Isolation and Identification of Phenolic Compounds in Guava Leaves and Assessment of their Cytotoxic Effects Against AMJ-13 and MCF-7 Breast Cancer Cell Lines
DOI:
https://doi.org/10.32007/jfacmedbaghdad.6632326Keywords:
Caffeic acid, Gallic acid, Guava, Luteolin, Ultrasonic-assisted extractionAbstract
Background: Psidium guajava, commonly known as guava, is a tropical tree prized for its nutritious fruit and medicinal properties. This member of the Myrtacease family is rich in phytochemicals, which are natural compounds with potential health benefits. Studies have shown that guava leaves and fruits possess various pharmacological activities, including anti-cancer properties against several cancer cell lines.
Objectives: This study aimed to isolate and identify phenolic compounds in guava leaves and assess their cytotoxic activity against AMJ-13 and MCF-7 cell lines by ethyl acetate fraction of guava leaves grown in Iraq.
Methods: The researchers will employ High-performance liquid chromatography
(HPLC) and Fourier transform infrared (FTIR) techniques to analyze the guava leaf extract. Subsequently, preparative High-performance liquid chromatography was used to isolate the specific phenolic compounds of interest, and a colorimetric MTT reduction assay, was conducted using guava extract to assess their effects on human cancer cell lines.
Results: The researchers successfully isolated pure samples of caffeic acid, luteolin, and Gallic acid, which are all flavonoids, from the guava leaf extract using preparative High-performance liquid chromatography. These isolated phenolic compounds were from ethyl acetate fraction (F3). This enriched fraction was tested for its cytotoxic activity against the Iraqi AMJ-13 and human MCF7 breast cancer cell lines. The results showed a decrease in cell viability, indicating the fraction’s potential anti-cancer properties. The fraction was more effective against the Iraqi AMJ-13 cells with an IC50 value of 414.3 μg /ml, compared to the human MCF7 cells with an IC50 value of 698.3 μg /ml.
Conclusion: The analytical techniques used in this study, like HPLC and FTIR, provided a detailed profile of the phenolic compounds present in guava leaves. This information, combined with the cytotoxic tests, suggests that guava leaves have the potential to kill cancer cells in a concentration-dependent manner.
Received: Feb.,2024
Revised: August ,2024
Accepted: Sept.,2024
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References
Abd AM, Kadhim EJ. Phytochemical investigation of aerial parts of Iraqi Cardaria draba. Iraqi Journal of Pharmaceutical Sciences (P-ISSN 1683-3597 E-ISSN 2521-3512). 2020 Dec 27;29(2):27-36. https://doi.org/10.31351/vol29iss2pp27-36
Satar_Al_Baaj A, Abdul-Jalil TZ. Phytochemical Screening of Petroleum Ether Fractions by GC/MS and Isolation of Lupeol from Two Different Parts of Iraqi Leucaena leucocephala.(Conference Paper). Iraqi Journal of Pharmaceutical Sciences (P-ISSN 1683-3597 E-ISSN 2521-3512). 2022;31:62-74. https://doi.org/10.31351/vol31isssuppl.pp62-74
Ibrahem NM, Khadum EJ, Mutlag SH. Isolation of Catchin and Epigallocatchin from Iraqi Rhus coriaria By Preparative High-Performance Liquid Chromatography (PHPLC). Iraqi Journal of Pharmaceutical Sciences (P-ISSN 1683-3597 E-ISSN 2521-3512). 2022 Dec 25;31(2):271-82. https://doi.org/10.31351/vol31iss2pp271-282
Kadhim EJ .Determination and Isolation of Valuable Bioactive Compound (lupeol) from Portulacaria afra Jacq. International Journal of Drug Delivery Technology.2023; 13(1);199–204 .https://doi.org/10.25258/ijddt.13.1.30
