Phytochemical profile, antioxidant potential, and antibacterial activity of Thai polyherbal formulation, a Ya Kae Rok Bit remedy

Main Article Content

CHAWALIT YONGRAM
PALAPON CHIMPALEE
SUWADEE CHOKCHAISIRI
SASIPEN KRUTCHANGTHONG
JIRAPORN SONGSRI
THAVATCHAI KAMOLTHAM
PANUPAN SRIPAN

Abstract

Abstract. Yongram C, Chimpalee P, Chokchaisiri S, Krutchangthong S, Songsri J, Kamoltham T, Sripan P. 2026. Phytochemical profile, antioxidant potential, and antibacterial activity of Thai polyherbal formulation, a Ya Kae Rok Bit remedy. Biodiversitas 27 (5): d270516. https://doi.org/10.13057/biodiv/d270516. This study investigated the quantitative and qualitative phytochemical diversity and biological activities of Ya Kae Rok Bit (YKRB) extracts prepared using different solvents (hexane, ethyl acetate, and ethanol). Total phenolic, flavonoid, and tannin contents were also evaluated. Antioxidant capacity was evaluated using DPPH, ABTS, and FRAP assays, while antibacterial activity against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923 was determined by disc diffusion and broth microdilution. GC-MS analysis identified 28 phytochemical constituents, including neutral cannabinoids, with tetrahydrocannabinol (23.24-27.91%) and cannabinol (14.18-21.06%) as relatively abundant compounds based on peak area normalization, together with macelignan (6.34-10.55%). HPLC confirmed the highest Δ9-THC content in the hexane extract (6.92 mg/g extract). The extracts showed measurable antioxidant activity in the DPPH assay (IC50 = 56.72-95.25 µg/mL) and stronger radical scavenging activity in the ABTS assay (IC50 = 23.26-26.93 µg/mL), with reducing capacity observed in the FRAP assay (200.92 mmol Fe²⁺/100 g extract). Moreover, YKRB showed antibacterial activity against S. aureus (MIC = 25 mg/mL), while no activity was observed against E. coli. These findings demonstrate solvent-dependent phytochemical variation and provide preliminary evidence of antioxidant and selective antibacterial activities of YKRB extracts.

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Alves FS, Cruz JN, de Farias Ramos IN, do Nascimento Brandão DL, Queiroz RN, da Silva GV, da Silva GV, Dolabela MF, da Costa ML, Khayat AS, de Arimatéia Rodrigues do Rego J. 2023. Evaluation of antimicrobial activity and cytotoxicity effects of extracts of Piper nigrum L. and piperine. Separations 10 (1): 21. https://doi.org/10.3390/separations10010021.

Andre CM, Hausman JF, Guerriero G. 2016. Cannabis sativa: The plant of the thousand and one molecules. Front Plant Sci 7: 19. https://doi.org/10.3389/fpls.2016.00019.

Appendino G, Gibbons S, Giana A, Pagani A, Grassi G, Stavri M, Smith E, Rahman MM. 2008. Antibacterial cannabinoids from Cannabis sativa: A structure-activity study. J Nat Prod 71 (8): 1427-1430. https://doi.org/10.1021/np8002673.

Asyhar R, Minarni M, Arista RA, Nurcholis W. 2023. Total phenolic and flavonoid contents and their antioxidant capacity of Curcuma xanthorrhiza accessions from Jambi. Biodiversitas 24 (9): 5007-5014. https://doi.org/10.13057/biodiv/d240944.

Atalay S, Jarocka-Karpowicz I, Skrzydlewska E. 2019. Antioxidative and anti-inflammatory properties of cannabidiol. Antioxidants 9 (1): 21. https://doi.org/10.3390/antiox9010021.

Balouiri M, Sadiki M, Ibnsouda SK. 2016. Methods for in vitro evaluating antimicrobial activity: A review. J Pharm Anal 6 (2): 71-79. https://doi.org/10.1016/j.jpha.2015.11.005.

Bhatwalkar SB, Mondal R, Krishna SBN, Adam JK, Govender P, Anupam R. 2021. Antibacterial properties of organosulfur compounds of garlic (Allium sativum). Front Microbiol 12: 613077. https://doi.org/10.3389/fmicb.2021.613077.

Blaskovich MAT, Kavanagh AM, Elliott AG, Zhang B, Ramu S, Amado M, Lowe GJ, Hinton AO, Pham DMT, Zuegg J, Beare N, Quach D, Sharp MD, Pugliano J, Rogers AP, Lyras D, Tan L, West NP, Crawford DW, Peterson ML, Callahan M, Thurn M. 2021. The antimicrobial potential of cannabidiol. Commun Biol 4: 7. https://doi.org/10.1038/s42003-020-01530-y.

Burgberger M, Mierziak J, Augustyniak B, Wojtasik W, Kulma A. 2025. The power of lignans: Plant compounds with multifaceted health-promoting effects. Metabolites 15 (9): 589. https://doi.org/10.3390/metabo15090589.

