VALIDATION OF ANALYTICAL METHOD OF 2,5-HEXANEDIONE ON URINE BY GAS CHROMATOGRAPHY
Keywords:2,5-hexanedione, Gas chromatography, Hexane, Validation
Objective: The purpose of this study was to obtain a valid analytical method for determining the level of 2,5-hexanedione in the urine of oil industry workers.
Methods: Gas Chromatography (GC) was employed to analyze 2,5-hexanedione in the urine. The analysis was done using HP-5 (Crosslinked methyl siloxane) capillary columns 30 m x 0.320 mm long, film thickness 0.25 μm. The temperature of the detector temperature was 300 °C, and the injector temperature was 250 °C. The helium gas flow rate was 2 ml/min. The detector was Flame Ionization Detection (FID). Parameters of system suitability test and validation were obtained.
Results: This study results that the method of analysis 2,5-hexanedione in urine by Gas Chromatography (GC) confirm the requirements of the validation method with a linearity was 0.99963, accuracy was in the range of 99.16% to 114.13%, the precision with % coefficient of variation was 1.65% to 5.16%, % coefficient variation of specificity was 0.027%, limit of detection was 0.054 μg/ml and limit of quantification was 0.18 μg/ml.
Conclusion: The proposed GC method meets the acceptance criteria of validation parameters and can be applied for routine analysis.
Rusdy MDR. Neurotoxic symptoms analysis due to exposure of xylene organic solvents to paint making workers in company X in 2012. Depok: University of Indonesia; 2012.
Maulida D, Zulkarnaen N. Antioxidant extraction (Lycopene) from tomato fruit using mixed solvents of N-hexane, acetone and ethanol. Semarang: Universtas Diponegoro; 2010.
Cheng X, Wang G, Ma Z, Chen Y, Fan, Jing-Jing, et al. Exposure to 2,5-hexanedione can induce neural malformations in chick embryos. NeuroToxicol 2012;33:1239-47.
Kim Boekelheide. 2,5-Hexanedione alters microtubule assembly. I. Testicular atrophy, not nervous system toxicity, correlates with enhanced tubulin polymerization. Toxicol Appl Pharmacol 1987;88:370-82.
De Caprio AP, O’Neil EA. Alterations in rat axonal cytoskeletal proteins induced by in vitro and in vivo 2,5-hexanedione exposure. Toxicol Pharmacol 1985;78:235.
Couri D, Milks M. Toxicity and metabolism of the neurotoxic hexacarbons n-hexane, 2-hexanone and 2,5-hexanedione. Annu Rev Pharmacol Toxicol 1982;22:145-66.
Cardona A, Marhuenda D, Prieto MJ, Marti J, Periago JF, Sanchez JM, et al. Behaviour of urinary 2,5-hexanedione in occupational co-exposure to n-hexane and acetone. Arch Occup Environ Health 1996;68:88.
American Conference of Governmental Industrial Hygienist (ACGIH). TLVs and BEIs: Based on the documentation of the threshold limit values for chemical substances and physical agents and biological exposure indices. Cincinnati: OH; 2011.
Gori G, Bartolucci GB, Sturaro A, Parvoli G, Doretti L, Troiano R, et al. High-performance liquid chromatographic determination of urinary 2,5-hexanedione as mono-2,4-dinitrophenylhydrazone using ultraviolet detection. J Chromatogr B Biomed Appl 1995;673:165-72.
Oliveira AFF, Patrícia PP, Maria JN, Paiva, Siqueira ME. Determination of 2,5-hexanedione in urine by headspace solid-phase microextraction and gas chromatography. J Anal Toxicol 2009;33:223-8.
Food and Drug Administration, Guidance for Industry: Bioanalytical Method Validation. Maryland: Food and Drug Administration; 2001. Available from: http://www.fda.gov/downloads/DrugsGuidanceComplianceRegulatoryInf ormation/Guidance/UCM070107.pdf. [Last accessed on 11 Sep 2016]
Fedtke N, Bolt HM. Detection of 2,5-hexanedione in the urine of persons not exposed to n-hexane. Int Arch Occup Environ Health 1986;57:143-8.
Harmita. Implementation guidelines for method validation and its calculation. Majalah Ilmu Kefarmasian 2004;1:119-22.
Hasanah AN, Musfirah I, Muchtaridi. Basics of physicochemical analysis in the pharmaceutical field. Yogyakarta: Deepublish; 2016.
The Association of Analytical Communities (AOAC). International Method Committee Guidelines for Validation of Qualitative and Quantitative Food Microbiological Official Methods of Analysis. J AOAC Int 2002;85:1-5.
International Conference on Harmonization, Q2 Validation of analytical procedures : text and methodology international conference on harmonization of technical requirements for registration of pharmaceutical for human use, ICH Harmonized Tripartite Guideline, Canada; 2005.
Maestri L, Ghittori S, Imbriani M, Capodaglio E. Determination of 2,5-hexandione by high-performance liquid chromatography after derivatization with dansyl hydrazine. J Chromatogr B Biomed Appl 1994;657:111-7.