The European reference method (ERM) recognises the fraudulent addition of bovine (B) milk in water buffalo (WB) milk/dairy products based on concomitant isoelectric focusing (IEF) detection of B c2- and c3- CN fragments after corresponding plasminolysis. We here used proteomics to characterise false positive results occurring in the ERM as being due to WB b-CN(f100-209), which is also formed after plasminolysis of genuine WB milk/dairy products and comigrates in IEF with B c2-CN. These ERM limitations were overcome by a dedicated proteomic procedure based on loading of B/WB milk/cheese CN extracts on a hydroxyapatite column, in situ trypsinolysis and elution of B b-CN(f1-25)4P and WB b-CN(f1-28)4P proteotypic peptides. Based on their similar ionisation properties and resolution in MALDI-TOF-MS, these phosphopeptides were identified as suitable markers for detection of B material in WB milk/dairy products to a detection limit of 0.8% v/v. This proteomic procedure is here proposed as integrative/alternative to the ERM.
Eventual limits of the current EU official method for evaluating milk adulteration of water buffalo dairy products and potential proteomic solutions / Caira, S; Nicolai, MARIA ADALGISA; Lilla, S; Calabrese, Mg; Pinto, Gabriella; Scaloni, A; Chianese, Lina; Addeo, Francesco. - In: FOOD CHEMISTRY. - ISSN 1873-7072. - 230:(2017), pp. 482-490. [10.1016/j.foodchem.2017.03.055]
Eventual limits of the current EU official method for evaluating milk adulteration of water buffalo dairy products and potential proteomic solutions
NICOLAI, MARIA ADALGISA;PINTO, GABRIELLA;CHIANESE, LINA;ADDEO, FRANCESCO
2017
Abstract
The European reference method (ERM) recognises the fraudulent addition of bovine (B) milk in water buffalo (WB) milk/dairy products based on concomitant isoelectric focusing (IEF) detection of B c2- and c3- CN fragments after corresponding plasminolysis. We here used proteomics to characterise false positive results occurring in the ERM as being due to WB b-CN(f100-209), which is also formed after plasminolysis of genuine WB milk/dairy products and comigrates in IEF with B c2-CN. These ERM limitations were overcome by a dedicated proteomic procedure based on loading of B/WB milk/cheese CN extracts on a hydroxyapatite column, in situ trypsinolysis and elution of B b-CN(f1-25)4P and WB b-CN(f1-28)4P proteotypic peptides. Based on their similar ionisation properties and resolution in MALDI-TOF-MS, these phosphopeptides were identified as suitable markers for detection of B material in WB milk/dairy products to a detection limit of 0.8% v/v. This proteomic procedure is here proposed as integrative/alternative to the ERM.| File | Dimensione | Formato | |
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