Evaluation of harvest timing and genotypic effect on bioactive composition and antioxidant activity of wheatgrass from traditional and modern wheat cultivars

Abstract

Wheatgrass has gained increasing attention due to its high content of bioactive compounds and associated health benefits. This study aimed to evaluate the effects of harvest timing and wheat variety on plant growth, phytochemical composition—including chlorophyll a (Chl a), chlorophyll b (Chl b), total chlorophyll content (TCC), total phenolic content (TPC), and total flavonoid content (TFC)—as well as antioxidant capacity in wheatgrass obtained from modern bread wheat (Triticum aestivum L.; cv. Tosunbey) and ancestral einkorn wheat (Triticum monococcum ssp. monococcum; cv. IZA). Seedlings were harvested at 10 different growth stages (5 to 15 days old). The results revealed that 9-day-old bread wheatgrass exhibited the highest Chl a (9.43 mg/g FW), Chl b (2.69 mg/g FW), and TCC (12.12 mg/g FW). In contrast, 9-day-old einkorn wheatgrass had the highest TPC (55.18 ± 0.29 mg FAE/g DW) and TFC (263.11 ± 1.92 mg QE/g DW). Notably, the strongest antioxidant activity (2.29 mg/L), comparable to ascorbic acid (2.26 mg/L), was observed in 5-day-old einkorn wheatgrass. The peak levels of TCC, TPC, and TFC were consistently detected on day 9 in both species. These findings highlight the critical role of harvest timing and genetic variation in optimizing the nutritional and functional properties of wheatgrass. While 9-day-old seedlings exhibited the highest phytochemical accumulation, the strongest antioxidant activity was observed in 5-day-old einkorn wheatgrass. Overall, einkorn wheatgrass demonstrated a more potent antioxidant system and higher phenolic and flavonoid content, whereas bread wheatgrass excelled in chlorophyll accumulation, emphasizing distinct compositional advantages between the two wheat varieties.

Keywords

• Antioxidant activity
• Phytochemicals
• Triticum aestivum
• Triticum monococcum ssp. monococcum
• Wheatgrass

References

1. Adhikari, B., Joshi, S., & Maskey, B. (2022). Effects of harvesting period on phytochemicals of wheatgrass (Triticum aestivum, WK 1204 variety). Caraka Tani: Journal of Sustainable Agriculture, 37(1), 36–47. http://dx.doi.org/10.20961/carakatani.v37i1.52076
2. Ahmed, Z., Faisal Manzoor, M., Hussain, A., Hanif, M., & Zia-ud-Din Zeng, X.A. (2021). Study the impact of ultra-sonication and pulsed electric field on the quality of wheat plantlet juice through FTIR and SERS. Ultrasonics Sonochemistry, 76, 105648. https://doi.org/10.1016/j.ultsonch.2021.105648
3. Akbas, E., Kilercioglu, M., Onder, O.N., Koker, A., & Soyler, B., Oztop, M.H. (2017). Wheatgrass juice to wheat grass powder: Encapsulation, physical and chemical characterization. Journal of Functional Foods, 28, 19–27. https://doi.org/10.1016/j.jff.2016.11.010
4. Avisar, A., Cohen, M., Katz, R., Shentzer Kutiel, T., Aharon, A., & Bar-Sela, G. (2020). Wheatgrass juice administration and immune measures during adjuvant chemotherapy in colon cancer patients: Preliminary results. Pharmaceuticals, 13(6), 129. https://doi.org/10.3390/ph13060129
5. Banerjee, S., Katiyar, P., Kumar, V., Waghmode, B., Nathani, S., Krishnan, V., Sircar, D., & Roy, P. (2021). Wheatgrass inhibits the lipopolysaccharide-stimulated inflammatory effect in RAW 264.7 macrophages. Current Research in Toxicology, 2, 116 127. https://doi.org/10.1016/j.crtox.2021.02.005
6. Benincasa, P., Galieni, A., Manetta, A.C., Pace, R., Guiducci, M., Pisante, M., & Stagnari, F. (2015). Phenolic compounds in grains, sprouts and wheatgrass of hulled and nonhulled wheat species. Journal of the Science of Food and Agriculture, 95, 1795–1803. https://doi.org/10.1002/jsfa.6877
7. Benincasa, P., Tosti, G., Farneselli, M., Maranghi, S., Bravi, E., Marconi, O., Falcinelli, B., & Guiducci, M. (2020). Phenolic content and antioxidant activity of einkorn and emmer sprouts and wheatgrass obtained under different radiation wavelengths. Annals of Agricultural Sciences, 65, 68–76. https://doi.org/10.1016/j.aoas.2020.02.001
8. 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, 25–30. https://doi.org/10.1016/S0023-6438(95)80008-5

