Selective and rapid immunomagnetic bead-based sample treatment for the liquid chromatography-electrospray ion-trap mass spectrometry detection of Ara h3/4 peanut protein in foods.
Complex matrices commonly affect the sensitivity and selectivity of liquid chromatography-mass spectrometry (LC-MS) analysis. Thus, selective sample enrichment strategies are useful particularly to analyze organic biomarkers present in low abundance in samples. A selective immunomagnetic extraction procedure to isolate trace peanut allergen protein Ara h3/4 from breakfast cereals combined with microwave-assisted tryptic digestion and liquid chromatography-electrospray ion-trap tandem mass spectrometry (LC-ESI-IT-MS/MS) measurement was developed. Using protein A-coated magnetic bead (MB) support, anti-Ara h3/4 monoclonal antibodies (Abs) were used as selective capture molecules. The results obtained by LC-ESI-IT-MS/MS in terms of limit of detection (3 mg peanuts/kg matrix) and a significantly reduced matrix effect demonstrated that the Ab-coated magnetic bead was very effective to selectively trap Ara h3/4 protein in breakfast cereals. The magnetic bead-based sample treatment followed by LC-IT-MS/MS method here developed can be proposed as very rapid and powerful confirmatory analytical method to verify the reliability of enzyme-linked immunosorbent assay (ELISA) screening methods, since the magnetic bead-LC-IT-MS/MS method combines good sensitivity to the identification capabilities of mass spectrometry.
Careri
Maria
M
Dipartimento di Chimica Generale ed Inorganica, Chimica Analitica, Chimica Fisica, Università degli Studi di Parma, Viale G.P. Usberti 17/A, I-43100 Parma, Italy.
Elviri
Lisa
L
Lagos
Jesse Boquett
JB
Mangia
Alessandro
A
Speroni
Francesca
F
Terenghi
Mattia
M
D000485:Allergens / Q000032:analysis*; D052179:Antigens, Plant; D002853:Chromatography, Liquid / Q000379:methods*; D005504:Food Analysis; D006023:Glycoproteins / Q000032:analysis*; D018189:Immunomagnetic Separation / Q000379:methods*; D010940:Plant Proteins / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055314:Seed Storage Proteins
D000485:Allergens; D052179:Antigens, Plant; D006023:Glycoproteins; D010940:Plant Proteins; C118207:allergen Ara h3; D055314:Seed Storage Proteins