10.57647/tpr.2025.0904.19

Chemical composition of the essential oils and volatilefractions from the aerial parts of Crocus sativus L.(Iridaceae) using microwave assisted hydrodistillation (MAHD) and headspace solid phase microextraction(HS-SPME) combined with gas chromatography-massspectrometric (GC-MS) analysis

  1. Herbal Drugs Raw Materials Research Center (HDRMRC), Sha.C., Islamic Azad University, Shahrood, Iran
  2. Department of Chemistry and Biochemistry, Sha.C., Islamic Azad University, Shahrood, Iran

Received: 2025-08-02

Revised: 2025-11-27

Accepted: 2025-12-05

Published in Issue 2025-12-31

How to Cite

Mohammadhosseini, M. (2025). Chemical composition of the essential oils and volatilefractions from the aerial parts of Crocus sativus L.(Iridaceae) using microwave assisted hydrodistillation (MAHD) and headspace solid phase microextraction(HS-SPME) combined with gas chromatography-massspectrometric (GC-MS) analysis. Trends in Phytochemical Research, 9(4). https://doi.org/10.57647/tpr.2025.0904.19

PDF views: 2

Abstract

In this report, chemical composition of the essential oils and volatile fractions from the aerial parts of Crocus sativus L. (Iridaceae) was investigated using microwave-assisted hydrodistillation (MAHD) and headspace solid-phase microextraction (HS-SPME), followed by gas chromatography-mass spectrometry (GC-MS) analysis on DB-5 and HP-5MS columns. Twenty constituents were identified by matching their mass spectra, retention indices, and co-injection with reference standards. In the MAHD-derived essential oil, seventeen compounds accounted for 97.3% of the total composition, with safranal (37.2%) as the major component, followed by 4-ketoisophorone (11.2%), hexadecane (9.1%), hexadecanoic acid (8.1%), tetradecanoic acid (7.5%), nonacosane (6.2%), and n-eicosane (5.1%). The HS-SPME volatile fraction exhibited a higher safranal content (63.5%), accompanied by hexadecanoic acid (10.2%), ethyl hexadecanoate (6.7%), tetradecanoic acid (5.7%), and isophorone (3.0%). These findings reveal pronounced method-dependent variations in the volatile profiles, underscoring the dominance of safranal and highlighting the potential of saffron aerial parts as an alternative source of valuable aroma compounds.

Keywords

  • Crocus sativus L.,
  • Iridaceae; Microwave Assisted Hydrodistillation (MAHD),
  • Headspace Solid Phase Microextraction (HS-SPME),
  • Volatile fractions,
  • Safranal (2,6,6-Trimethyl-1,3-cyclohexadien-l-carboxaldehyde)

