Document Type : Full Research Paper

Authors

1 MSc. student, Department of Food Science and Technology, Faculty of Animal Science and Food Technology, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Iran.

2 Professor, Department of Food Science and Technology, Faculty of Animal Science and Food Technology, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Iran

3 Department of Food Science and Technology, Faculty of Animal Science and Food Technology, Agricultural Sciences and Natural Resources University of Khuzestan, Mollasani, Iran

Abstract

Introduction: Shirazi thyme is a perennial herbaceous plant related to Lamiaceae family growing in central and southern regions of Iran. This study aimed to assess chemical composition, bioactive functional groups, antioxidant potential, total phenol, and total flavonoids of Shirazi thyme essential oil (STEO) and also to evaluate its cell toxicity effect against the colorectal cancer cell line (HT29).
 
Materials and Methods: The chemical composition of STEO was identified by gas chromatography-mass spectrometry. The bioactive functional groups of STEO were measured by Fourier transform infrared at spectra range of 500- 4000 cm-1. The total phenol and total flavonoids of STEO were determined by folin- ciocalteu and aluminum chloride colorimetric method, respectively. MMT method was performed to measure the cell toxicity of STEO against the colorectal cancer cell line (HT29). The cells were cultured on DMEM high glucose medium supplemented with 10% fetal bovine serum and penicillin/streptomycin and incubated at 37°C with 95% relative humidity and 5% CO2 concentration. The antioxidant activity of STEO was evaluated by DPPH and ABTS free radicals scavenging assays. Duncan test at 5% probability and SPSS software (version 18) was performed to compare the means of obtained results.
 
Results and Discussion: Based on GC/MS spectrometry, a total of 29 constituents were identified and quantified in the STEO representing more than 99% of total constituents. Thymol with 39.3% and Carvacrol with 30% were the most constituents of STEO. The other major compound of STEO were Benzene (8.52%), γ-Terpinene (5.27%), and Caryophyllene (2.97%). The presence of peak at a wavelength of 320- 1000 cm-1 demonstrates O-C bound that could be related to organic compounds such as alcohols, carboxylic acids, esters, and ethers. The peaks occurred at spectral range between 2800- 3000 cm-1 (particularly at 2869.6 up to 2960.8 cm-1) are related to the stretching mode of C-H bounds and are mostly associated with alcoholic compounds in the essence. The amount of total phenol and flavonoids of STEO were 64.05 mg gallic acid equivalent/g and 11.68 mg quercetin equivalent/g, respectively. Results obtained from antioxidant activity of STEO with DPPH and ABTS free radicals scavenging showed that by increasing of STEO, the inhibition level of free radicals was enhanced. The inhibition percent of free radicals by using DPPH and ABTS at 1000 ppm concentration were 63.69% and 64.33%, respectively. The cell toxicity results of different STEO concentrations against the cell line HT29 revealed that survival of HT29 cells at 1, 3.125, 6.25, 12.5, 25, 50, 100 and 200 ppm of STEO were 100, 70.15, 61.19, 59.33, 38.83, 25.68, 20.65 and 12.2%. According to the results of this study, as the concentration of STEO increased, its effect on the cell line HT29 enhanced and the cell percentage viability decreased. Some antioxidant activity of STEO could be interrelated to its phenolic compounds. Based on the results of antioxidant activity, the amount of total phenol and flavonoids and the effect of STEO on cell toxicity against the cell line HT29, it seems that the use of STEO is applicable in the pharmaceutical and food industries

