Inhibition Effect on Antibiotic Resistant Bacteria and Protosappanin B Content in A-Po-That Formula Extract and Stability Testing
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Abstract
Introduction and Objectives: The 21stA-Po-That traditional drug formula exhibits antibacterial activity against many bacteria that cause diarrhea and its main chemical constituents are brazilin and gallic acid. But there have been no studies on its effects on antibiotic-resistant bacteria and its stability in inhibiting antibiotic-resistant bacteria, as well as its other chemical compounds. The objectives of the study were to examine the antibacterial effects of 21stA-Po-That extract against extended-spectrum beta-lactamase-producing Escherichia coli or ESBL E. coli, analyze its chemical composition and assess the stability of the extract under accelerated conditions.
Methods: Extraction of the A-Po-That remedy was performed using a water decoction method. Subsequently, the antibacterial activity of the extract was assessed by the broth microdilution assay, while the chemical composition was quantified using the HPLC approach. The A-Po-That remedy extract was examined for stability under accelerated conditions for 6 months and tested/analyzed for antibacterial activity and chemical compounds.
Results: The 21stA-Po-That remedy extract was found to exhibit antibacterial activity against ESBL E. coli with minimum inhibitory concentration (MIC) values ranging from 0.625 to 2.5 mg/mL. Protosappanin B was the primary chemical constituent of the A-Po-That extract with a chemical concentration of 2.89±0.21%w/w. The stability testing revealed that the A-Po-That extract exhibited stability in terms of its antibacterial activity and a change in its protosappanin B content after 120 days of testing. However, there was no significant difference in the content of protosappanin B between day 180 and day 0.
Discussion: The 21stA-Po-That remedy extract effectively suppressed the growth of ESBL E. coli. However, the primary chemical compound identified was protosappanin B, which differs in contrast to what was previously reported. Seasonal and climatic changes may have an impact on the chemical composition of the remedy’s medicinal plant ingredients. Furthermore, the A-Po-That extract exhibited consistent antibacterial activity, but displayed different results in terms of chemical composition. The changes in moisture and temperature might be contributing to the changing chemical composition of the remedy.
Conclusions and recommendation: The findings indicate that the A-Po-That formula extract has the ability to inhibit ESBL E. coli growth with protosappanin B being identified as its main chemical compound. After undergoing stability testing for 180 days, the concentration of protosappanin B remained unchanged compared to day 0. However, the concentration of protosappanin B exhibited increased after 120 days. Thus, it is necessary to store A-Po-That extract in conditions of low temperature and low humidity to prevent any alteration of its chemical elements. Before performing any efficacy study of the A-Po-That formula in patients, it is necessary to perform a toxicity study first.
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References
World Health Organization (WHO). Antimicrobial resistance [Internet]. 2023. [cited 2024 May 3]. Available from: https://www.who.int/news-room/fact-sheets/detail/antimicrobial-resistance.
Rossolini G, Arena F, Pecile P, Pollini S. Update on the antibiotic resistance crisis. Current Opinion in Pharmacology. 2014;18:56-60.
Hawser SP, Bouchillon SK, Hoban DJ, Badal RE. In vitro susceptibilities of aerobic and facultative anaerobic gram-negative bacilli from patients with intra-abdominal infections worldwide from 2005–2007: results from the SMART study. International journal of antimicrobial agents. 2009;34(6):585-8.
Sawatwong P, Sapchookul P, Whistler T, Gregory CJ, Sangwichian O, Makprasert S, Jorakate P, Srisaengchai P, Thamthitiwat S, Promkong C, Nanvatthanachod P, Vanaporn M, Rhodes J. High burden of extended-spectrum β-lactamase–producing Escherichia coli and Klebsiella pneumoniae bacteremia in older adults: A seven-year study in two rural Thai provinces. American Journal of Tropical Medicine and Hygiene. 2019;100(4):943-51.
