boşluklu (head space) SPME yönteminde hiçbir çözgen kullanılmamış olup, yeşil çevreye büyük katkı sağlayan bir yöntem oluşturulmuştur. Ancak, çalışma kapsamında yöntem geliştirme kısmında yaşanılan en büyük sıkıntı fiberin türevlendirici nedeniyle soyulması olmuş ve fiber çok az tekrarla kullanılabilmiştir. Literatürlerde genelde kullanılan türevlendirici reaktiflerden AA, PFPA ve TFA'nın etkinlikleri karşılaştırılmış ve TFA ile daha güzel sonuçlar elde edilmiştir. Plakett-Burman tasarımı sonrası regresyon analizinde adsorpsiyon süresi, türevlendirme sıcaklığı ve türevlendirme süresinin en etkili faktörler olduğu belirlenmiştir. Bu faktörlerin optimize edilmesiyle en iyi sonuçlar 10 dk. adsorpsiyon süresinde, 800C türevlendirme sıcaklığında ve 1 dk. türevlendirme süresinde elde edilmiştir. Kemometrik olarak optimize edilen yöntem; seçimlilik, kesinlik, doğruluk, belirtme alt sınırı (LOD), saptama alt sınırı (LOQ), doğrusallık, duyarlık, çalışma aralığı, taşınma etkisi (carry over) ve matriks etkisi parametreleri yönünden valide edilmiştir. Nanoteknolojik gelişmeler sayesinde ayırma bilimi alanında maddelerin eser analizi için yeni sorbentler olarak karbon nanotüpler (CNT) kullanılmaktadır. Bu çalışmada, laboratuvar tipi fiber elde edebilmek için monomer olarak pirol varlığında MWCNT, MWCNT-COOH, azot-katkılı MWCNT, grafen ve nanoselüloz ayrı ayrı elektropolimerizasyon yöntemiyle çelik tele kaplanmıştır. Ancak, geliştirilen SPME yöntemi, laboratuvar tipi fiberlere uygulandığında ATS'leri ait bir analit piki saptanamamıştır. Kısıtlı imkanlar nedeniyle nanomalzemelerle yapılan çalışmalar daha ileri aşamalara götürülememiştir. Bu doktora tezi ile ATS'lerin analizi için çevreye duyarlı ve hiç çözgenin kullanılmadığı, kısa sürede eş zamanlı ekstraksiyona olanak sağlayan SPME yöntemi geliştirilmiştir. Türevlendirme basamağının fiberlere verdiği zarar somut olarak gösterilmiştir. SPME yöntemi birçok alanda kullanılmasına rağmen adli toksikoloji alanında kullanımı kısıtlıdır. Bu çalışma ile geliştirilen yöntem rutin analize olanak sağlamaktadır. Özellikle otomatize edilebilirliğiyle kullanım alanı kısıtlılıktan çıkıp, genişleyebilmektedir.
Substance use is increasing in our country as well as in the world, and it is an issue that needs to be addressed in all its dimensions. Amphetamine-type stimulants (ATS) are the most abused potent central nervous system stimulants worldwide after cannabis. Identification substance users, and therefore their toxicological analysis, has a great place in the anti drug effort. It is important to apply fast, clean and appropriate pretreatment (extraction) procedures in order to detect the relevant analyte in biological materials such as urine and blood, which are frequently used in the analysis of ATSs. Although many analytical methods have been developed to date, it has begun to focus on microextraction methods that use less solvent, contribute to a green environment and allow working in small sample volumes. In the early 1990s, it was aimed to overcome some of the disadvantages of traditional sample preparation, to allow on-site analysis, etc. for these purposes, the solid phase microextraction method (SPME) has been put into analytical application. The fiber type SPME technique remains the most widely used SPME approach to date. This technique is made without the use of solvents with the help of suitable fiber and based on the principle of directly extracting the components from the liquid either from the headspace or by immersing them in the liquid. In this thesis, SPME method is developed for the sensitive and reliable determination of amphetamine (AMP), methamphetamine (MET), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) from ATSs in urine by using chromatographic separation techniques. The method was developed with suitable Polydimethylsiloxane/Divinylbenzene (PDMS/DVB) fiber from commercially available fibers. The developed method was validated after being optimized chemometrically. Laboratory type fibers were formed by coating with electrochemical polymerization using nanomaterials. Using these fibers, the method validated with commercially available fiber has been tried to be applied in the analysis of ATSs in urine. No solvent was used in the developed head space SPME method, and a method that greatly contributed to the green environment was created. However, the biggest problem experienced in the method development part of the study was the peeling of the fiber due to the derivatization and the fiber could be used with very little repetition. The efficiency of AA, PFPA and TFA, which are generally used in the literature, were compared and better results were obtained with TFA. It was determined that adsorption time, derivatization temperature and derivatization time were the most effective factors in the regression analysis after Plackett-Burman design. The best results were obtained in 10 min. adsorption time, 800C derivatization temperature and 1 min. derivatization time. Chemometrically optimized method was validated for selectivity, precision, accuracy, limit of detection (LOD), limit of quantitation (LOQ), linearity, sensitivity, working range, carry over, and matrix effect parameters. Thanks to nanotechnological developments, carbon