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Determination of meaty peptide in enzymatic hydrolyzate of beef protein by HPLC-MS
Yanping Wang,Songrong Zeng,Xiaojia Bai,Weili Lin,Ming Yang,Haipeng Xing 한국화학공학회 2008 Korean Journal of Chemical Engineering Vol.25 No.5
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
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