Keywords

  1. Abdalan, S., Baghbani-Arani, F. & Sadat Shandiz, S. A., (2018). Evaluation of Anticancer Effect of Aqueous and Hydroalcoholic Extracts of Quercusin Fectoria Leaf against Colon Cancer HT29 Cell Line. Journal of Arak University Medical Sciences, 21(4), 48-57. (In Persian)
  2. Abdelhady MI, Aly HAH., 2012. Antioxidant antimicrobial activities of Callistemon comboynensis essential oil. Free Radical Antioxid, 2(1), 37-41. https://doi.org/10.5530/ax.2012.2.8
  3. Aljabeili, H. S., Barakat, H. & Abdel-Rahman, H. A., (2018). Chemical composition, antibacterial and antioxidant activities of Thyme essential oil (Thymus vulgaris). Food and Nutrition Sciences, 9(05), 433-446.‏doi: 4236/fns.2018.95034
  4. Ahmadi, E., Abdollahi, A., Najafipour, S., Meshkibaf, M.H., Fasihi Ramandi, M., Namdar, N., Abdollahi Kheyr Abadi, S., Mousavi, S. M., Samizadeh, B. & Allahverdi, G., (2016). Surveying the Effect of the Phenol Compounds on Antibacterial Activity of Herbal Extracts: In vitro Assessment of Herbal Extracts in Fasa-Fars Province. Journal of Fasa University of Medical Sciences, 6(2), 210-220. (In Persian)
  5. Aliakbarlu, J., Sadaghiani, S. K. & Mohammadi, S., (2013). Comparative evaluation of antioxidant and antifood-borne bacterial activities of essential oils from some spices commonly consumed in Iran. Food Science and Biotechnology, 22(6), 1487-1493.‏ https://doi.org/10.1007/s10068-013-0242-2
  6. Alizadeh Behbahani, B., Shahidi, F.,Tabatabaei Yazdi, F., Mortazavi, S.A. & Mohebbi, M., (2017). Use of Plantago major seed mucilage as a novel edible coating incorporated with Anethum graveolens essential oil on shelf life extension of beef in refrigerated storage. International Journal of Biological Macromolecules, 94, 515-526.‏ https://doi.org/10.1016/j.ijbiomac.2016.10.055
  7. Alizadeh Behbahani, B., Noshad, M. & Falah, F., (2019). Study of chemical structure, antimicrobial, cytotoxic and mechanism of action of Syzygium aromaticum essential oil on foodborne pathogens. Potravinarstvo Slovak Journal of Food Sciences, 13(1), 875-883.‏
  8. Alsaraf, S., Hadi, Z., Al-Lawati, W. M., Al Lawati, A. A. & Khan, S. A., (2020). Chemical composition, in vitro antibacterial and antioxidant potential of Omani Thyme essential oil along with in silico studies of its major constituent. Journal of King Saud University-Science, 32(1),1-21. https://doi.org/10.1016/j.jksus.2019.09.006
  9. Amin, M., Kalantar, E., Mohammad-Saeid, N. & Ahsan, B., Antibacterial effect and physicochemical properties of essential oil of Zataria multiflora Boiss. Asian Pacific Journal of Tropical Medicine, 3(6), 439-442.https://doi.org/10.1016/S1995-7645(10)60105-8
  10. Avaei, A., Mohamadi Sani, A. & Mahmoodzadeh Vaziri, B., (2015). Chemical composition and antimicrobial effect of the essential oil of Zataria multiflora Boiss endemic in Khorasan-Iran. Asian Pacific Journal of Tropical Disease, 5(3), 181-185. https://doi.org/10.1016/S2222-1808(14)60649-6
  11. Barzegar, H., Alizadeh Behbahani, B. & Mehrnia, M. A., (2020). Quality retention and shelf life extension of fresh beef using Lepidium sativum seed mucilage-based edible coating containing Heracleum lasiopetalum essential oil: an experimental and modeling study. Food Science and Biotechnology, 29(5), 717-728.‏ https://doi.org/10.1007/s10068-019-00715-4
  12. Blowman, K., Magalhães, M., Lemos, M., Cabral, C., Pires, I. (2018). Anticancer properties of essential oils and other natural products. Evidence-Based Complementary and Alternative Medicine, 2018, 3149362. https://doi.org/10.1155/2018/3149362
  13. Brand-Williams, W., Cuvelier, M. E. & Berset, C., (1995). Use of a free radical method to evaluate antioxidant activity. LWT-Food Science and Technology, 28(1), 25-30.‏ https://doi.org/10.1016/S0023-6438(95)80008-5
  14. Brodowska, K. M., (2017). Natural flavonoids: classification, potential role, and application of flavonoid analogues. European Journal of Biological Research, 7(2), 108-123.‏