nanotubes (CNT) are used as new sorbents for trace analysis of substances in the field of separation science. In this study, in the presence of pyrrole as monomer, MWCNT, MWCNT-COOH, nitrogen-doped MWCNT, graphene and nanocellulose were coated separately on steel wire by electropolymerization method in order to obtain laboratory type fiber. However, when the developed SPME method was applied to laboratory type fibers, no analyte peak of ATS could be detected. Due to limited opportunities, studies with nanomaterials could not be taken to further stages. With this doctoral thesis, the SPME method was developed for the analysis of ATSs, which was environmentally friendly, did not use any solvent, and allowed simultaneous extraction in a short time. The damage done by the derivatization step to the fibers was concretely demonstrated. Although the SPME method is used in many fields, its use in forensic toxicology is limited. The method developed in this study allows routine analysis. Especially with its automatizability, the usage area can get out of the limitation and expand.
Substance use is increasing in our country as well as in the world, and it is an issue that needs to be addressed in all its dimensions. Amphetamine-type stimulants (ATS) are the most abused potent central nervous system stimulants worldwide after cannabis. Identification substance users, and therefore their toxicological analysis, has a great place in the anti drug effort. It is important to apply fast, clean and appropriate pretreatment (extraction) procedures in order to detect the relevant analyte in biological materials such as urine and blood, which are frequently used in the analysis of ATSs. Although many analytical methods have been developed to date, it has begun to focus on microextraction methods that use less solvent, contribute to a green environment and allow working in small sample volumes. In the early 1990s, it was aimed to overcome some of the disadvantages of traditional sample preparation, to allow on-site analysis, etc. for these purposes, the solid phase microextraction method (SPME) has been put into analytical application. The fiber type SPME technique remains the most widely used SPME approach to date. This technique is made without the use of solvents with the help of suitable fiber and based on the principle of directly extracting the components from the liquid either from the headspace or by immersing them in the liquid. In this thesis, SPME method is developed for the sensitive and reliable determination of amphetamine (AMP), methamphetamine (MET), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) from ATSs in urine by using chromatographic separation techniques. The method was developed with suitable Polydimethylsiloxane/Divinylbenzene (PDMS/DVB) fiber from commercially available fibers. The developed method was validated after being optimized chemometrically. Laboratory type fibers were formed by coating with electrochemical polymerization using nanomaterials. Using these fibers, the method validated with commercially available fiber has been tried to be applied in the analysis of ATSs in urine. No solvent was used in the developed head space SPME method, and a method that greatly contributed to the green environment was created. However, the biggest problem experienced in the method development part of the study was the peeling of the fiber due to the derivatization and the fiber could be used with very little repetition. The efficiency of AA, PFPA and TFA, which are generally used in the literature, were compared and better results were obtained with TFA. It was determined that adsorption time, derivatization temperature and derivatization time were the most effective factors in the regression analysis after Plackett-Burman design. The best results were obtained in 10 min. adsorption time, 800C derivatization temperature and 1 min. derivatization time. Chemometrically optimized method was validated for selectivity, precision, accuracy, limit of detection (LOD), limit of quantitation (LOQ), linearity, sensitivity, working range, carry over, and matrix effect parameters. Thanks to nanotechnological developments, carbon nanotubes (CNT) are used as new sorbents for trace analysis of substances in the field of separation science. In this study, in the presence of pyrrole as monomer, MWCNT, MWCNT-COOH, nitrogen-doped MWCNT, graphene and nanocellulose were coated separately on steel wire by electropolymerization method in order to obtain laboratory type fiber. However, when the developed SPME method was applied to laboratory type fibers, no analyte peak of ATS could be detected. Due to limited opportunities, studies with nanomaterials could not be taken to further stages. With this doctoral thesis, the SPME method was developed for the analysis of ATSs, which was environmentally friendly, did not use any solvent, and allowed simultaneous extraction in a short time. The damage done by the derivatization step to the fibers was concretely demonstrated. Although the SPME method is used in many fields, its use in forensic toxicology is limited. The method developed in this study allows routine analysis. Especially with its automatizability, the usage area can get out of the limitation and expand.