References

  1. Amanpour, A., Sonmezdag, A.S., Kelebek, H., Selli, S. (2015) GC-MS-olfactometric characterization of the most aroma-active components in a representative aromatic extract from Iranian saffron (Crocus sativus L.). Food Chem. 182:251–256. DOI: https://doi.org/10.1016/j.foodchem.2015.03.005.
  2. Anabat, M.M., Riahi, H., Sheidai, M., Koohdar, F. (2020) Population genetic study and barcoding in Iran saffron (Crocus sativus L.). Ind. Crops Prod. 143. DOI: https://doi.org/10.1016/j.indcrop.2019.111915.
  3. Azarabadi, N., O¨ zdemir, F. (2018) Determination of crocin content and volatile components in different qualities of Iranian saffron. Gıda 43(3):476–489. DOI: https://doi.org/10.15237/gida.GD18018.
  4. Bahmani, M., Rafieian, M., Baradaran, A., Rafieian, S., Rafieian-Kopaei, M. (2014) Nephrotoxicity and hepatotoxicity evaluation of Cro-cus sativus stigmas in neonates of nursing mice. J. Nephropathol. 3(2):81–85. DOI: https://doi.org/10.12860/jnp.2014.16.
  5. Bajbouj, K., Schulze-Luehrmann, J., Diermeier, S., Amin, A., Schneider-Stock, R. (2012) The anticancer effect of saffron in two p53 iso-genic colorectal cancer cell lines. BMC Complement. Altern. Med. 12:1100. DOI: https://doi.org/10.1186/1472-6882-12-69.
  6. Basheeruddin Asdaq, S.M.B., Inamdar, M.N. (2010) Potential of Crocus sativus (saffron) and its constituent, crocin, as hypolipidemic and antioxidant in rats. Appl. Biochem. Biotechnol. 162(2):358–372. DOI: https://doi.org/10.1007/s12010-009-8740-7.
  7. Boozari, M., Hosseinzadeh, H. (2022) Crocin molecular signaling pathways at a glance: A comprehensive review. Phytother. Res. 36(10):3859–3884. DOI: https://doi.org/10.1002/ptr.7583.
  8. Boskabady, M.H., Farkhondeh, T. (2016) Antiinflammatory, antioxidant, and immunomodulatory effects of Crocus sativus L. and its main constituents. Phytother. Res. 30(7):1072–1094. DOI: https://doi.org/10.1002/ptr.5622.
  9. Cerda´-Bernad, D., Clemente-Villalba, J., Valero-Cases, E., Pastor, J.J., Frutos Fernandez, M.J. (2022) Novel insight into the volatile profile and antioxidant properties of Crocus sativus L. flowers. Antioxidants 11(9). DOI: https://doi.org/10.3390/antiox11091650.
  10. Chen, D., Xing, B., Yi, H., Li, Y., Zheng, B., Wang, Y., Shao, Q. (2020) Effects of different drying methods on appearance, microstructure, bioactive compounds and aroma compounds of saffron (Crocus sativus L.). LWT 120. DOI: https://doi.org/10.1016/j.lwt.2019.108913.
  11. Condurso, C., Cincotta, F., Tripodi, G., Verzera, A. (2017) Bioactive volatiles in Sicilian (South Italy) saffron: Safranal and its related compounds. J. Essent. Oil Res. 29(3).221–227. DOI: https://doi.org/10.1080/10412905.2016.1244115.
  12. Criado-Navarro, I., Ledesma-Escobar, C.A., Pe´rez-Juan, P., Priego-Capote, F. (2024) Distribution of main bioactive compounds from saffron species as a function of infusion temperature and time in an oil/water system. Molecules 29(13). DOI: https://doi.org/10.3390/molecules29133080.
  13. D’Auria, M., Mauriello, G., Rana, G.L. (2004) Volatile organic compounds from saffron. Flavour Fragrance J. 19(1):17–23. DOI: https://doi.org/10.1002/ffj.1266.