Wongut-sa P. Extended-spectrum β-lactamases producing in Escherichia coli and Klebsiella pneumoniae isolated from specimens of patients in Ratchaburi hospital (2009-2013). Region 4 Medical Journal. 2015;17(3):222-9. (in Thai)
Karaket C. The prevalence of extended spectrum beta-lactamase producing in K.pneumoniae and E.coli at Chiang Kham hospital during 2011-2012. Phichit hospital journal. 2012;28:21-30. (in Thai)
Duksukkaew S. The prevalence of extended spectrum beta-lactamase producing in K. pneumoniae and E. coli at Phatthalung hospital. Region 11 Medical Journal. 2016;30(2):97-103. (in Thai)
Chomjan T, Singto P. Multiple-antimicrobial resistance (AMR) of patients in internal medicine department APHEIT. Journal of Nursing and Health. 2022;4(3):1-16. (in Thai)
Murray C, Ikuta KS, Sharara F, Swetschinski L, Robles Aguilar G, Gray A. Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis. The Lancet. 2022;399(10325):629-55.
Ayurveda Thamrong Faculty of Medicine Siriraj Hospital Mahidol university. The textbook of Thai traditional medicine. Bangkok: Supavanit printing; 2007. 309 p. (in Thai)
Department of Thai Traditional and Alternative Medicine, Ministry of Public Health. Book on the medicine of King Narai. Bangkok: The War Veterans Organization of Thailand Printing; 1999. 180 p. (in Thai)
Panthong S, Sakpakdeejaroen I, Kuropakornpong P, Jaicharoensub J, Itharat A. Antibacterial activity and stability evaluation of “Apo-taat” remedy extract for inhibiting diarrhoea-causing bacteria. Tropical Journal of Natural Product Research. 2020;4(12):1101-7.
Bureau of Drug and Narcotic Department of Medical Sciences, Ministry of Public Health. Thai herbal pharmacopoeia 2021. Nonthaburi: Department of Medical Sciences, Ministry of Public Health; 2023
Tang W, Eisenbrand G. Caesalpinia sappan L. In: Chinese Drugs of Plant Origin. Berlin: Springer; 1992.
Zanin JLB, De Carvalho BA, Salles Martineli P, Dos Santos MH, Lago JHG, Sartorelli P, Viegas C, Soares MG. The genus Caesalpinia L.(Caesalpiniaceae): phytochemical and pharmacological characteristics. Molecules. 2012;17(7):7887-902.
Pattananandecha T, Apichai S, Julsrigival J, Ogata F, Kawasaki N, Saenjum C. Antibacterial activity against foodborne pathogens and inhibitory effect on anti-inflammatory mediators' production of brazilin-enriched extract from Caesalpinia sappan Linn. Plants (Basel, Switzerland). 2022;11(13):1698.
Batubara I, Mitsunaga T, Ohashi H. Brazilin from Caesalpinia sappan wood as an antiacne agent. Journal of Wood Science. 2010;56:77–81.
Monroy LV, Cauich JC, Ortega AM, Campos MS. Medicinal plants as potential functional foods or resources for obtaining anticancer activity metabolites. Oncological Functional Nutrition. Cambridge: Academic Press; 2021. p. 161-94.
Rajamanickam K, Yang J, Sakharkar MK. Gallic acid potentiates the antimicrobial activity of tulathromycin against two key bovine respiratory disease (BRD) causing-pathogens. Frontiers in pharmacology. 2019;9:1486.
De R, Sarkar A, Ghosh P, Ganguly M, Karmakar BC, Saha DR, Halder A, Chowdhury A, Mukhopadhyay AK. Antimicrobial activity of ellagic acid against Helicobacter pylori isolates from India and during infections in mice. The Journal of antimicrobial chemotherapy, 2018;73(6):1595–603.
Vu TT, Kim H, Tran VK, Vu HD, Hoang TX, Han JW, Choi YH, Jang KS, Choi GJ, Kim, JC. Antibacterial activity of tannins isolated from Sapium baccatum extract and use for control of tomato bacterial wilt. PloS one. 2017;12(7):e0181499.
Herbal Product Division, Food and Drug Administration. Guidline of apply for herbal products permission [Internet]. 2023. [cited 2024 Jan 25]. Available from: https://herbal.fda.moph.go.th/product/category/law-regis/. (in Thai)
Sarker SD, Nahar L, Kumarasamy Y. Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth, and its application in the in vitro antibacterial screening of phytochemicals. Methods. 2007;42(4):321-4.