9. Chang, C.C., Yang, M.H., Wen, H.M., & Chern, J.C. (2002). Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of Food and Drug Analysis, 10(3), 178–182. https://doi.org/10.38212/2224-6614.2748
10. Choi, M.H., Lee, M., Yang, S.H., Shin, H.J. & Jeon, Y. (2021). Hydrophobic fractions of Triticum aestivum L. extracts contain polyphenols and alleviate inflammation by regulating nuclear factor-kappa B. Biotechnology and Bioprocess Engineering, 26, 93–106. https://doi.org/10.1007/s12257-020-0352-7
11. Chauhan, M. (2014). A pilot study on wheat grass juice for its phytochemical, nutritional and therapeutic potential on chronic diseases. International Journal of Chemical Studies, 2(4), 27–34.
12. Dewanto, V., Wu, X., Adom, K.K., & Liu, R.H. (2002). Thermal processing enhances the nutritional value of tomatoes by increasing total antioxidant activity. Journal of Agricultural and Food Chemistry, 50, 3010–3014. https://doi.org/10.1021/jf0115589
13. Eissa, H.A., Mohamed, S.S., & Hussein, A.M.S. (2020). Nutritional value and impact of wheatgrass juice (Green Blood Therapy) on increasing fertility in male albino rats. Bulletin of the National Research Centre, 44(1), 30. https://doi.org/10.1186/s42269-020-0272-x
14. Falcinelli, B., Benincasa, P., Calzuola, I., Gigliarelli, L., Lutts, S., & Marsili, V. (2017). Phenolic content and antioxidant activity in raw and denatured aqueous extracts from sprouts and wheatgrass of einkorn and emmer obtained under salinity. Molecules, 22(12), 2132. https://doi.org/10.3390/molecules22122132
15. Fernandes de Oliveira, A.M., Sousa Pinheiro, L., Souto Pereira, C. K., Neves Matias, W., Albuquerque Gomes, R., Souza Chaves, O., Vanderlei de Souza, M. F., Nobrega de Almeida, R., & Simoes de Assis, T. (2012). Total phenolic content and antioxidant activity of some Malvaceae family species. Antioxidants (Basel), 1, 33–43. https://doi.org/10.3390/antiox1010033
16. Ferruzzi, M.G., & Blakeslee, J. (2007). Digestion, absorption, and cancer preventative activity of dietary chlorophyll derivatives. Nutrition Research, 27(1), 1 12. https://doi.org/10.1016/j.nutres.2006.12.003
17. Garg, M., Sharma, A., Vats, S., Tiwari, V., Kumari, A., Mishra, V., & Krishania, M. (2021). Vitamins in cereals: A critical review of content, health effects, processing losses, bioaccessibility, fortification, and biofortification strategies for their improvement. Frontiers in Nutrition, 8, 254. https://doi.org/10.3389/fnut.2021.586815
18. Gawlik-Dziki, U., Świeca, M., & Dziki, D. (2012). Comparison of phenolic acids profile and antioxidant potential of six varieties of spelt (Triticum spelta L.). Journal of Agricultural and Food Chemistry, 60(18), 4603–4612. https://doi.org/10.1021/jf3011239
19. Ghumman, A., Singh, N., & Kaur, A. (2017). Chemical, nutritional and phenolic composition of wheatgrass and pulse shoots. International Journal of Food Science and Technology, 52(10), 2191–2200. https://doi.org/10.1111/ijfs.13498
20. Hebbani, A.V., Saradamma, B., Kanu, V.R., Malini, A.B., Reddy, V.D., & Chakravarthula, N.V. (2020). Nephroprotective activity of wheatgrass juice against alcohol-induced oxidative damage in rats. Toxicology Mechanisms and Methods, 30(9), 679 685. https://doi.org/10.1080/15376516.2020.1810837
21. Kathuria, D., Hamid, H., Chavan, H.P., Jaiswal, A.K., Thakur, A., & Dhiman, A.K. (2024). A comprehensive review on sprouted seeds bioactives, the impact of novel processing techniques and health benefits. Food Reviews International, 40(1), 370 398. https://doi.org/10.1080/87559129.2023.2169453
