Kütle Spektrometresi ve Klinik Laboratuvar Uygulamaları
Özet
Kütle spektrometresi, bir numunedeki gaz fazındaki yüklü partiküllerin kütle/yük (m/z) oranlarını ölçerek analitlerin tanımlanması ve miktar tayini için kullanılan "altın standart" bir analitik tekniktir. 19. yüzyılın sonlarında temelleri atılan bu teknoloji, özellikle 1990'larda tandem kütle spektrometrisinin (LC-MS/MS) yenidoğan taramalarında kullanımıyla klinik laboratuvarlarda bir dönüm noktası oluşturmuştur. Sistem temel olarak numune giriş ünitesi, iyonizasyon kaynağı, kütle analizörü, dedektör, vakum kaynağı ve veri sisteminden meydana gelir. Klinik uygulamalarda numune girişi çoğunlukla Gaz Kromatografisi (GC) veya Sıvı Kromatografisi (LC) ile sağlanır. GC-MS uçucu ve küçük moleküller için idealken, LC-MS/MS uçucu olmayan, termal olarak kararsız protein ve hormon gibi kompleks biyomoleküllerin analizine olanak tanır. İyonizasyon aşamasında ESI ve MALDI gibi "yumuşak" teknikler, büyük biyomolekülleri parçalamadan iyonize edebildikleri için modern biyokimyada yaygınlaşmıştır. Kütle analizörleri arasında Quadrupole, TOF ve İyon Tuzakları en sık kullanılan türlerdir. Günümüzde bu teknoloji; toksikoloji, terapötik ilaç izlemi, endokrinoloji, mikrobiyoloji ve metabolik hastalıkların teşhisinde yüksek hassasiyeti ve özgüllüğü nedeniyle vazgeçilmez bir araçtır.
Mass spectrometry is a "gold standard" analytical technique used for the identification and quantification of analytes by measuring the mass-to-charge (m/z) ratios of charged particles in the gas phase generated from a sample. While its foundations were laid in the late 19th century, the technology reached a turning point in clinical laboratories in the 1990s, particularly with the use of tandem mass spectrometry (LC-MS/MS) in newborn screening. The system fundamentally consists of a sample introduction unit, ionization source, mass analyzer, detector, vacuum source, and data system. In clinical applications, sample introduction is predominantly achieved via Gas Chromatography (GC) or Liquid Chromatography (LC). While GC-MS is ideal for volatile and small molecules, LC-MS/MS enables the analysis of complex biomolecules such as proteins and hormones that are non-volatile and thermally unstable. During the ionization stage, "soft" techniques like ESI and MALDI have become widespread in modern biochemistry as they can ionize large biomolecules without fragmentation. Among mass analyzers, Quadrupole, TOF, and Ion Traps are the most frequently used types. Today, this technology is an indispensable tool in toxicology, therapeutic drug monitoring, endocrinology, microbiology, and the diagnosis of metabolic diseases due to its high sensitivity and specificity.
Referanslar
Urban PL. Quantitative mass spectrometry: an overview. Philos Trans A Math Phys Eng Sci. 2016;374(2079):20150382. doi: 10.1098/rsta.2015.0382.
Habib A, Bi L, Hong H, et al. Challenges and strategies of chemical analysis of drugs of abuse and explosives by mass spectrometry. Frontiers in Chemistry Review. 2021;8. doi: 10.3389/fchem.2020.598487.
Brondz I. The newly launched ınternational journal of analytical mass spectrometry and chromatography. Int J Anal Mass Spectrom Chromatogr. 2013;01:1-4. doi: 10.4236/ijamsc.2013.11001.
Griffiths J. A brief history of mass spectrometry. Anal Chem. 2008;80(15):5678-83. doi: 10.1021/ac8013065.
Finehout EJ, Lee KH. An introduction to mass spectrometry applications in biological research. Biochem Mol Biol Educ. 2004;32(2):93-100. doi: 10.1002/bmb.2004.494032020331.
Matern D, Magera MJ. Mass spectrometry methods for metabolic and health assessment. J Nutr. 2001;131(5):1615S-20S. doi: 10.1093/jn/131.5.1615S.
Kaklamanos G, Aprea E, Theodoridis G. (2020). Mass spectrometry: principles and instrumentation. Pico Y (Ed.), Chem Anal Food. (525-552). London: Academic Press.
Rockwood A, Clarke N, Kushnir M. (2017). Mass Spectrometry. Nader Rifai, Carl Wittwer, Rita Horvath (Ed.), Tietz Textbook of Clinical Chemistry and Molecular Diagnostics. (295-323). St. Louis, Missouri: ELSEVIER.
Vorce SP. (2020). Mass Spectrometry. Barry S. Levine, Sarah Kerrigan (Ed.), Principles of Forensic Toxicology. (197-220). Switzerland: Springer AC.
