Insight into possible therapeutic applications of Ephedra transitoria: in-vitro biological, toxicological activities and GC-MS analysis of aerial parts’ extract


  • Kareem M. YOUNES University of Ha’il, College of Pharmacy, Department of Pharmaceutical Chemistry, 81442 Hail; Cairo University, Faculty of Pharmacy, Department of Analytical Chemistry, 11562 Cairo (SA)
  • Fawaz ALHEIBSHY University of Ha’il, College of Pharmacy, Department of Pharmaceutics, 81442 Hail; Aden University, Faculty of Pharmacy, Department of Pharmaceutics, 6075 Aden (SA)
  • Amal M. ALRASHIEDI University of Ha’il, College of Pharmacy, 81442 Hail (SA)
  • Lamya A. ALWHADANI University of Ha’il, College of Pharmacy, 81442 Hail (SA)
  • Shomokh M. ALSHAMMARI University of Ha’il, College of Pharmacy, 81442 Hail (SA)
  • Asmaa S. ALSHAMMARI University of Ha’il, College of Pharmacy, 81442 Hail (SA)
  • Manawy K. ALBALAWI University of Ha’il, College of Pharmacy, 81442 Hail (SA)
  • Turki A. ALMUTIRI University of Ha’il, College of Pharmacy, 81442 Hail (SA)
  • Ghazi ALJOHANI University of Ha’il, College of Pharmacy, 81442 Hail (SA)



antibacterial, antioxidant, apoptosis, cytotoxic, Ephedra


Most Arab nations have arid regions where Ephedra transitoria, a plant, flourishes. It has a variety of medical benefits. It belongs to the Ephedraceae family. The plant's aerial portions were procured from the Hail region of Saudi Arabia, methanol was used to extract bioactive phytochemicals with variable solubility in an efficient manner, and the resulting extract was tested on several cancer cell lines. The extract's cytotoxic potential was measured by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay method against standard vinblastine sulfate and for each cell line, the pertinent half maximal inhibitory concentration values were calculated. Additional apoptosis assays were conducted. Also, the antioxidant activity of plant extract was examined using 2, 2-Diphenyl-1-picrylhydrazyl and [2, 2’-azinobis-(3-ethylbenzothiazoline-6-sulfonate)] methods compared to ascorbic acid standard. Also, antimicrobial effect of extract was evaluated on different gram positive and gram-negative bacteria by zone inhibition and minimum inhibitory concentration assay methods. GC-MS analysis was further applied to identify major bioactive substances. Extract showed good cytotoxic activity against both lung and liver cancer cells with relevant half maximal inhibitory concentration values comparable to those of standard. Extract resulted in S-phase arrest, primarily via inducing apoptosis mediated through activating caspase-3, Bax, and p53 proteins. Plant extract showed good antibacterial activities against S. aureus, E. coli and B. subtilis. Also, it showed high antioxidant activity with relevant half maximal inhibitory concentration values comparable to those of standard. Some bioactive compounds were separated and identified by GC-MS analysis such as; ephedrine and quercetin. ephedra transitoria extract possess high cytotoxic activity against lung and liver cancer cells via S-phase arrest mediated by activation of apoptotic proteins. It also possesses high antioxidant and antibacterial activities.


Abo-Ashour MF, Eldehna WM, George RF, Abdel-Aziz MM, Elaasser MM, Gawad NMA, … Abou-Seri SM (2018). Novel indole-thiazolidinone conjugates: Design, synthesis and whole-cell phenotypic evaluation as a novel class of antimicrobial agents. European Journal of Medicinal Chemistry 160:49-60.

Ahmeda A, Zangeneh A, Zangeneh MM (2020). Green synthesis and chemical characterization of gold nanoparticle synthesized using Camellia sinensis leaf aqueous extract for the treatment of acute myeloid leukemia in comparison to daunorubicin in a leukemic mouse model. Applied Organometallic Chemistry 34:e5290.

