Simultaneous Immunoaffinity Column Cleanup and HPLC Analysis of Aflatoxins and Ochratoxin A in Spanish Bee Pollen

Abstract

Bee pollen is a major substrate for mycotoxins growth when no prompt and adequate drying is performed by the beekeeper after collection by bees. Regulatory limits for aflatoxins and ochratoxin A are currently in force in the European Union for a rising list of foodstuffs, but not for this. An immunoaffinity column cleanup process has been applied prior to the analysis of aflatoxins B(1), B(2), G(1), and G(2) and ochratoxin A (OTA). Optimization of the HPLC conditions has involved both a gradient elution and a wavelength program for the separation and fluorimetric quantitation of all five mycotoxins at their maximum excitation and emission values of wavelength in a single run. The higher limit of detection (mug/kg) was 0.49 for OTA and 0.20 for aflatoxin B(1). Repeatability (RSDr) at the lower limit tested ranged from 9.85% for OTA to 6.23% for aflatoxin G(2), and recoveries also at the lower spiked level were 73% for OTA and 81% for aflatoxin B(1). None of the 20 samples assayed showed quantifiable values for the five mycotoxins.

HPLC Method Validation for Simultaneous Determination of Three Mycotoxins from Corn Seeds

“…The lower limits of quantification LOQ for the analyzed species were determined with the following values: 2.88 µg kg -1 aflatoxin B1, 2.88 µg kg -1 ochratoxin A respectively 14.4 µg kg -1 zearalenone and the records have respected the imposed conditions required by the GLP regulations (meaning that relative standard deviation should not exceed ±20 % also the bias in the same domain). 23,24 The quantification limits that were published and reached with complex instruments are in a large range between 0.25 /16 µg kg -1 aflatoxin B1, [15][16][17][18] 0.25 /8.0 µg kg -1 ochratoxin A, 15-17 10/40 µg kg -1 for zearalenone, 15,25 mostly depending not only on the performance of the instruments, but on the extraction procedure applied for each type. Comparing our limits of quantification mentioned above for the three mycotoxins, one can see that they are in the ranges of these values presented lately by other groups, even we used a simple HPLC system.…”

Study of aflatoxin B1 production by Aspergillus parasiticus in bee pollen of Greek origin

“…In the present study, the lower LOD=0.08 ng AFB 1 /g was utilized throughout this work. Garcia-Villanova et al (2004) determined AFB 1 and ochratoxin A in Spanish bee pollen. The recovery factor and LOD for AFB 1 were found to be 81% and 0.20 ng/g of pollen, respectively.…”

Ochratoxin A quantification: Newly developed HPLC conditions

“…In tropical regions, Aspergillus species are the main source of ochratoxin contamination in foods and feeds (Garcia-Villanova, Cordn, Gonzlez Params, Aparicio, & Garcia Rosales, 2004). OTA contamination can also be transferred from feed into animal products, especially in organ meat (kidney, liver, and blood).…”

“…OTA has been associated with nephropathy in humans from the Balkan area (Balkan Endemic Nephropathy; BEN), and it has also been associated with the occurrence of kidney tumors (Ceovic, Hrabar, & Saric, 1992;Garcia-Villanova, et al, 2004). In North African countries, similar disease symptoms with a high incidence of chronic interstitial nephropathies of unknown etiology have been observed, and OTA was suspected to have a role in these nephropathies, especially in Morocco, Tunisia and Algeria.…”