JASMI  Vol.2 No.2 , June 2012
Organic Solvent-Free and Simple Method for Determining Cyromazine and Its Metabolite, Melamine, in Cow’s Milk
Abstract: This paper described an organic solvent-free, rapid, simple, and space-saving method of sample preparation followed by HPLC coupled photo-diode array (PDA) detector for simultaneous quantification of cyromazine (CYR) and its decy-cropropylated metabolite, melamine (MEL), in milk. The HPLC-PDA was performed on an Inertsil? HILIC column with an isocratic aqueous mobile phase. Analytes were extracted from the sample using water, and purified by Mono-Spin?-C18, a centrifugal monolithic silica spin mini-columns, and quantified within 20 min. The method, performed under 100% aqueous conditions, obtained average recoveries for CYR and MEL in the range of 93.2% - 99.1% with relative standard deviations ≤ 2.8%. The quantitation limits were 8.5 ng/mL for CYR and 10 ng/mL for MEL, respectively. No organic solvents were used at any stage of the analysis.
Cite this paper: N. Furusawa, "Organic Solvent-Free and Simple Method for Determining Cyromazine and Its Metabolite, Melamine, in Cow’s Milk," Journal of Analytical Sciences, Methods and Instrumentation, Vol. 2 No. 2, 2012, pp. 68-73. doi: 10.4236/jasmi.2012.22013.

[1]   FAO, Pesticide Management, JMPR Evaluation, List of Pesticides Evaluated by JMPR and JMPS, Cyromazine, JMPR No. 169, 2009 (R) Evaluation.

[2]   A. K. Hau, T. H. Kwan and P. K. Li, “Melamine Toxicity and the Kidney,” Journal of the American Society Nephrology, Vol. 20, 2009, pp. 245-250. doi:10.1681/ASN.2008101065

[3]   MSDS,

[4]   International Chemical Safety Card,

[5]   OSHA, “Chemical Sampling Information,”

[6]   WHO, “Some Chemicals That Care Tumors of the Kidney or Urinary Bladder in Rodents and Some Other Substances,”

[7]   H. D. Heck and R. W. Tyl, “The Induction of Bladder Stones by Terephthalic Acid, Dimethyl Terephthalate, and Melamine (2,4,6-Triamino-s-triazine) and Its Relevance to Risk Assessment,” Regulatory Toxicology and Pharmacology, Vol. 5, No. 3, 1985, pp. 294-313. doi:10.1016/0273-2300(85)90044-3

[8]   FDA, “Protecting and Promoting Your Health,” 164520.htm

[9]   WHO, Media Centre, International Experts Limit Melamine Levels in Food,

[10]   Codex, Resticide Residues in Food and Feed, Cyromazine,

[11]   USDA FSIS, “Cyromazine and Melamine,” July 1991, Retrieved 27 April 2007.

[12]   S. Tittlemier, Health Canada, “Background Paper on Methods for the Analysis of Melamine and Related Compounds in Foods and Animal Feeds,” Ottawa, 1-4 December 2008.

[13]   H. A. Yu, Y. F. Tao, D. M. Chen, Y. L. Wang, Z. Y. Liu, Y. H. Pan, L. L. Huang, D. P. Peng, M. H. Dal, Z. L. Liu and Z. H. Yuan, “Development of a High Performance Liquid Chromatography Method and Liquid Chromatography-Tandem Mass Spectrometry Method with Pressurized Liquid Extraction for Simultaneous Quantification and Confirmation of Cyromazine, Melamine and Its Metabolites in Foods of Animal Origin,” Analytica Chimica Acta, Vol. 682, No. 1-2, 2010, pp. 48-58. doi:10.1016/j.aca.2010.09.032

[14]   S. S. Wang, D. M. Li, Z. D. Hua and M. P. Zhao, “Molecularly Imprinted Monolith Coupled On-Line with High Performance Liquid Chromatography for Simultaneous Quantitative Determination of Cyromazine and Melamine,” Analyst, Vol. 136, No. 18, 2011, pp. 3672-3679. doi:10.1039/c1an15086c

[15]   H. W. Sun, L. X. Wang, N. Liu, F. X. Qiao and S. X. Liang, “SPE Then RP-LC for Simultaneous Analysis of Cyromazine and Its Metabolite Melamine in Liquid Milk and Egg,” Chromatographia, Vol. 70, No. 11, 2009, pp. 1685-1689. doi:10.1365/s10337-009-1355-x

[16]   R. C. Wei, R. Wang, Q. F. Zeng, M. Chen and T. Z. Lui, “High-Performance liquid Chromatographic Method for the Determination of cyromazine and Melamine Residues in Milk and Pork,” Journal of Chromatographic Science, Vol. 47, 2009, pp. 581-584.

[17]   P. T. Anastas, J. C. Warner, “Green Chemistry: Theory and Practice,” Oxford University Press, Oxford, 1998.

[18]   T. Yoshimura, T. Nishinomiya, Y. Honda and M. Murabayashi, “Green Chemistry—Aim for the Zero Emission-Chemicals,” Sankyo Publishing Co. Ltd. Press, Tokyo, Japan, 2001.

[19]   L. Huber (Ed.), “Validation and Quantification in Analytical Laboratories,” Interpharm Press, East England, 1998.

[20]   ICH, “Work Products, ICH Guidelines, Quality Guidelines,”

[21]   AOAC International, “Method Validation, Guidelines & References,”

[22]   FDA/CDER/CVM, “Guidelines for Industry—Bioanalytical Method Validation,” 2001.

[23]   Codex Alimentarius Commission, ALINORM 01/23, Codex Committee on Methods of Analysis and Sampling, Joint FAO/WHO Food Standards Programme, the 23rd Section of the Codex Committee on Method of Analysis and Sampling.

[24]   GL Sciences, Monolithic SPE Column for the Purification and Enrichment of Small Amount Sample, MonoSpin? Series.