  14. Djenhi, F., Bensouici, C., Kechebar, M.S.A., Fernandez, M.J.F., Atoki, A.V., Karoune, S., Boumechhour, A., Mustapha, M.A., Saadoun, S., Hamdi, M., Khattabi, L., Chouh, A., Messaoudi, M. (2025) Variation of the profile’s volatile components and in silico modeling of the non-polar extracts of the petals and stigmas of Algerian saffron cultivar. Nat. Pro. Comm. 20(6). DOI: https://doi.org/10.1177/1934578X251350086.
  15. Drioiche, A., Ailli, A., Handaq, N., Remok, F., Elouardi, M., Elouadni, H., Al kamaly, O., Saleh, A., Bouhrim, M., Elazzouzi, H., El Makhoukhi, F., Za¨ır, T. (2023) Identification of compounds of Crocus sativus by GC-MS and HPLC/UV-ESI-MS and evaluation of their antioxidant, antimicrobial, anticoagulant, and antidiabetic properties. Pharma-ceuticals 16(4):545. DOI: https://doi.org/10.3390/ph16040545.
  16. F.Abdel-Rahman, R.F., El Awdan, S.A., Hegazy, R.R., Mansour, D.F., Ogaly, H.A., Elbaset, M. (2020) Neuroprotective effect of Cro-cus sativus against cerebral ischemia in rats. Metab. Brain Dis. 35(3):427–439. DOI: https://doi.org/10.1007/s11011-019-00505-1.
  17. Frusciante, L., Geminiani, M., Shabab, B., Olmastroni, T., Scavello, G., Rossi, M., Mastroeni, P., Nyong’a, C.N., Salvini, L., Lamponi, S., Parisi, M.L., Sinicropi, A., Costa, L., Spiga, O., Trezza, A., Santucci, A. (2024) Exploring the antioxidant and anti-inflammatory potential of Saffron (Crocus sativus) tepals extract within the circular bioe-conomy. Antioxidants 13(9). DOI: https://doi.org/10.3390/antiox13091082.
  18. Ghanbari, J., Khajoei-Nejad, G., Erasmus, S.W., Van Ruth, S.M. (2019) Identification and characterisation of volatile fingerprints of saffron stigmas and petals using PTR-TOF-MS: Influence of nutritional treatments and corm provenance. Ind. Crops Prod. 141. DOI: https://doi.org/10.1016/j.indcrop.2019.111803.
  19. Goldblatt, P., Manning, J., Rudall, P. (1998) Iridaceae, Flowering Plants· Monocotyledons: Lilianae (Except Orchidaceae). Springer, 295–333.
  20. Goupy, P., Abert Vian, M.A., Chemat, F., Caris-Veyrat, C. (2013) Iden-tification and quantification of flavonols, anthocyanins and lutein diesters in tepals of Crocus sativus by ultra performance liquid chro-matography coupled to diode array and ion trap mass spectrometry detections. Ind. Crops Prod. 44:496–510. DOI: https://doi.org/10.1016/j.indcrop.2012.10.004.
  21. Guclu, G., Kelebek, H., Selli, S. (2020) affron (Crocus sativus L.): Its Aroma and Key Odorants. Elsevier, 682.
  22. Hamidian, M., Movahhedi Dehnavi, M., Sayyed, R.Z., Almalki, W.H., Ga-fur, A., Fazeli-Nasab, B. (2023) Co-application of Mycorrhiza and methyl jasmonate regulates morpho-physiological and antioxidant responses of Crocus sativus (Saffron) under salinity stress condi-tions. Sci. Rep. 13(1):7378. DOI: https://doi.org/10.1038/s41598-023-35118-3.
  23. Hoshyar, R., Bathaie, S.Z., Sadeghizadeh, M. (2013) Crocin triggers the apoptosis through increasing the Bax/Bcl-2 ratio and caspase activa-tion in human gastric adenocarcinoma, AGS, cells. DNA Cell Biol. 32(2):50–57. DOI: https://doi.org/10.1089/dna.2012.1866.
  24. Jalali-Heravi, M., Parastar, H., Ebrahimi-Najafabadi, H. (2009) Characteri-zation of volatile components of Iranian saffron using factorial-based response surface modeling of ultrasonic extraction combined with gas chromatography-mass spectrometry analysis. J. Chromatogr. A 1216(33):6088–6097. DOI: https://doi.org/10.1016/j.chroma.2009.06.067.