Forgács E, Cserháti T. (2003). Gas chromatography. Lees M (Ed.), Food Authenticity and Traceability. (197-217). Cambridge, England: Woodhead Publishing.
Al-Farga A, Al-Bukhaiti W. Qasim AS, et al. Gas Chromatography: Principles, Advantages and Applications in Food Analysis. Int J Sci Innovs. 2017;6 (1):2319-1473.
Wong YF, Hartmann C, Marriott PJ. Multidimensional gas chromatography methods for bioanalytical research. Bioanalysis. 2014;6(18):2461–2479.
Perez ER, Knapp JA, Horn CK, et al. Comparison of LC-MS-MS and GC-MS analysis of benzodiazepine compounds ıncluded in the drug demand reduction urinalysis program. J Anal Toxicol. 2016;40(3):201-07. doi: 10.1093/jat/bkv140.
Ren J, Zhang A, Kong L, et al. (2021). Aihua Zhang, Wanying Wang (Ed.), Multivariate Data Analysis Approach for Mass Spectrometry‐Based Metabolomics. Mass Spectrometry‐Based Metabolomics in Clinical and Herbal Medicines: Strategies, Technologies and Applications. (45-66). Weinheim, Germany: WILEY-VCH.
Ren J-L, Zhang A-H, Kong L, et al. Advances in mass spectrometry-based metabolomics for investigation of metabolites. RSC Advances. 2018;8:22335-50. doi: 10.1039/C8RA01574K.
Jandera P. (2019). Liquid Chromatography Normal Phase. Worsfold P, Poole C, Townshend A, Miró M (Ed.), Encyclopedia of Analytical Science. (162-73). Oxford, United Kingdom: Academic Press.
Tang DQ, Zou L, Yin XX, et al. HILIC-MS for metabolomics: An attractive and complementary approach to RPLC-MS. Mass Spectrom Rev. 2016;35(5):574-600. doi: 10.1002/mas.21445.
Siuzdak G. An introduction to mass spectrometry ionization: an excerpt from the expanding role of mass spectrometry in biotechnology. J Lab Autom. 2004;9(2):50-63. doi: 10.1016/j.jala.2004.01.004.
Medhe S. Ionization techniques in mass spectrometry: a review. MS&PT. 2018;4. doi: 10.4172/2469-9861.1000126.
Famiglini G, Palma P, Termopoli V, et al. The history of electron ionization in LC-MS, from the early days to modern technologies: A review. Anal Chim Acta, 2021; 1167, 338350. doi:10.1016/j.aca.2021.338350.
Sleeman R, Carter JF. (2005). Mass Spectrometry Overview. Worsfold P, Townshend A, Poole C (Ed.), Encyclopedia of Analytical Science. (337-44). Oxford: Elsevier.
Wilm M. Principles of electrospray ionization. Mol Cell Proteomics. 2011;10(7):M111.009407-M111.07. doi: 10.1074/mcp.M111.009407.
Pitt JJ. Principles and applications of liquid chromatography-mass spectrometry in clinical biochemistry. Clin Biochem Rev. 2009;30(1):19-34.
El-Aneed A, Cohen A, Banoub J. Mass spectrometry, review of the basics: electrospray, MALDI, and commonly used mass analyzers. Appl Spectrosc Rev. 2009;44:210-30. doi: 10.1080/05704920902717872.
Dawson P. Quadrupole mass analyzers: Performance, design and some recent applications. Mass Spectrom Rev. 2005;5:1-37. doi: 10.1002/mas.1280050102.
Dunn WB. (2011). Mass spectrometry in systems biology: an ıntroduction. Jameson D, Verma M, Westerhoff HV (Ed.), Methods Enzymol. (15-35). London, United Kingdom: Academic Press.
Aydoğan C. (2020). Liquid chromatography-high resolution mass spectrometry for the analysis of bioactive natural products. Atta ur R (Ed.), Studies in Natural Products Chemistry. (331-353). Amsterdam, The Netherlands: Elsevier.
van den Ouweland JM, Kema IP. The role of liquid chromatography-tandem mass spectrometry in the clinical laboratory. J Chromatogr B. 2012;883-884:18-32. doi: 10.1016/j.jchromb.2011.11.044.
Fung AWS, Sugumar V, Ren AH, et al. Emerging role of clinical mass spectrometry in pathology. J Clin Pathol. 2020;73(2):61-69. doi: 10.1136/jclinpath-2019-206269.
Gao J, Meyer K, Borucki K, et al. Multiplex Immuno-MALDI-TOF/MS for targeted quantification of protein biomarkers and their proteoforms related to inflammation and renal dysfunction. Anal Chem. 2018; 90. doi: 10.1021/acs.analchem.7b04975.
