Determination of meaty peptide in enzymatic hydrolyzate of beef protein by HPLC-MS

Songrong Zeng; Yanping Wang; Xiaojia Bai; Weili Lin; Ming Yang; Haipeng Xing

Korean Journal of Chemical Engineering, Vol.25, No.5, 1120-1124, September, 2008

DOI10.1007/s11814-008-0183-2 Export Citation

Determination of meaty peptide in enzymatic hydrolyzate of beef protein by HPLC-MS

Songrong Zeng^{1, 2}, Yanping Wang^{1, †}, Xiaojia Bai¹, Weili Lin¹, Ming Yang¹, and Haipeng Xing³

¹College of Food Engineering & Biotechnology, Tianjin Key Laboratory of Food Nutrition & Safety, Tianjin University of Science & Technology, Tianjin 300457, China
²Yingdong College of Bioengineering, Shaoguan University, Shaoguan 512005, China
³Tianjin Chunfa Food Ingredient Limited Company, Tianjin, 300300, China

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The purpose of this study is to detect beefy meaty peptide (BMP) in beef hydrolyzate. The synthesized BMP is used as a standard sample in the study. High performance liquid chromatography (HPLC)/ion trap electrospray ionization mass spectrometry (ESI-MS) with UV detection was applied in qualitative analysis of the peptides. Six beef protein enzymatic hydrolyzate samples were separated on a Surveyor HPLC system through a SUPELCO Discovery® C18 analytical column (5 μm, 15 cm×2.1 mm i.d.). The column was eluted at a flow rate of 0.2 mL/min in a linear gradient elution mode of acetronitrile-water solution with 0.1% trifluoroacetic acid. The concentration of acetronitrile was increased from 5% to 50% in 40 minutes. A Finnigan LCQ Advantage MAX instrument was used as detector to analyze with ESI-MS and ESI-MS/MS in positive mode. Among the six samples of beef protein enzymatic hydrolysate, the BMP is detected and confirmed in sample No.4 with a higher intensity of characteristic peak and is further investigated by ESI-MS/MS. As a result, BMP exists in sample No.4. The study proves that HPLC-ESI-MS/MS is a simple, rapid, sensitive method to analyze target peptides from complex polypeptides.

Keywords:Beef;Enzymatic Hydrolysis;Beefy Meaty Peptide (BMP);HPLC-MS

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[1] Yamasaki Y, Maekawa K, Agric. Biol. Chem., 42, 1761 (1978)

[2] Wang K, Maga JA, Bechtel PJ, J. Food Sci., 61, 837 (1996)

[3] Wang K, Maga JA, Bechtel PJ, Lebensm.-Wiss. u.-Technol, 28, 539 (1995)

[4] Gomez-Ruiz JA, Ramos M, Recio I, J. Chromatogr. A, 1054, 269 (2004)

[5] Lee YW, Jin YZ, Row KH, Korean J. Chem. Eng., 23(2), 279 (2006)

[6] Cui LJ, Nithipatikom K, Campbell WB, Anal. Biochem., 369, 27 (2007)

[7] Gorman JJ, Wallis TP, Pitt JJ, Mass Spectrum. Rev., 21, 183 (2002)

[8] Yamashita K, Okuyama M, Watanabe Y, Honmab S, Kobayashia S, Numazawaa M, Steroids, 72, 829 (2007)

[9] Tsimogiannis D, Samiotaki M, Panayotou G, Oreopoulou V, Molecules, 12, 593 (2007)

[10] Favretto D, Frison G, Maietti S, Ferrara SD, Int. J. Legal Med., 121, 259 (2007)