  25. Karabagias, I.K., Koutsoumpou, M., Liakou, V., Kontakos, S., Kontominas, M.G. (2017) Characterization and geographical discrimination of saffron from Greece, Spain, Iran, and Morocco based on volatile and bioactivity markers, using chemometrics. Eur. Food Res. Technol. 243(9):1577–1591. DOI: https://doi.org/10.1007/s00217-017-2866-6.
  26. Khazdair, M.R., Boskabady, M.H., Hosseini, M., Rezaee, R., Tsatsakis, A.M. (2015) The effects of Crocus sativus (saffron) and its con-stituents on nervous system: A review. Avicenna J. Phytomedicine 5(5):376.
  27. Kos¸ar, M., Demirci, B., Go¨ger, F., Kara, I., Bas¸er, K.H.C. (2017) Volatile composition, antioxidant activity, and antioxidant components in saffron cultivated in Turkey. Int. J. Food Prop. 20:S746–S754. DOI: https://doi.org/10.1080/10942912.2017.1311341.
  28. Kothari, D., Thakur, R., Kumar, R. (2021) Saffron (Crocus sativus L.): Gold of the spices–A comprehensive review. Hortic. Environ. Biotechnol. 62(5):661–677. DOI: https://doi.org/10.1007/s13580-021-00349-8.
  29. Masuda, A., Mori, K., Miyazawa, M. (2012) Comparative analysis of volatile compounds from corms of Crocus sativus and C. vernus. Chem. Nat. Compd. 48(2):319–321. DOI: https://doi.org/10.1007/s10600-012-0236-y.
  30. Nader, M., Chahine, N., Salem, C., Chahine, R. (2016) Saffron (Cro-cus sativus) pretreatment confers cardioprotection against ischemia-reperfusion injuries in isolated rabbit heart. J. Physiol. Biochem. 72(4):711–719. DOI: https://doi.org/10.1007/s13105-016-0510-8.
  31. Naim, N., Guirrou, I., Fauconnier, M.L., Hafida, H., Tahiri, A., Madani, I., Lahlali, R., Ennahli, S. (2023) Chemical, biochemical and volatile profiles of saffron (Crocus sativus L.) from different growing areas of Morocco. JSFA Rep. 3(5):233–247. DOI: https://doi.org/10.1002/jsf2.114.
  32. Rasmi, Y., Salazar, E., Gupta, E., Daei-Hasani, B., Caldero´n-Jua´rez, M. (2022) Saffron: A Functional Food with Potential Molecular Effects. Wiley, 455–484.
  33. Rodriguez ,A. (1999) Molecular and Morphological Systematics of the ”Tiger-Flower” Group (Tribe Tigridieae: Iridaceae), Biogeography and Evidence for The Adaptive Radiation of The Subtribe Tigridi-inae. The University of Wisconsin-Madison.
  34. Roshanravan, N., Ghaffari, S. (2022) The therapeutic potential of Crocus sativus Linn.: A comprehensive narrative review of clinical trials. Phytother. Res. 36(1):98–111. DOI: https://doi.org/10.1002/ptr.7286.
  35. Saadat, S., Ghasemi, S.Z., Memarzia, A., Behrouz, S., Aslani, M.R., Bosk-abady, M.H. (2024) An overview of pharmacological effects of Cro-cus sativous and its constituents. Iran. J. Basic Med. Sci. 27(4):391–417. DOI: https://doi.org/10.22038/IJBMS.2023.73410.15950.
  36. Saeidnia ,S. (2012) Future position of Crocus sativus as a valuable medici-nal herb in phytotherapy. Pharmacogn J. 4(27):71. DOI: https://doi.org/10.5530/pj.2012.27.12.
  37. Sereshti, H., Heidari, R., Samadi, S. (2014) Determination of volatile components of saffron by optimised ultrasound-assisted extraction in tandem with dispersive liquid-liquid microextraction followed by gas chromatography-mass spectrometry. Food Chem. 143:499–505. DOI: https://doi.org/10.1016/j.foodchem.2013.08.024.