Al-Khalil S, Alkofahi A, El-Elsawi D, Al-Shibib A (1998). Transitorine, a new Quinoline alkaloid from Ephedra transitoria. Journal of Natural Products 61(2):262-263.

Bashyal BP, Wijeratne EK, Faeth SH, Gunatilaka AL, Globosumones AC (2005). Cytotoxic orsellinic acid esters from the Sonoran Desert endophytic fungus Chaetomium globosum. Journal of Natural Products 68(5):724-728.

Calzada F, Bautista E (2020). Plants used for the treatment of diarrhea from Mexican flora with amoebicidal and giadicidal activity, and their phytochemical constituents. Journal of Ethnopharmacology 253:112676.

Cano A, Maestre AB, Hernández-Ruiz J, Arnao MB (2023). ABTS/TAC methodology: main milestones and recent applications. Processes 11:185.

Chien JH, Chang KF, Lee SC, Lee CJ, Chen YT, Lai HC, ... Tsai NM (2022). Cedrol restricts the growth of colorectal cancer in vitro and in vivo by inducing cell cycle arrest and caspase-dependent apoptotic cell death. International Journal of Medical Sciences 19(13):1953.

Cruz-Ramirez SG, Lopez-Saiz CM, Rosas-Burgos EC, Cinco-Moroyoqui FJ, Velazquez C, Hernandez J (2021). Antimutagenic bis (2-ethylhexyl) phthalate isolated from octopus (Paraoctopus vulgaris). Food Science and Technologies 41(2).

David AV, Arulmoli R, Parasuraman S (2016). Overview of biological importance of quercetin: A bioactive flavonoid. Pharmacognosy Reviews 10(20):84-89.

Eldehna WM, Abo-Ashour MF, Ibrahim HS, Al-Ansary GH, Ghabbour HA, Elaasser MM, Safwat NA (2018). Novel [(3-indolylmethylene) hydrazono]indolin-2-ones as apoptotic anti-proliferative agents: design, synthesis and in vitro biological evaluation. Journal of Enzyme Inhibition and Medicinal Chemistry 33:686-700

González-Juárez DE, Escobedo-Moratilla A, Flores J, Hidalgo-Figueroa S, Martínez-Tagüeña N, Morales-Jiménez J, … Trujillo J (2020). A review of the Ephedra genus: distribution, ecology, ethnobotany, phyto-chemistry and pharmacological properties. Molecules 25(14):3283.

Guo JP, Pang J, Wang XW, Shen ZQ, Jin M, Li JW. (2006). In vitro screening of traditionally used medicinal plants in China against enteroviruses. World Journal of Gastroenterology: WJG 12(25):4078.

Hikino H, Ogata K, Konno CH, Sato S. (1983). Hypotensive actions of Ephedradines, Macrocyclic spermine alkaloids of Ephedra roots. Planta Medica 48(8):290-293. 1055/s-2007-969936

Hollander JL, Vander Wall SB, Baguley JG (2010). Evolution of seed dispersal in North American Ephedra. Evolutionary Ecology 24:333-345.

Oh I, Yang WY, Park J, Lee S, Mar W, Oh KB, Shin J (2011). In vitro Na+/K+-ATPase inhibitory activity and antimicrobial activity of sesquiterpenes isolated from Thujopsis dolabrata. Archives of Pharmacal Research 34:2141-2147.

Kalayou S, Haileselassie M, Gebre-Egziabher G, Tiku'e T, Sahle S, Taddele H, Ghezu M (2012). In–vitro antimicrobial activity screening of some ethnoveterinary medicinal plants traditionally used against mastitis, wound and gastrointestinal tract complication in Tigray Region, Ethiopia. Asian Pacific Journal of Tropical Biomedicine 2:516-522.