  15. Cui, W., Phillips, G. O., Blackwell, B. & Nikiforuk, J., (2007). Characterisation and properties of Acacia senegal (L.) Willd. var. senegal with enhanced properties (Acacia (sen) Supergum (TM)): Part 4. Spectroscopic characterisation of Acacia senegal var. senegal and Acacia (sen) Supergum (TM) arabic. Food Hydrocolloids, 21(3): 347–352.
  16. Dabbagh Moghaddam, A., Kazemi, M., Movaffagh, J. & Sharifan, A., (2019). Design of Zein Electrospinning Nanofiber Packaging Containing Zataria Multiflora Essential Oil to Preserve the Ration Food. Journal of Food Technology and Nutrition, 16(3), 91-102. (In Persian)
  17. Dalali Isfahani, L., Monajemi, R. & Amjad, L., (2013). Cytotoxic Effects of Extract and Essential oil Leaves of Achillea wilhelmsiiKoch on Colon Cancers Cells. Experimental Animal Biology, 1(3), 1-6. (In Persian)
  18. El Abed, N., Kaabi, B., Smaali, M. I., Chabbouh, M., Habibi, K., Mejri, M., Marzouki, M. N. & Ben Hadj Ahmed, S., (2014). Chemical composition, antioxidant and antimicrobial activities of Thymus capitata essential oil with its preservative effect against Listeria monocytogenes inoculated in minced beef meat. Evidence-Based Complementary and Alternative Medicine, 2014.‏ https://doi.org/10.1155/2014/152487
  19. Fazli, R., Nazarnezhad, N. & Ebrahimzadeh, M., (2013). Evaluation of the Antioxidant Capacities and Total Phenolic Contents of Beech, Hornbeam and Poplar Barks. Journal of Forest and Wood Products, 66(3), 339-349. (In Persian)
  20. Geraminezhad, F. & Roostaei-Ali Mehr, M., 2018. Effect of Zataria multiflora essential oil on rooster semen during storage at 4 C. Iranian Journal of Veterinary Science and Technology, 10(1), 21-26.‏
  21. Golkar, P., Mosavat, N. & Jalali, S. A. H., (2020). Essential oils, chemical constituents, antioxidant, antibacterial and in vitro cytotoxic activity of different Thymus species and Zataria multiflora collected from Iran.South African Journal of Botany, 130, 250-258.‏ https://doi.org/10.1016/j.sajb.2019.12.005
  22. Hossain, M. A. & Rahman, S. M. M., (2011). Total phenolics, flavonoids and antioxidant activity of tropical fruit pineapple. Food Research International, 44(3), 672-676.‏ https://doi.org/10.1016/j.foodres.2010.11.036
  23. Huyut, Z., Beydemir, Ş. & Gülçin, İ., (2017). Antioxidant and antiradical properties of selected flavonoids and phenolic compounds.Biochemistry Research International, .‏ https://doi.org/10.1155/2017/7616791
  24. Janitermi, M., Nemati, F. & Mohammadpour, G., (2015). Cytotoxic effect of Zataria multiflora on gastric cancer cell line (AGS) and normal fibroblast cells. Cumhuriyet Üniversitesi Fen-Edebiyat Fakültesi Fen Bilimleri Dergisi, 36(3), 1876-1884.‏
  25. Janitermi, M. & Nemati, F., (2015). Cytotoxic effect of Zataria multiflora on breast cancer cell line (MCF-7) and normal fibroblast cells. Cumhuriyet Üniversitesi Fen-Edebiyat Fakültesi Fen Bilimleri Dergisi, 36(3), 1895-1904.
  26. ‏Karaman, Ş., Tütem, E., Başkan, K.S. & Apak, R., (2010). Comparison of total antioxidant capacity and phenolic composition of some apple juices with combined HPLC–CUPRAC assay. Journal of Food Chemistry, 120(4), 1201-1209.‏ https://doi.org/10.1016/j.foodchem.2009.11.065
  27. Jooyandeh, H. & Haji Ebrahimi Forushani, Z., (2019). Extraction of polysaccharide from Pleurotus ostreatus by ultrasound and evaluation of its antioxidant activity. Journal of Food Science and Technology, 16(89), 85-100. (In Persian)
  28. Kavoosi, G. & Rabiei, F., (2015). Zataria multiflora: chemical and biological diversity in the essential oil. Journal of Essential Oil Research, 27(5), 428-436. https://doi.org/10.1080/10412905.2015.1031917
  29. Kavoosi, G., Purfard, A. M. & Aram, F., 2012. Radical scavenging properties of essential oils from Zataria multiflora and Ferula assafoetida. Asian Pacific Journal of Tropical Biomedicine, 2(3), S1351-S1356.‏ https://doi.org/10.1016/S2221-1691(12)60415-8
