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Evidence-Based Complementary and Alternative Medicine
Volume 2012 (2012), Article ID 548486, 10 pages
http://dx.doi.org/10.1155/2012/548486
Research Article

LC-MS/MS Identification of a Bromelain Peptide Biomarker from Ananas comosus Merr

1Department of Immunology, UConn Health Center, Farmington, CT 06030-1319, USA
2The Carole and Ray Neag Comprehensive Cancer Center, UConn Health Center, Farmington, CT 06030-1628, USA
3Allergy Group, Nestlé Research Center, P.O. Box 44, 1000 Lausanne, Switzerland
4Center for Vascular Biology, UConn Health Center, Farmington, CT 06030, USA
5Department of Allied Health Sciences, University of Connecticut, Storrs, CT 06269-2101, USA

Received 25 May 2012; Accepted 2 August 2012

Academic Editor: Vassya Bankova

Copyright © 2012 Eric R. Secor Jr. et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Bromelain (Br) is a cysteine peptidase (GenBank AEH26024.1) from pineapple, with over 40 years of clinical use. The constituents mediating its anti-inflammatory activity are not thoroughly characterized and no peptide biomarker exists. Our objective is to characterize Br raw material and identify peptides in the plasma of Br treated mice. After SDS-PAGE in-gel digestion, Br (VN#3507; Middletown, CT, USA) peptides were analyzed via LC/MS/MS using 95% protein probability, 95% peptide probability, and a minimum peptide number = 5. Br spiked mouse plasma (1 ug/ul) and plasma from i.p. treated mice (12 mg/kg) were assessed using SRM. In Br raw material, we identified seven proteins: four proteases, one jacalin-like lectin, and two protease inhibitors. In Br spiked mouse plasma, six proteins (ananain, bromelain inhibitor, cysteine proteinase AN11, FB1035 precursor, FBSB precursor, and jacalin-like lectin) were identified. Using LC/MS/MS, we identified the unique peptide, DYGAVNEVK, derived from FB1035, in the plasma of i.p. Br treated mice. The spectral count of this peptide peaked at 6 hrs and was undetectable by 24 hrs. In this study, a novel Br peptide was identified in the plasma of treated mice for the first time. This Br peptide could serve as a biomarker to standardize the therapeutic dose and maximize clinical utility.