Saffron Agronomy and Technology

Saffron Agronomy and Technology

Identification of Apo- Carotenoids' Crocin and Crocetin Isomers in Saffron Crude Extracts by HPLC Coupled to Atmospheric Pressure Chemical Ionization and High Resolution Orbitrap Mass Spectrometry

Document Type : Research Paper

Authors
Noraddin Hosseinpour azad 1
Ghorban Ali Nematzadeh 2
Giovanni Giuliano 3
Gholam Ali Ranjbar 4
Ahad Yamchi 5
1 Assist. Prof. of Planr breeding-Genetic Engineering, Meshkinshahr Faculty of Agriculture, University of Mohaghegh Ardabili, Meshkinshahr, Iran.
2 Academic member at Department of Plant Breeding, Sari University of Agriculture, Sari, Iran
3 Prof., Plant Biology, Department of Biotechnology, Italian National Agency for New Technologies, Rome, Italy.
4 Academic member at Department of Plant Breeding, Sari University of Agriculture, Sari, Iran.
5 Assist. Prof. of Genetic Engineering and Molecular Genetics,Gorgan University of Agricultural Science and Natural Resources Golestan, Gorgan, Iran,
10.22048/jsat.2016.38670
Abstract
The main metabolites in saffron are the Apo- carotenoids’ Crocin and Crocetin. Color intensity and quality of saffron mostly depend on the presence of Crocins that are glycosylated steric form of Crocetin by glycosyltransferase enzyme. The aim of this study is the characterization of these metabolites in methanolic and chloroformic extracts of saffron stigmas during anthesis stage by LC-APCI-MS. Identification of cis and trans isomers of Crocin and Crocetin was done by three parameters such as mass spectra registered in the negative ion mode, retention time and absorption ratio related to each metabolites. The variability of these parameters made it possible to detect the Crocins isomer with regard to the attached position and the number of UDP- glucose and Gentiobiosyl molecules to Crocetin structure. Crocins was the mainly detected components as there are polar components that are classified in the carotenoeids groups and the strified form of Crocetin Glucose (β-D-Glucopyranosyl) and Gentiobiose (β-D-Glucopyranosyl-D-Glucose). Also doubly charged ions were found for trans-isomers of Crocin-4, due to the high symmetry of their molecules. Based on the data gathered, the applied chromatograph Machin in this project is accurate and it is most sensitive tools to investigate about plants’ natural components like saffron, also the used APCI-MS in negative ions mode is the most efficient method to distinguish different steric forms of Crocin based on the ion’s fragments related to united reduction of glycosyl and gentiobiosyl as well as molecular fractions.
Keywords
Saffron
Metabolite
Crocetin
Crocin
Mass spectrometry

Subjects

Caballero-Ortega, H., Pereda-Miranda, R., and Abdullaev, F.I. 2007. HPLC quantification of major active components from 11 different saffron (Crocus sativus L.) sources. Food Chemistry 100 (3): 1126–1131.
Carmona, M., Zalacain, A., Saa N., A.M., Novella, J.L., and Alonso, G.L. 2006. Crocetin esters, Picrocrocin and its related compounds present in Crocus sativus stigmas and gardenia jasminoides fruits. Tentative Identification of Seven New Compounds by LC-ESI-MS. Journal of Agricultural Food Chemistry 54: 973-979.
Casas-Catal´an, M.T., and Dom´enech-Carb´, O. 2005. Identificationofnatural dyes used in works of art by pyrolysis-gas chromatography/mass spectrometry combined with in situ trimethylsilylation. Analytical and Bioanalytical Chemistry 382: 259-268.
Dauria, M., Mauriello, G., Racioppi, R., and Rana, G.L. 2006. Use of SPME-GC–MS in the study of time evolution of the constituents of saffron aroma, modifications of the composition during storage. Journal of Chromatographic Science 44: 18–21.
Esmaeilian, Y., Galavi, M., Ramroudi, M., and Boojar, M.M.A. 2012. Diurnal variability of stigma compounds of saffron (Crocus sativus L.) at different harvest times. Annals of Biological Research journal 3 (3):1562-1568.
Fantini, E., Falcone, G., Frusciante, S., Giliberto, L., and Giuliano G. 2013. Dissection of tomato Lycopene biosynthesis through virus-Induced gene silencing. Plant Physiology 163: 986-998.
Fernandez, J.A., and Pandalai, S.G. 2004. Biology,
    biotechnology and biomedicine of saffron. Recent Research Developement Plant Science
   2: 127–159.
Goli, S.A.H., Mokhtari, F., and Rahimmalek, M. 2012. Phenolic Compounds and Antioxidant Activity from Saffron (Crocus sativus L.) Petal. Journal of Agricultural Science 4 (10).
Gomez-Gomez, L., Moraga, A.R., and Ahrazen, O. 2010. Understanding carotenoid metabolism in saffron stigmas, Unraveling aroma and color formation. Functional Plant Science 4: 56-63.
Kanakis, C.D., Daferera, D.J., Tarantilis, P.A., and Polissiou, M.G. 2004. Qualitative determination of volatile compounds and quantitative evaluation of safranal and 4-hydroxy-2, 6, 6-trimethyl-1-cyclohexene-1-carboxaldehyde (HTCC) in Greek saffron, Journal of Agricultural Food Chemistry 52: 4515–4521.
Tarantilis, P.A., Tsoupras, G., and Polissiou, M. 1995. Determination of saffron (Crocus sativus L.) components in crude plant extract using highperformance liquid chromatography-UV-visible photodiode-array detection-mass spectrometry. Journal of Chromatography 699: 107-117.
Lozano, P., Castellar, M.R., Simancas, M.J., and Iborra, J.L. 1999. A quantitative high-performance liquid chromatographic method to analyse commercial saffron (Crocus sativus L.) products. Journal of Chromatography 830: 477-483.
Jin, R.L., Qiao, Z.P., Zhou, S.D., Ye, Y.L., and Zhou, J.X. 1986. Investigation of saffron preparations by thin layer chromatography. Journal of Nanjing College of Pharmacy 17: 247.
Rubio, A., Rambla, J.L., Santoella, M., Gomez, M.D., Orzaez, D., Granell, A., and Gomez-Gomez, L. 2008. Cytosolic and plastoglobule targeted caroteniod dioxygenase from Crocus sativus are both involved in β-ionone release. Journal of Biology Chemistry 283: 24816-24825.
Saffron Agronomy and Technology
Volume 4, Issue 4 - Serial Number 14
Winter 2017
Pages 291-300