MALDI-TOF 1, # Quantitative Analysis of Cashmere and Other Animal Hair Fibers in Textiles Using MALDI-TOF Mass Spectrometry Shinichi Ohashi 1,#,
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1 MALDI-TOF 1, # Quantitative Analysis of Cashmere and Other Animal Hair Fibers in Textiles Using MALDI-TOF Mass Spectrometry Shinichi Ohashi 1,#, Yuka Demura 1, Motoaki Sano 1, and Yoichiro Yoshioka 2 1 Genome Biotechnology Laboratory, Kanazawa Institute of Technology, 3-1 Yatsukaho, Hakusan, Ishikawa , Japan 2 Boken Quality Evaluation Institute, Ue-Machi, Chuo-Ku, Osaka , Japan Abstract : Techniques for analyzing the composition of animal hair fibers in textiles are very important to guarantee the quality of animal hair yarns and fabrics. We have recently developed new methods based on sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption/ionization-time of flight (MALDI- TOF) mass spectroscopy in order to identify the animal species from the hair fibers. In this study, we assigned the animal species-specific peaks by tandem mass (MS/MS) analysis. We prepared cashmere-wool and cashmere-yak standard samples and observed very good linearity between the composition and the peak area ratio. The quantitative method was validated using nine blind-test samples containing known contents of cashmere, wool, and yak. The method was also validated by measuring various samples containing cashmere and other animal hair fibers, natural fibers and chemical fibers and by comparing the results with those obtained by optical microscopy. (Received 14 February, 2014 ; Accepted 6 March, 2014) # corresponding author 1 2 DNA DNA 2 DNA DNA 5 I SDS-PAGE MALDI- TOF m/e
2 MALDI-TOF CBB MALDI-TOF xcise Proteome Systems CHCA MALDI-TOF AXIMA-CFRplus Matrix Science Mascot NCBI MSDB CCMI 12 DTT SDS IAA Bio-Rad 6 SDS-PAGE nano-lc KY CHCA MALDI TOF/TOF 4800 plus AB Sciex MS/MS NCBI Sigma-Aldrich HPLC HPLC Promega α- -4- CHCA Sigma-Aldrich MM mg 1.5 ml 50 mmol/l DTT 4% SDS 0.1 M ph ml rpm mmol/l IAA mmol/l DTT10μl rpm 5 DTT SDS SDS-PAGE e 90 mm 83 mm 1mm AE-6513P CBB SDS-PAGE SDS DTT SDS-PAGE MALDI-TOF m/e MALDI-TOF MS/MS 6 SDS-PAGE LC MALDI-TOF MS/MS Table 1 m/e= A
3 m/e= B m/e=2504 keratin type I microfibrillar 47.6 kdalike A 33B N- NCBI BLAST 33A Table 2 keratin33a m/e= hair acidic keratin 1 keratin type I cuticular Ha 4-like Table 1 MS/ MS Table 1 Results of MS/MS analysis of animal-specific peaks. Table 2 Amino acid sequence of animal species-specific region in keratin type I.
4 MALDI-TOF MALDI 8,9 6 SDS-PAGE I MALDI-TOF 80% 50% 20% Fig. 1 Cashmere Peak % PAC 100/ PAc+PAw Cashmere Peak % PAC 100/ PAc+PAY PAC : PAW : PAY : Fig. 2 Calibration curve for cashmere/wool mixture. Fig. 3 Calibration curve for cashmere/yak mixture. Fig. 1 Mass spectra of cashmere/wool mixture. Fig. 2 Fig SDS-PAGE
5 Fig. 4 Fig. 5 Fig. 4 7 Fig CCMI Blind Test Table 3 Table 3 Fig. 4 Fig. 5 Reproducibility and repeatability of calibration curve for cashmere/wool mixture. Reproducibility and repeatability of calibration curve for cashmere/yak mixture. Table 3 Peptide analysis of blind test samples* % Table 4 Table % Table 5
6 Table 4 Validation of peptide method (1). Table 5 Validation of peptide method (2). 3 4%
7 SDS-PAGE MALDI-TOF m/e= A 33 I 23 m/e= MALDI-TOF blind test DNA 2,10 Karl Spilhaus MS/MS 1. S. Subramanian, T. Karthik, N. N. Vijayaraaghavan, J. Biotechnol., 116, 153 (2005). 2. K. Kerkhoff, G. Cescutti, L. Kruse, J. Muessig, Text. Res. J., 79, 69 (2009). 3. W. Ji, L. Bai, M. X. Yang, Forensic Sci. Int., 208, 139 (2011). 4. http : / / www. meti. go. jp / information / downloadfiles / c110204d02j.pdf 5. C. Tonetti, C. Vineis, A. Aluigi, C. Tonin, Text. Res. J., 82, 766 (2012). 6. S. Ohashi, Y. Demura, M. Sano, Sen I Gakkaishi, 68, 276 (2012). 7. S. Ohashi, Y. Demura, M. Sano, Y. Yoshioka, Sen I Gakkaishi, inpress. 8. HA Ebhardt, ZZ Xu, AW Fung, RP Fahlman, Anal. Chem., 81, 1937 (2009). 9. KM Park, YJ Bae, SH Ahn, MS Kim, Anal. Chem., 84, (2012). 10. C.Vineis, C. Tonetti, S. Paolella, P.D. Pozzo, S. Sforza, Text. Res. J., December 12,
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