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| HPLC determination of ochratoxin A in roasted coffee beans |  |
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DELIA BULEA1, ADRIAN ŞPAC2, VASILE DORNEANU3
(1) Disciplina Chimia mediului şi alimentului, Facultatea de Farmacie, Universitatea de Medicină şi Farmacie „Gr. T. Popa” Iaşi
(2) Disciplina Chimie - fizică, Facultatea de Farmacie, Universitatea de Medicină şi Farmacie „Gr. T. Popa” Iaşi
(3) Disciplina Chimie analitică, Facultatea de Farmacie, Universitatea de Medicină şi Farmacie „Gr. T. Popa” Iaşi
(2) Disciplina Chimie - fizică, Facultatea de Farmacie, Universitatea de Medicină şi Farmacie „Gr. T. Popa” Iaşi
(3) Disciplina Chimie analitică, Facultatea de Farmacie, Universitatea de Medicină şi Farmacie „Gr. T. Popa” Iaşi
Aim: In the present paper we proposed to validate a high performance liquid chromatographic (HPLC) analysis method for ochratoxin A determination and to apply the method for roasted coffee beans.
Material and method: The samples of roasted coffee beans were purchased in the Iasi markets. The sample was homogenized with CH3OH:NaHCO3 (80:20) mixture. Sample extract was cleanup by SPE extraction. Ochartoxin A was eluted with chloroform. Organic phase was dried and the residue was redissolved in 0.6 ml mobile phase; 20 μl was injected in the HPLC sistem. The HPLC analysis was performed to an HP 1090 chromatograph equiped with both diode array detector and fluorescence detector. The stationary phase is a Stable Bond SB - C18 column (150 x 4.6 mm; 5 μm). The mobile phase (0.7 ml/min) was a mixture of CH3CN : H2O : CH3COOH (99 : 99 : 2). The volume of injected sample is 20 μl. The detection was performed in fluorescence (excitation at 228 nm and emmision at 423 nm).
Material and method: The samples of roasted coffee beans were purchased in the Iasi markets. The sample was homogenized with CH3OH:NaHCO3 (80:20) mixture. Sample extract was cleanup by SPE extraction. Ochartoxin A was eluted with chloroform. Organic phase was dried and the residue was redissolved in 0.6 ml mobile phase; 20 μl was injected in the HPLC sistem. The HPLC analysis was performed to an HP 1090 chromatograph equiped with both diode array detector and fluorescence detector. The stationary phase is a Stable Bond SB - C18 column (150 x 4.6 mm; 5 μm). The mobile phase (0.7 ml/min) was a mixture of CH3CN : H2O : CH3COOH (99 : 99 : 2). The volume of injected sample is 20 μl. The detection was performed in fluorescence (excitation at 228 nm and emmision at 423 nm).
Results: The HPLC method was validated. Ochratoxin A has been found in coffee beans as follows: in 12.5% from the samples, ochratoxin A concentration is under the limit value (5 μg/kg roasted coffee beans), in 2.5% from the samples ochratoxin A exceded the limit value, and in 85% from the sample ochratoxin A is below the limit of detection.
Discussion: This research contributes to the evaluation of the presence of ochratoxin A in Romanian food and draw attention upon the risk of consumption of food containing mycotoxins.
Discussion: This research contributes to the evaluation of the presence of ochratoxin A in Romanian food and draw attention upon the risk of consumption of food containing mycotoxins.
Keywords:
HPLC, ochratoxina A, cafea boabe prăjită