Determination of morpholine residues in apples by liquid chromatography tandem mass spectrophotometry (LC-MS/MS)

Morpholine is an emulsifier that is normally used as a protective wax coating on the surfaces of fruit and vegetable. In order to keep fruits fresh, extend their shelf-life, against insect and fungal contamination for a long time, morpholine has been widely applied despite of the reported carcinogenic potential in previous studies and export prohibition in many countries. A highly sensitive method has been developed for the determination of this food additive in fruits using ultra-performance liquid chromatography tandem mass spectrophotometry. By using 1 % acetic acid in methanol for extraction and analyzing with binary ammonium formate 20 mM and acetonitrile, the method has been applied successfully to determine morpholine residues in apple samples collected at local markets in Hanoi, Vietnam. The good coefficient correlation was achieved in the linear range of 5 - 300 µg/L (R2 = 0.9998). The low limit of detection (LOD) and limit of quantification (LOQ) were 2 µg/kg and 5 µg/kg, respectively. The recoveries of morpholine were 83 – 108 % over three spike levels 20 - 60 μg/L for apple, peel and pulp matrices. The repeatability (RSDr %) was 1.1 % for apple matrix, 2.36 % for peel and 3.67 % for pulp. The reproducibility values (RSDR %) were 1.49, 3.50 and 4.08 % for apple, peel and pulp, respectively. This method could be further performed in different type of fruit and vegetable comiditities to determine the morpholine content.