Burt S. 2004. Essential oils: Their antibacterial properties and potential applications in foods. Int J Food Microbiol 94 (3): 223-253. https://doi.org/10.1016/j.ijfoodmicro.2004.03.022.

Bush K, Bradford PA. 2016. β-Lactams and β-lactamase inhibitors: An overview. Cold Spring Harb Perspect Med 6 (8): a025247. https://doi.org/10.1101/cshperspect.a025247

Camilleri M. 2018. Cannabinoids and gastrointestinal motility: Pharmacology, clinical effects, and potential therapeutics in humans. Neurogastroenterol Motil 30 (9): e13370. https://doi.org/10.1111/nmo.13370.

Chokchaisiri S, Ngivprom U, Phatthanaphong P, Boon-orn K, Wongsonthom S, Chimpalee P. 2026. Phytochemical profiles and antioxidant activities of four Cannabis sativa cultivars in Thailand. Biodiversitas 27 (1): 1-8. https://doi.org/10.13057/biodiv/d270100.

Christodoulou MC, Orellana Palacios JC, Hesami G, Jafarzadeh S, Lorenzo JM, Domínguez R, Moreno A, Hadidi M. 2022. Spectrophotometric methods for measurement of antioxidant activity in food and pharmaceuticals. Antioxidants (Basel) 11 (11): 2213. https://doi.org/10.3390/antiox11112213.

Cosme F, Aires A, Pinto T, Oliveira I, Vilela A, Gonçalves B. 2025. A comprehensive review of bioactive tannins in foods and beverages. Molecules 30 (4): 800. https://doi.org/10.3390/molecules30040800.

Crowley K, Kiraga Ł, Miszczuk E, Skiba S, Banach J, Latek U, Mendel M, Chłopecka M. 2024. Effects of cannabinoids on intestinal motility, barrier permeability and therapeutic potential in gastrointestinal diseases. Intl J Mol Sci 25 (12): 6682. https://doi.org/10.3390/ijms25126682.

Department of Thai Traditional and Alternative Medicine. 2021. Collection of Conserved Thai Traditional Medicine Wisdom: National Thai Medicine Formulas Incorporating Cannabis. Ministry of Public Health, Nonthaburi.

Do QD, Angkawijaya AE, Tran-Nguyen PL, Huynh LH, Soetaredjo FE, Ismadji S, Ju YH. 2014. Effect of extraction solvent on total phenol content and antioxidant activity. J Food Drug Anal 22 (3): 296-302. https://doi.org/10.1016/j.jfda.2013.11.001.

Farha MA, El-Halfawy OM, Gale RT, MacNair CR, Carfrae LA, Zhang X, Jentsch NG, Magolan J, Brown ED. 2020. Uncovering the hidden antibiotic potential of Cannabis. ACS Infect Dis 6 (3): 338-346. https://doi.org/10.1021/acsinfecdis.9b00419.

Gomes CL, Silva CCAR, Melo CG, Ferreira MRA, Soares LAL, DA Silva RMF, Rolim LA, Rolim Neto PJ. 2021. Development of an analytical method for determination of polyphenols and total tannins from leaves of Syzygium cumini L. Skeels. An Acad Bras Cienc 93 (2): e20190373. https://doi.org/10.1590/0001-3765202120190373.

Jin S, Xu H, Yang C, O K. 2024. Regulation of oxidative stress in the intestine of piglets after enterotoxigenic Escherichia coli (ETEC) infection. Biochim Biophys Acta Mol Cell Res 1871 (5): 119711. https://doi.org/10.1016/j.bbamcr.2024.119711.

Jurowski K, Kobylarz D, Noga M. 2025. ADME profile of BZP (benzylpiperazine) - first application of multi-in silico approach methodology for comprehensive prediction of ADME profile (absorption, distribution, metabolism and excretion) important for clinical toxicology and forensic purposes. Chem Biol Interact 421: 111775. https://doi.org/10.1016/j.cbi.2025.111775.

Khoddami A, Wilkes MA, Roberts TH. 2013. Techniques for analysis of plant phenolic compounds. Molecules 18 (2): 2328-2375. https://doi.org/10.3390/molecules18022328.

Liang J, Dai W, Liu C, Wen Y, Chen C, Xu Y, Huang S, Hou S, Li C, Chen Y, Wang W, Tang H. 2024. Gingerenone A attenuates ulcerative colitis via targeting IL-17RA to inhibit inflammation and restore intestinal barrier function. Adv Sci (Weinh) 11 (28): e2400206. https://doi.org/10.1002/advs.202400206.

Maher C, Hassan KA. 2023. The gram-negative permeability barrier: Tipping the balance of the in and the out. mBio 14 (6): e0120523. https://doi.org/10.1128/mbio.01205-23.

Matulyte I, Jekabsone A, Jankauskaite L, Zavistanaviciute P, Sakiene V, Bartkiene E, Ruzauskas M, Kopustinskiene DM, Santini A, Bernatoniene J. 2020. The essential oil and hydrolats from Myristica fragrans seeds with magnesium aluminometasilicate as excipient: Antioxidant, antibacterial, and anti-inflammatory activity. Foods 9 (1): 37. https://doi.org/10.3390/foods9010037.