22. Kaur, N., Singh, B., Kaur, A., Yadav, M.P., Singh, N., Ahlawat, A.K., & Singh, A.M. (2021). Effect of growing conditions on proximate, mineral, amino acid, phenolic composition and antioxidant properties of wheatgrass from different wheat (Triticum aestivum L.) varieties. Food Chemistry, 341, 128201. https://doi.org/10.1016/j.foodchem.2020.128201
23. Kumar, N.S., Murali, M., Nair, A.M., & Nair, A.S. (2016). Green blood therapy of wheat grass – nature’s finest medicine: A literature review. Journal of Pharmaceutical and Biological Sciences, 11, 57–64. https://doi.org/10.9790/3008-1102045764
24. Li, L., Shewry, P.R., & Ward, J.L. (2008). Phenolic acids in wheat varieties in the HEALTHGRAIN Diversity Screen. Journal of Agricultural and Food Chemistry, 56, 9732 9739. https://doi.org/10.1021/jf801069s
25. Moran, R. (1982). Formulae for determination of chlorophyllous pigments extracted with N,N-dimethylformamide. Plant Physiology, 69, 1376–1381. https://doi.org/10.1104/pp.69.6.1376.
26. Niroula, A., Khatri, S., Timilsina, R., Khadka, D., Khadka, A., & Ojha, P. (2019). Profile of chlorophylls and carotenoids of wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) microgreens. Journal of Food Science and Technology, 56(5), 2758–2763. https://doi.org/10.1007/s13197-019-03768-9
27. Özberk, İ., Atay, S., Cabi, E.E., Özkan, H., & Atlı, A.A. (2016). Türkiye’nin buğday atlası [Wheat Atlas of Turkey]. İstanbul, WWF-Türkiye.
28. Özköse, A., Arslan, D., & Acar, A. (2016). The comparison of the chemical composition, sensory, phenolic and antioxidant properties of juices from different wheatgrass and turfgrass species. Notulae Botanicae Horti Agrobotanici Cluj Napoca, 44(2), 499 507. https://doi.org/10.15835/nbha44210405
29. Qamar, A., Saeed, F., Nadeem, M.T., Hussain, A.I., Khan, M.A., & Niaz, B. (2019). Probing the storage stability and sensorial characteristics of wheat and barley grasses juice. Food Science and Nutrition, 7(2), 554–562. https://doi.org/10.1002/fsn3.841
30. Pandey, K.B., & Rizvi, S.I. (2009). Plant polyphenols as dietary antioxidants in human health and disease. Oxidative Medicine and Cellular Longevity, 2, 897484. https://doi.org/10.4161/oxim.2.5.9498
31. Pehlivan Karakas, F., & Turker, A.U. (2013). An efficient in vitro regeneration system for Bellis perennis L. and comparison of phenolic contents of field-grown and in vitro-grown leaves by LC-MS/MS. Industrial Crops and Products, 48, 162–170. https://doi.org/10.1016/j.indcrop.2013.04.008
32. Pehlivan Karakas, F., & Turker, A.U. (2016). Improvement of shoot proliferation and comparison of secondary metabolites in shoot and callus cultures of Phlomis armeniaca by LC-ESI-MS/MS analysis. In Vitro Cellular and Developmental Biology – Plant, 52, 608 618. https://doi.org/10.1007/s11627-016-9792-3
33. Pehlivan Karakas, F., Keskin, C.N., Agil, F., & Zencirci, N. (2021). Profiles of vitamin B and E in wheat grass and grain of einkorn (Triticum monococcum spp. monococcum), emmer (Triticum dicoccum ssp. dicoccum Schrank.), durum (Triticum durum Desf.), and bread wheat (Triticum aestivum L.) cultivars by LC ESI MS/MS analysis. Journal of Cereal Science, 98, 103177. https://doi.org/10.1016/j.jcs.2021.103177
34. Pehlivan Karakas, F., Keskin, C.N., Agil, F., & Zencirci, N. (2022). Phenolic composition and antioxidant potential in Turkish einkorn, emmer, durum, and bread wheat grain and grass. South African Journal of Botany, 149, 407–415. https://doi.org/10.1016/j.sajb.2022.06.022
35. Reshi, Z.A., Ahmad, W., Lukatkin, A.S., & Javed, S.B. (2023). From nature to lab: A review of secondary metabolite biosynthetic pathways, environmental influences, and in vitro approaches. Metabolites, 13(8), 895. https://doi.org/10.3390/metabo13080895