  30. Khakzad, S., Rahmani, F., Hojjati, M. & Tabandeh, M.R., (2019). Anti-carcinogenic effects of Satureja khuzistanica and Zataria multiflora essential oils on K562 cell line proliferation. Journal of Food and Bioprocess Engineering, 2(2), 127-132.
  31. Khazraei-Moradian, S., Andalib, A., Ganjalikhani-Hakemi, M., Safari, Z., Zare, A. & Kardar, G., (2014). The Effect of Protein Extract of Licorice Root in proliferation of HT-29 and CT26 Cancer Cell Lines. Journal of Isfahan Medical School, 32(298), 1-9. (In Persian)
  32. Khoshsokhan, F., Babalar, M., Poormeidani, A. & Fatahi, M. R., (2015). Antioxidant Activity, Total Phenolics and Oil Content of Some Thymus kotschyanus and Thymus daenensis Plant Production Technology, 15(1), 153-162. (In Persian)
  33. Mahboubi, M., Heidarytabar, R., Mahdizadeh, E. & Hosseini,, (2017). Antimicrobial activity and chemical composition of Thymus species and Zataria multiflora essential oils. Agriculture and Natural Resources, 51(5), 1-18. https://doi.org/10.1016/j.anres.2018.02.001
  34. Mantashlo, J., Deljoo, A. & Piri, Kh. (2017). The Study of Flavonoids and Antioxidant Power of Etanolic, Metanolic, Hydroalcholic and Etylacetatic Extracts of Branch and Stem Bark of Salix Alba. Journal of Molecular and Cellular Research (Iranian Journal of Biology), 30(3), 381-391. (In Persian)
  35. Mehran, M., Hosseini, H., Hatami, A., Taghizade, M. & Safaei, A., (2016). Investigation of seven species of essential oils of thyme and comparison their antioxidant properties. Journal of Medicinal Plants, 15(58), 134-140. (In Persian)
  36. Mehrnia, M. A., Dehghan, N. & heidari, M., (2019). Effect of drying method and solvent type on antioxidant properties and chemical composition of sacred fig (Ficus religiosa). Journal of Food Science and Technology, 16(95), 27-37. (In Persian)
  37. Marinova, D., Ribarova, F. & Atanassova, M., (2005). Total phenolics and total flavonoids in Bulgarian fruits and vegetables. Journal of the University of Chemical Technology and Metallurgy, 40(3), 255-260.‏
  38. Mirzaee, A., Jaberi Hafashani, H. & Madani, A., (2012). Antioxidant activities, total phenols and total Flavonoids assay of Origanurm vulgare, Teucrium polium and Thymus daensis. Hormozgan Medical Journal, 15(4), 285-294. (In Persian)
  39. Mohammed, K. A. K., Abdulkadhim, H. M. & Noori, S. I., (2016). Chemical composition and anti-bacterial effects of clove (Syzygium aromaticum) flowers. International Journal of Current Microbiology and Applied Sciences, 5(2), 483-489.
  40. Mojaddar Langroodi, A., Tajik, H. & Mehdizadeh, T., (2019). Antibacterial and antioxidant characteristics of Zataria multiflora Boiss essential Oil and hydroalcoholic extract of Rhus coriaria Journal of Food Quality and Hazards Control, 6(1), 16-24.
  41. Mortazaei, S., Mahmoudvand, H. & Rafieian, R., 2016. Phytochemical properties of some Iranian medicinal plants. Journal of Chemical and Pharmaceutical Sciences, 9(4), 3427-3432.‏
  42. Moshafi, M. H., Mansouri, Sh., Sharififar, F. & Khoshnoodi, M., (2007). Antibacterial and Antioxidant Effects of the Essential Oil and Extract of Zataria Multiflora Journal of Kerman University of Medical Sciences, 14(1), 33-43. (In Persian)
  43. Najafpour Navaei, M. & Mirza, M., (2015). Essential Oil Composition of Zataria Multiflora In Different Province of Iran. Eco-phytochemical Journal of Medical Plants, 2(4), 43-49. (In Persian)
  44. Nasiri, E., Moosavi-Nasab, M., Shekarforoush, S. S. & Golmakani, M. T., (2014). The effects of Zataria multiflora on inhibition of polyphenoloxidase and melanosis formation in shrimp (Litopenaeus vannamei). Iranian Scientific Fisheries Journal, 23(3), 109-118. (In Persian)
