Bìa tạp chí



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

Pham Thi Mai Huong Vu Thi Trang Vu Thi Nhat Le Luu Thi Huyen Trang
Received: 23 Oct 2021
Revised: 23 Dec 2021
Accepted: 23 Dec 2021
Published: 30 Dec 2021

Article Details

How to Cite
Pham Thi Mai Huong, Vu Thi Trang, Vu Thi Nhat Le, Luu Thi Huyen Trang. "Determination of morpholine residues in apples by liquid chromatography tandem mass spectrophotometry (LC-MS/MS)". Vietnam Journal of Food Control. vol. 4, no. 4 (en), pp. 285-291, 2021

Main Article Content


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.


morpholine, UPLC-MS/MS, apple, apple peel, apple pulp


[1]. K. An, I. Kim, C. Lee, J. Moon, H. Suh, and J. Lee, “Quantification of Morpholine in Peel and Pulp of Apples and Oranges by Gas Chromatography - Mass Spectrometry,” Foods, vol. 9, no. 6, pp.746, 2020.
[2]. P. Cao, D. Chen, J. Liang, P. Wu, S. Wen, J. Wei, S. Liu, X. Wu, Y. Zhao, and H. Xu, “Concentration of morpholine residues in major fruits and juices and its dietary exposure in China,” Food Additives & Contaminants: Part A, vol. 36, no. 1, pp.26-34, 2019.
[3]. M. J. Walker, K. Gray, C. Hopley, D. Bell, P. Colwell, P. Maynard, and D.T. Burns, “Forensically Robust Detection of the Presence of Morpholine in Apples - Proof of Principle,” Food Analytical Methods, vol. 5, no. 4, pp. 874-880, 2011.
[4]. E. Kuchowicz and K. Rydzyn Ãåski, “Risk assessment of morpholine (tetrahydro-2H-1, 4-oxazine): A time for reevaluation of current occupational exposure standards?,” Applied Occupational and Environmental Hygiene, vol. 13, no. 2, pp.113-121, 1998.
[5]. R. Kumar and S. Kapur, “Morpholine: A Glazing Agent for Fruits and Vegeatables Coating/Waxing,” International Journal of Science Technology & Engineering, vol. 2, no. 11, pp. 694-697, 2016.
[6]. European Commission. Food Additives. “Database”. [Online]. Available: https://ec.europa.eu/food/safety/food-improvement-agents/additives/database_en. [Accessed Sept 2021].
[7]. Health Canada. “Archived-A Summary of health hazard assessment of morpholine in wax coatings of apples,”. [Online]. Available: https://www.canada.ca/en/health-canada/services/food-nutrition/food-safety/information-product/summary-health-hazard-assessment-morpholine-coatings-apples.html. [ Accessed 23/10/2021].
[8]. J. Luong, R. A. Shellie, H. Cortes, R. Gras, and T. Hayward, “Ultra-trace level analysis of morpholine, cyclohexylamine, and diethyl aminoethanol in steam condensate by gas chromatography with multi-mode inlet, and flame ionization detection,” Journal of Chromatography A, vol. 1229, pp. 223-229, 2012.
[9]. M. Cao, P. Zhang, Y. Feng, H. Zhang, H. Zhu , K. Lian , and W. Kang, "Development of a method for rapid determination of morpholine in juices and drugs by gas chromatography-mass spectrometry," Journal of Analytical Methods in Chemistry, vol 2018, pp.1-8, 2018.
[10]. L. Sabatino, M. Scordino, R. Caruso, E. Chiappara, P. Traulo, A. Belligno, and G. Gagliano, “LC/MS/MS detection of short-chain aliphatic amines in glazing agents for fruit coating,” European Food Research and Technology, vol. 235, no. 1, pp. 177-184., 2012.
[11]. M. Hengel, R. Jordan, and W. Maguire, “Development and validation of a standardized method for the determination of morpholine residues in fruit commodities by liquid chromatography-mass spectrometry,” Journal of Agricultural and Food Chemistry, vol. 62, no. 17, pp. 3697-3701, 2018.
[12]. Chen. D, Miao. H, Zou. J, Cao. P, Ma. N, Zhao. Y, Wu. Y, “Novel Dispersive Micro-Solid-Phase Extraction Combined with Ultrahigh-Performance Liquid Chromatography-High-Resolution Mass Spectrometry To Determine Morpholine Residues in Citrus and Apples,” Journal of Agricultural and Food Chemistry, vol. 63, no. 2, pp. 485-492, 2015.
[13]. AOAC Official Methods of Analysis, Guidelines for Standard Method Performance Requirements, Appendix F, 2016.