36. Shamloo, M., Babawale, E.A., Furtado, A., Henry, R.J., Eck, P.K., & Jones, P.J.H. (2017). Effects of genotype and temperature on accumulation of plant secondary metabolites in Canadian and Australian wheat grown under controlled environments. Scientific Reports, 7, 9133. https://doi.org/10.1038/s41598-017-09681-5
37. Sharma, H., Bhandawat, A., Kumar, P., Rahim, M.S., Parveen, A., Kumar, P., Madhawan, A., Rishi, V., & Roy, J. (2020). Development and characterization of bZIP transcription factor based SSRs in wheat. Gene, 756, 144912. https://doi.org/10.1016/j.gene.2020.144912
38. Skoczylas, Ł., Korus, A., Tabaszewska, M., Gędoś, K., & Szczepańska, E. (2018). Evaluation of the quality of fresh and frozen wheatgrass juices depending on the time of grass harvest. Journal of Food Processing and Preservation, 42, e13401. https://doi.org/10.1111/jfpp.13401
39. Szablińska-Piernik, J. & Lahuta, Ł. B. (2023). Changes in polar metabolites during seed germination and early seedling development of pea, cucumber, and wheat. Agriculture, 13(12), 2278. https://doi.org/10.3390/agriculture13122278
40. Thakur, N., Dhaliwal, H.S., & Sharma, V. (2019). Chemical composition, minerals and vitamins analysis of lyophilized wheatgrass juice powder. International Journal on Emerging Technologies, 10(4), 137–144.
41. Thakur, N., Dhaliwal, H., & Sharma, V. (2020). Qualitative and quantitative RP-HPLC-PDA method of analysis of polyphenols in lyophilized wheat seedling juice powder. International Journal on Emerging Technologies, 11(2), 36–43.
42. Tekin, M., Cengiz, M.F., Abbasov, M., Aksoy, A., Canci, H., & Akar, T. (2018). Comparison of some mineral nutrients and vitamins in advanced hulled wheat lines. Cereal Chemistry, 95, 436–444. https://doi.org/10.1002/cche.10045
43. Tullo, A., & Abera, S. (2023). Review on nutrient contents and health benefits of wheatgrass juice. International Journal of Smart Agriculture, 1, 32–39. https://doi.org/10.54536/ijsa.v1i1.2193
44. Verma, B., Hucl, P., & Chibbar, R.N. (2009). Phenolic acid composition and antioxidant capacity of acid and alkali hydrolysed wheat bran fractions. Food Chemistry, 116, 947 954. https://doi.org/10.1016/j.foodchem.2009.03.060
45. Wang, L., Yao, Y., He, Z., Wang, D., Liu, A., & Zhang, Y. (2013). Determination of phenolic acid concentrations in wheat flours produced at different extraction rates. Journal of Cereal Science, 57, 67–72. https://doi.org/10.1016/j.jcs.2012.09.013
46. Wangcharoen, W., & Phimphilai, S. (2016). Chlorophyll and total phenolic contents, antioxidant activities and consumer acceptance test of processed grass drinks. Journal of Food Science and Technology, 53(12), 4135–4140. https://doi.org/10.1007/s13197-016-2380-z
47. Ward, J.L., Poutanen, K., Gebruers, K., Piironen, V., Lampi, A.M., Nyström, L., Andersson, A.A.M., Åman, P., Boros, D., Rakszegi, M., Bedő, Z., & Shewry, P.R. (2008). The HEALTHGRAIN cereal diversity screen: Concept, results, and prospects. Journal of Agricultural and Food Chemistry, 56, 9699–9709. https://doi.org/10.1021/jf8009574
48. Watanabe, N. (2017). Breeding opportunities for early, free-threshing and semi-dwarf Triticum monococcum L. Euphytica, 213, 201. https://doi.org/10.1007/s10681-017-1987-0
49. Zhang, Y.M., Guo, P., Xia, X., Guo, H., & Li, Z. (2021). Multiple layers of regulation on leaf senescence: New advances and perspectives. Frontiers in Plant Science, 12, 788996. https://doi.org/10.3389/fpls.2021.788996
50. Zielińska-Dawidziak, M. (2021). Influence of stress conditions on the quality of obtained sprouts. Quality Assurance and Safety of Crops & Foods, 13(2), 1 12. https://doi.org/10.15586/qas.v13i2.836