  38. Sevindik ,B. (2020) Stability of volatile compounds of Turkish saffron (Crocus sativus) after one-year storage. J. Raw Mater. Process. Foods 1(2):72–79.
  39. Shao, Q., Huang, Y., Zhou, A., Guo, H., Zhang, A., Wang, Y. (2014) Application of response surface methodology to optimise supercriti-cal carbon dioxide extraction of volatile compounds from Crocus sativus. J. Sci. Food Agric. 94(7):1430–1436. DOI: https://doi.org/10.1002/jsfa.6435.
  40. Siddiqui, M.J., Saleh, M.S., Basharuddin, S.N.B., Zamri, S.H.B., bin Mohd Najib, M.H., bin Che Ibrahim, M.Z., Mazha, H.N.B., Hassan, N.M., Khatib, A. (2018) Saffron (Crocus sativus L.): As an antidepressant.
  41. J. Pharm. Bioallied Sci. 10(4):173–180. DOI: https://doi.org/10.4103/jpbs.JPBS“˙83“˙18.
  42. Singab, A.N.B., Ayoub, I.M., El-Shazly, M., Korinek, M., Wu, T.Y., Cheng, Y.B., Chang, F.R., Wu, Y.C. (2016) Shedding the light on Iridaceae: Ethnobotany, phytochemistry and biological activity. Ind. Crops Prod. 92:308–335. DOI: https://doi.org/10.1016/j.indcrop.2016.07.040.
  43. Srivastava, R., Ahmed, H., Dixit, R., Saraf, S. (2010) Crocus sativus L.: A comprehensive review. Pharmacogn. Rev. 4(8):200–208. DOI: https://doi.org/10.4103/0973-7847.70919.
  44. Tarantilis, P.A., Polissiou, M.G. (1997) Isolation and identification of the aroma components from saffron (Crocus sativus). J. Agric. Food Chem. 45(2):459–462. DOI: https://doi.org/10.1021/jf960105e.
  45. Tirillini, B., Pagiotti, R., Menghini, L., Miniati, E. (2006) The volatile or-ganic compounds from tepals and anthers of saffron flowers (Crocus sativus L.). J. Essent. Oil Res. 18(3):298–300. DOI: https://doi.org/10.1080/10412905.2006.9699095.
  46. Wali, A.F., Alchamat, H.A.A., Hariri, H.K., Hariri, B.K., Menezes, G.A., Zehra, U., Rehman, M.U., Ahmad, P. (2020) Antioxidant, antimicro-bial, antidiabetic and cytotoxic activity of Crocus sativus L. petals. Appl. Sci. 10(4). DOI: https://doi.org/10.3390/app10041519.
  47. Wang, Y., Han, T., Zhu, Y., Zheng, C.J., Ming, Q.L., Rahman, K., Qin, L.P. (2010) Antidepressant properties of bioactive fractions from the extract of Crocus sativus L. J. Nat. Med. 64(1):24–30. DOI: https://doi.org/10.1007/s11418-009-0360-6.
  48. Yang, L., Xu, H., Hong, Q., Xu, N., Zhang, Y., Tao, R., Li, S., Zhang, Z., Geng, J., Wang, Z., Hu, H., Dong, Y., Chu, Z., Zheng, B., Zhu, J., Geng, M., Gao, Y. (2024) Crocus sativus L. produces anti-inflammatory effects and regulates the NLRP3-NF-κB pathway. Acupunct. Herbal. Med. 4(3):375–385. DOI: https://doi.org/10.1097/HM9.0000000000000088.
  49. Zareena, A.V., Variyar, P.S., Gholap, A.S., Bongirwar, D.R. (2001) Chemi-cal investigation of gamma-irradiated saffron (Crocus sativus L.). J. Agric. Food Chem. 49(2):687–691. DOI: https://doi.org/10.1021/jf000922l.
  50. Zeinali, M., Zirak, M.R., Rezaee, S.A., Karimi, G., Hosseinzadeh, H. (2019) Immunoregulatory and anti-inflammatory properties of Cro-cus sativus (Saffron) and its main active constituents: A review. Iran. J. Basic Med. Sci. 22(4):334–344. DOI: https://doi.org/10.22038/ijbms.2019.34365.8158.
  51. Zwane, B.N., Kamatou, G.P., Viljoen, A.M., Betti, G., Schmidt, M. (2020) Variation in headspace volatiles of saffron determined by GC×GC-ToF-MS. Nat. Pro. Comm. 15(11). DOI: https://doi.org/10.1177/1934578X20967612.