Khan Z, Nath N, Rauf A, Emran TB, Mitra S, Islam F, ... Thiruvengadam M (2022). Multifunctional roles and pharmacological potential of β-sitosterol: Emerging evidence toward clinical applications. Chemico-Biological Interactions 110117.

Khazaei S, Abdul Hamid R, Ramachandran V, Mohd Esa N, Pandurangan AK, Danazadeh F, Ismail P (2017). Cytotoxicity and proapoptotic effects of Allium atroviolaceum flower extract by modulating cell cycle arrest and caspase-dependent and p53-independent pathway in breast cancer cell lines. Evidence-Based Complementary and Alternative Medicine.

Kittana N, Abu-Rass H, Sabra R, Manasra L, Hanany H, Jaradat N, Hussein F, Zaid AN (2017). Topical aqueous extract of Ephedra alata can improve wound healing in an animal model. Chinese Journal of Traumatology 20(02):108-113.

Kyslychenko V, Karpiuk U, Diakonova I, Mohammad Abu-Darwish (2010). Phenolic compounds and terpenes in the green parts of glycine Hispida. Advances in Environmental Biology 4(3):490-494.

Latif A, Amer HM, Hamad ME, Alarifi SAR, Almajhdi FN (2014). Medicinal plants from Saudi Arabia and Indonesia: In vitro cytotoxicity evaluation on Vero and Hep-2 cells. Journal of Medicinal Plants Research 8:1065-1073.

Liang S, Meng X, Wang Z, Liu J, Kuang H, Wang Q (2018). Polysaccharide from Ephedra sinica inhibits inflammation expression by regulating Factor-β1/Smad2 signaling. International Journal of Biological Macromolecules 106:947-954.

Ling LT, Yap S-A, Radhakrishnan AK, Subramaniam T, Cheng HM, Palanisamy UD (2009). Standardized Mangifera indica extract is an ideal antioxidant. Food Chemistry 113:1154-1159.

Meduru H, Wang YT, Tsai JJ, Chen YC (2016). Finding a potential dipeptidyl peptidase-4 (DPP-4) inhibitor for type-2 diabetes treatment based on molecular docking, pharmacophore generation, and molecular dynamics simulation. International Journal of Molecular Sciences 17(6):920.

Mendelovich M, Shoshan M, Fridlender M, Mazuz M, Namder D, Nallathambi R, Selvaraj G, Kumari P, Ion A, Wininger S, Nasser A (2017). Effect of Ephedra foeminea active compounds on cell viability and actin structures in cancer cell lines. Journal of Medicinal Plants Research 11(43):690-702.

Miller NJ, Rice-Evans C, Davies MJ, Gopinathan V, Milner A (1993). A novel method for measuring antioxidant capacity and its application to monitoring the antioxidant status in premature neonates. Clinical Science 84:407-412. https://

Naeim H, El-Haweit A, Abdel Rahman RA, Hussein A, El-Demellawy MA, Embaby AM (2020). Antibacterial activity of Centaurea pumilio L. root and aerial part extract against some multidrug resistant bacteria. BMC Complementary Medicine and Therapies 20:79.

Parveen S, Shabbir A, Butt AM, Imran M, Jamil A, Asim A, Mashaal K (2023). Amelioration of rheumatoid arthritis in rats by 4-allyanisole through modulation of inflammatory mediator. Research Square.

Pisani C, Ramella M, Boldorini R, Loi G, Billia M, Boccafoschi F, Volpe A, Krengli M (2020). Apoptotic and predictive factors by Bax, Caspases 3/9, Bcl-2, p53 and Ki-67 in prostate cancer after 12 Gy single-dose. Scientific Reports 10:1-10.

Prakash M, Raj B, Murthy K, Mohite M (2018). Biological functions of epicatechin: plant cell to human cell health. Journal of Functional Foods 52:14-24.