Kondo S, Apisarnthanarak A, Trakulsomboon S, Bootkotr W, Mingmalairak C, Mahawongkajit P, Juntong J, Limpavitayaporn P, Sriussadaporn E, Tongyoo A, Boonyasatid P, Chunsirisub T, Nakornchai K, Thowprasert W, Kiratisin P, Palittapongarnpim P. Prevalence of extended-spectrum β-lactamase-producing enterobacterales and distribution of blaESBL genes from patients who underwent abdominal surgery. Science & Technology Asia. 2022;27(4):104–16.
European Society of clinical microbiolagy and infectious diseases. EUCAST clinical breakpoint tables [Internet]. 2024 [cited 2024 Oct 19]; Available from: https://www.eucast.org/clinical_breakpoints.
ICH Harmonised Tripartite guideline. Validation of analytical procedures: text and methodology. Q2 (R1). 2005;1(20):05.
International Conference on Harmonization. Q1A(R2): Stability testing of new drug substances and products [Internet]. 2003. [cited 2024 Feb 02]; Available from: https://www.ich.org/products/guidelines.html.
Srichumporn W, Chaisowwong W, Intanon M, Na-Lampang K. Extended-spectrum beta-lactamase-producing Escherichia coli from pork in Muang district, Chiang Mai Province, Thailand. Veterinary world. 2022;15(12):2903–9.
Wang Z, Sun J-B, Qu W, Guan F-Q, Li L-Z, Liang J-Y. Caesappin A and B, two novel protosappanins from Caesalpinia sappan L. Fitoterapia. 2014;92:280-4.
O’Donnell G, Bucar F, Gibbons S. Phytochemistry and antimycobacterial activity of Chlorophytum inornatum. Phytochemistry. 2006;67(2):178-82.
Górniak I, Bartoszewski R, Króliczewski J. Comprehensive review of antimicrobial activities of plant flavonoids. Phytochemistry Reviews. 2019;18:241-72.
Zuo GY, Han ZQ, Han J, Hao XY, Tang HS, Wang GC. Antimicrobial activity and synergy of antibiotics with two biphenyl compounds, protosappanins A and B from Sappan lignum against methicillin-resistant Staphylococcus aureus strains. Journal of Pharmacy and Pharmacology. 2015;67(10):1439-47.
Aires A, Fernandes C, Carvalho R, Bennett RN, Saavedra MJ, Rosa EA. Seasonal effects on bioactive compounds and antioxidant capacity of six economically important brassica vegetables. Molecules. 2011;16(8):6816-32.
European Committee for Antimicrobial Susceptibility Testing (EUCAST) of the European Society of Clinical Microbiology and Infectious Diseases (ESCMID). Determination of minimum inhibitory concentrations (MICs) of antibacterial agents by agar dilution. EUCAST E Def. 2000;6(9):509-15.
Donkor MN, Donkor AM, Mosobil R. Combination therapy: synergism among three plant extracts against selected pathogens. BMC Research Notes. 2023;16(1):1-5.
Tian Q, Wei S, Su H, Zheng S, Xu S, Liu M, Bo R, Li J. Bactericidal activity of gallic acid against multi-drug resistance Escherichia coli. Microbial Pathogenesis. 2022;173(Pt A):105824.
Nirmal NP, Rajput MS, Prasad RG, Ahmad M. Brazilin from Caesalpinia sappan heartwood and its pharmacological activities: A review. Asian Pacific Journal of Tropical Medicine. 2015;8(6):421-30.
Warinhomhaun S, Sritularak B, Charnvanich D. A simple high-performance liquid chromatographic method for quantitative analysis of brazilin in Caesalpinia sappan L. extracts. Thai Journal of Pharmaceutical Sciences. 2018;42(4):208-13.
Insuan W, Sillawatthumrong N, Chahomchuen T, Khamchun S, Chueahongthong F, Insuan O. Brazilin content and potential biological properties of Caesalpinia sappan L. heartwood extracts from different extraction methods. Discover Applied Sciences. 2024;6:509.
Xia Z, Li D, Li Q. Zhang Y, Kang W. Simultaneous determination of brazilin and protosappanin B in Caesalpinia sappan by ionic-liquid dispersive liquid-phase microextraction method combined with HPLC. Chemistry Central Journal. 2017;11:114.
Chaphalkar R, Apte KG, Talekar Y, Ojha SK, Nandave M. Antioxidants of Phyllanthus emblica L. Bark extract provide hepatoprotection against ethanol-induced hepatic damage: a comparison with silymarin. Oxidative medicine and cellular longevity. 2017;3876040.