Ministry of Public Health. 2016. The Announcement to Define the Thailand National Traditional Medicine Textbook and Thailand National Traditional Pharmacopoeia. Department for Development of Thai Traditional and Alternative Medicine, Ministry of Public Health, Bangkok.

Mokoena D, George BP, Abrahamse H. 2022. The role of Cannabis species on oxidative stress in cancer cells. In: Chakraborti S (eds). Handbook of Oxidative Stress in Cancer. Springer, Singapore. https://doi.org/10.1007/978-981-16-5422-0_201.

Oktiansyah R, Elfita, Widjajanti H, Salni, Setiawan A. 2023. Antibacterial and antioxidant activity of endophytic fungi isolated the petiole of sungkai plant (Peronema canescens). Biodiversitas 24 (12): 6516-6526. https://doi.org/10.13057/biodiv/d241213.

Pellati F, Brighenti V, Sperlea J, Marchetti L, Bertelli D, Benvenuti S. 2018. New methods for comprehensive analysis of bioactive compounds in Cannabis sativa. Molecules 23 (10): 2639. https://doi.org/10.3390/molecules23102639.

Pereira SR, Hackett B, O'Driscoll DN, Sun MC, Downer EJ. 2021. Cannabidiol modulation of oxidative stress and signalling. Neuronal Signal 5 (3): NS20200080. https://doi.org/10.1042/NS20200080.

POWO. 2024. "Plants of the World Online. Facilitated by the Royal Botanic Gardens, Kew. https://powo.science.kew.org/

Rampogu S, Baek A, Gajula RG, Zeb A, Bavi RS, Kumar R, Kim Y, Kwon YJ, Lee KW. 2018. Ginger (Zingiber officinale) phytochemicals gingerenone-A and shogaol inhibit SaHPPK: Molecular docking, molecular dynamics simulations and in vitro approaches. Ann Clin Microbiol Antimicrob 17 (1): 16. https://doi.org/10.1186/s12941-018-0266-9.

Rezvani M. 2024. Oxidative stress-induced gastrointestinal diseases: Biology and nanomedicines-a review. BioChem 4 (3): 189-216. https://doi.org/10.3390/biochem4030010.

Royal Thai Government Gazette. 2019. Narcotics Act No. 7 B.E. 2562 (2019). Government of Thailand, Bangkok.

Rumpf J, Burger R, Schulze M. 2023. Statistical evaluation of DPPH, ABTS, FRAP, and Folin-Ciocalteu assays to assess the antioxidant capacity of lignins. Intl J Biol Macromol 233: 123470. https://doi.org/10.1016/j.ijbiomac.2023.123470.

Saiprom Y, Fakkham S, Promsorn T, Krutchangthong S, Yongram C, Puthongking P. 2026. The effect of solvent extraction on chemical composition, antioxidant, alpha-glucosidase and nitric oxide inhibitor activities of Jindamanee, a Thai traditional formulation. Trop J Nat Prod Res 10 (1): 6778-6784. https://doi:10.26538/tjnpr/v10i1.48.

Stout SM, Cimino NM. 2014. Exogenous cannabinoids as substrates, inhibitors, and inducers of human drug metabolizing enzymes: A systematic review. Drug Metab Rev 46 (1): 86-95. https://doi.org/10.3109/03602532.2013.849268.

Sukweenadhi J, Yunita O, Setiawan F, Kartini, Siagian MT, Danduru AP, Avanti C. 2020. Antioxidant activity screening of seven Indonesian herbal extract. Biodiversitas 21 (5): 2062-2067. https://doi.org/10.13057/biodiv/d210532.

Tang D, Chen M, Huang X, Zhang G, Zeng L, Zhang G, Wu S, Wang Y. 2023. SRplot: A free online platform for data visualization and graphing. PLoS One 18 (11): e0294236. https://doi.org/10.1371/journal.pone.0294236.

Thai Traditional Medicine Institute. 2025. Reference Textbook of Thai Traditional Medicine. Ministry of Public Health, Nonthaburi, Thailand.

World Health Organization (WHO). 2023. Diarrhoeal Disease. WHO, Geneva. https://www.who.int.

Yongram C, Sripan P, Chokchaisiri S, Wonganan O, Meeboonya R, Kamoltham T, Krutchangthong S, Panyatip P. 2025. Antioxidant activity and phytochemical composition of Thai traditional Cannabis longevity recipes: In vitro evaluation of the Phontecho remedy. Trop J Nat Prod Res 9 (12): 6166-6174. https://doi.org/10.26538/tjnpr/v9i12.33.

Zendulka O, Dovrtělová G, Nosková K, Turjap M, Šulcová A, Hanuš L, Juřica J. 2016. Cannabinoids and cytochrome P450 interactions. Curr Drug Metab 17 (3): 206-226. https://doi.org/10.2174/1389200217666151210142051.

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