Rajendrasozhan S, El Moll H, Snoussi M, Romeilah RM, Shalaby EA, Younes KM, El-Beltagi HS (2021). Phytochemical screening and antimicrobial activity of various extracts of aerial parts of Rhanterium epapposum. Processes 9:1351.

Ramadan MA, Shawkey AE, Rabeh MA, Abdellatif AO (2019). Expression of P53, BAX, and BCL-2 in human malignant melanoma and squamous cell carcinoma cells after tea tree oil treatment in vitro. Cyto-technology 71:461-488.

Sajeesh T, Parimelazahagan (2014). Analgesic, anti-inflammatory, and GC-MS Studies on Castanospermum australe A. Cunn. & C. Fraser ex Hook. The Scientific World Journal 587807.

Salam HS, Tawfik MM, Elnagar MR, Mohammed HA, Zarka MA, Awad NS (2022). Potential apoptotic activities of Hylocereus undatus peel and pulp extracts in MCF-7 and Caco-2 cancer cell lines. Plants 11:2192.

Samadi N, Manayi A, Vazirian M, Samadi M, Zeinalzadeh Z, Saghari Z, Abadian N, Mozaffarian VOA, Khanavi M (2012). Chemical composition and antimicrobial activity of the essential oil of Anthemis altissima L. var. altissima. Natural Product Research 26(20):1931-1934.

Santos PL, Matos JPSCF, Picot L, Almeida JRGS, Quintans JSS, Quintans-Junior LL. (2019). Citronellol, a monoterpene alcohol with promising pharmacological activities - A systematic review. Food and Chemical Toxicology 123:459-469.

Song MK, Um JY, Jang HJ, Lee BC (2012). Beneficial effect of dietary Ephedra sinica on obesity and glucose intolerance in high-fat diet-fed mice. Experimental and Therapeutic Medicine 3(4):707-712.

Stewart HH (1929). The use of ephedrine in asthma and whooping-cough. British Medical Journal 1(3554):293.

Sukatar A, Karabay-Yavaşsoglu N, Ozdemir G, Horzum Z (2006). Antimicrobial activity of volatile component and various extracts of Enteromorpha linza (Linnaeus) J. Agardh from the coast of Izmir, Turkey. Annals of Microbiology 56:275-279.

Tang S, Ren J, Kong L, Yan G, Liu C, Han Y, Sun H, Wang XJ (2023). Ephedrae herba: A review of its phytochemistry, pharmacology, clinical application, and alkaloid toxicity. Molecules 28(2):663.

Wang X, Huang M, Peng Y, Yang W, Shi J (2022). Antifungal activity of 1-octen-3-ol against Monilinia fructicola and its ability in enhancing disease resistance of peach fruit. Food Control 108804.

Wong-Paz JE, Contreras-Esquivel JC, Rodríguez-Herrera R, Carrillo-Inungaray ML, López LI, Nevárez-Moorillón GV, Aguilar CN (2015). Total phenolic content, in vitro antioxidant activity and chemical composition of plant extracts from semiarid Mexican region. Asian Pacific Journal of Tropical Medicine 8(2):104-111.

Wooltorton E, Sibbald B (2002). Ephedra/ephedrine: Cardiovascular and CNS effects. Canadian Medical Association Journal 166(5): 633

Yen GC, Chen HY (1995). Antioxidant activity of various tea extracts in relation to their anti-mutagenicity. Journal of Agriculture and Food Chemistry 43:27-37.




How to Cite

YOUNES, K. M., ALHEIBSHY, F., ALRASHIEDI, A. M., ALWHADANI, L. A., ALSHAMMARI, S. M., ALSHAMMARI, A. S., ALBALAWI, M. K., ALMUTIRI, T. A., & ALJOHANI, G. (2024). Insight into possible therapeutic applications of Ephedra transitoria: in-vitro biological, toxicological activities and GC-MS analysis of aerial parts’ extract . Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 52(1), 13442.



Research Articles
DOI: 10.15835/nbha52113442