  45. Rahimifard, N., Pakzad, S. R., Shoeibi, Sh., Hedayati, M. H., Hajimehdipour, H., Motaharinia, V., Mehrafshan, L., Javadi A. & Pirali Hamedani, M., (2009). Effects of Essential oil and Extract of Thymus vulgaris, Zataria multiflora and Eugenia caryophilata on Vero, Hela, HepII cell lines by MTT Assay. Journal of Medicinal Plants, 8(30), 152-156. (In Persian)
  46. Ramezanian, A., Azadi, M., Mostowfizadeh-Ghalamfarsa, R. & Saharkhiz, M.J., (2016). Effect of Zataria multiflora​ Boiss and Thymus vulgaris​ essential oils on black rot of ‘Washington Navel’orange fruit. Postharvest Biology and Technology, 112, 152-158.‏ https://doi.org/10.1016/j.postharvbio.2015.10.011
  47. Rezzoug, M., Bakchiche, B., Gherib, A., Roberta, A., Guido, F., Kilincarslan, O., Mammadov, R. & Bardaweel, S. K., (2019). Chemical composition and bioactivity of essential oils and Ethanolic extracts of Ocimum basilicum and Thymus algeriensis Boiss. & Reut. from the Algerian Saharan Atlas. ‏BMC Complementary and Alternative Medicine, 19(1), 146.‏ https://doi.org/10.1186/s12906-019-2556-y
  48. Sadeghi, H., Robati, Z. & Saharkhiz, M. J., (2015). Variability in Zataria multiflora Bioss. essential oil of twelve populations from Fars province, Iran. Industrial Crops and Products, 67, 221-226. https://doi.org/10.1016/j.indcrop.2015.01.021
  49. Salhe Abadi, S. & Mehraban Sang Atash, M., (2015). Evaluation of the antioxidant activity and total phenols, flavonoids in methanolic, dichloromethane and ethyl acetate extracts of aerial parts of Rubia florida. Journal of North Khorasan University of Medical Sciences, 7(1), 101-112. (In Persian)
  50. Seyedalipour, B., Hasani, A., Ebrahimzadeh, M. A. & Taravati, A., (2016). Antioxidant activity, total flavonoid and total phenolic contents of extracts taken from aerial parts of Ballota platyloma using three different methods: percolation, ultrasonic and polyphenolic fraction. Journal of Kashan University of Medical Sciences, 20(2), 147-156.‏
  51. Shahsavari, N., Barzegar, M., Sahari, M. A. & Naghdi Badi, H., (2008). An Investigation on the Antioxidant Activity of Essential Oil of Zataria multiflora in Soy Bean Oil. Journal of Medicinal Plants, 7(28), 56-68. (In Persian)
  52. Sharafati Chaleshtori, R., Rafieian Kopaei, M., Rokni, N., Mortezaei, S. & Sharafati Chaleshtori, A., (2013). Antioxidant Activity of Zataria Multiflora Hydroalcoholic Extract and Its Antibacterial Effect on Staphylococcus Aureus. Journal of Mazandaran University of Medical Sciences, 22(1), 88-94. (In Persian)
  53. Sharififar, F., Miri-Moghaddam, M., Nematollahi-Mahani, S. N., Forootanfar, H. & Asgharian Rezee, M., (2017). Cytotoxicity effect of Zataria multiflora on two human colon carcinoma cell lines. Research Journal of Pharmacognosy, 4(4), 65-70.‏
  54. Shokrzadeh, M., Azadbakht, M., Ahangar, N., Naderi, H. & Saeedi Saravi, S. S., (2010). Comparison of the cytotoxic effects of Juniperus sabina and Zataria multiflora extracts with Taxus baccata extract and Cisplatin on normal and cancer cell lines.Pharmacognosy Magazine, 6(22), 102-105.‏ DOI: 4103/0973-1296.62894
  55. Symonowicz, M. & Kolanek, M., (2012). Flavonoids and their properties to form chelate complexes.‏ Biotechnology and Food Sciences, 76(1), 35-41.
  56. Yonesi, B., Mirzaie, A. & Aliasgari, E., (2017). Cytotoxicity and apoptotic effect of oxaliplatin on colon cancer cell line (HT29) and analysis of caspase 3 and caspase 9 gene expression using Real Time PCR method. Journal of Cell and Tissue, 8(4), 364-373. (In Persian)
  57. Zangiabadi, M., Sahari, M. A., Barzegar, M. & Naghdi Badi, H., (2012). Zataria multiflora and Bunium persicum essential oils as two natural antioxidants. Journal of Medicinal Plants, 11(41), 8-21.‏
  58. Ziaee, E., Razmjooei, M., Shad, E. & Eskandari, M.H., (2018). Antibacterial mechanisms of Zataria multiflora essential oil against Lactobacillus curvatus. LWT-Food Science and Technology, 87, 406-412. https://doi.org/10.1016/j.lwt.2017.08.089

 

CAPTCHA Image