Khamees AH, Kadhim EJ, Sahib HB, Mutlag SH. In vitro analysis of antioxidant and antimicrobial activity of Iraqi Bryonia dioica. Int J Pharm SciRevRes.2017;43(1):248-52. https://doi.org/10.47583/ijpsrr.2024.v84i01.015
Salman ZO, Alwash BM, Kadhim EJ. Effect of essential oil of Cestrum nocturnum flowers cultivated in Iraq as antioxidant and elongation cold storage period of minced meat. Iraqi Journal of Agricultural Sciences. 2019 Mar 1;50(2). https://doi.org/10.36103/ijas.v52i6.1496
Arévalo-Marín E, Casas A, Landrum L, Shock MP, Alvarado-Sizzo H, Ruiz-Sanchez E, Clement CR. The taming of Psidium guajava: Natural and cultural history of a neotropical fruit. Frontiers in plant science. 2021 Sep 28;12:714763. https://doi.org/10.3389/fpls.2021.714763
Landrum LR. The genus Psidium (Myrtaceae) in the state of Bahia, Brazil. Herbarium, Natural History Collections, School of Life Sciences, Arizona State University; 2017 Aug. https://doi.org/10.11646/phytotaxa.288.2.6
Rouseff RL, Onagbola EO, Smoot JM, Stelinski LL. Sulfur volatiles in Guava (Psidium guajava L .) Leaves : Possible defense mechanism. J Agric FoodChem. 2008;56:8905–10. https://doi.org/10.1021/jf801735v
Gutiérrez RM, Mitchell S, Solis RV. Psidium guajava: A review of its traditional uses, phytochemistry and pharmacology. Journal of ethnopharmacology. 2008 Apr 17;117(1):1-27. https://doi.org/10.1016/j.jep.2008.01.025
Gutiérrez RM, Mitchell S, Solis RV. Psidium guajava: A review of its traditional uses, phytochemistry and pharmacology. Journal of ethnopharmacology. 2008 Apr 17;117(1):1-27. https://doi.org/10.1016/j.jep.2008.01.025
Alonso-Castro A, Jose Maldonado-Miranda J, Zarate-Martinez A, Jacobo-Salcedo M, del R, Fernández-Galicia C, et al. Medicinal plants used in the huasteca potosina, México. J Ethnopharmacol. (2012) 143:292–8. https://doi.org/10.1016/j.jep.2012.06.035
Juárez-Vázquez MC, Carranza-Álvarez C, Alonso-Castro AJ, González-Alcaraz VF, Bravo-Acevedo E, Chamarro-Tinajero FJ, et al. Ethnobotany of medicinal plants used in xalpatlahuac, guerrero, México. J Ethnopharmacol. (2013) 148:521–7. https://doi.org/10.1016/j.jep.2013.04.048
Omayio DG, Abong GO, Okoth MW, Gachuiri CK, Mwang'ombe AW. Current status of guava (Psidium guajava L.) production, utilization, processing and preservation in Kenya: a review. Cur Agric Res J. (2019) 7:318–31. https://doi.org/10.12944/CARJ.7.3.07
Gutiérrez RMP, Mitchell S, Solis RS. Psidium guajava: Areview of its traditional uses, photochemistry and pharmacology. Journal of Ethnopharmacology. 2008;117(1):127. https://doi.org/10.1016/j.jep.2008.01.025
Kamath JV, Rahul N, Ashok Kumar CK. MohanaLakshmi S. P. guajava L: A review, International Journal of Green Pharmacy. 2008; 2(1):9-12. https://doi.org/10.4103/0973-8258.39155
Lok B, Babu D, Tabana Y, Dahham SS, Adam MA, Barakat K, Sandai D. The Anticancer Potential of Psidium guajava (Guava) Extracts. Life. 2023 Jan 28;13(2):346. https://doi.org/10.3390/life13020346
Bazioli JM, Costa JH, Shiozawa L, Ruiz AL, Foglio MA, Carvalho JE. Anti-estrogenic activity of guajadial fraction, from guava leaves (Psidium guajava L.). Molecules. 2020 Mar 27;25(7):1525. https://doi.org/10.3390/molecules25071525
Gao Y, Li GT, Li Y, Hai P, Wang F, Liu JK. Guajadials CF, four unusual meroterpenoids from Psidium guajava. Natural Products and Bioprospecting. 2013 Feb;3:14-9. https://doi.org/10.1007/s13659-012-0102-4
Abdlkareem SK. Isolation, Identification, and Quantification of Two Compounds from Cassia glauca Cultivated in Iraq. Iraqi Journal of Pharmaceutical Sciences (P-ISSN 1683-3597 E-ISSN 2521-3512). 2023 Dec 30;32(3):95-104. https://doi.org/10.31351/vol32iss3pp95-104
Ghalib SA, Kadhim EJ. The Investigation of Some Phytochemical Compounds Found in Anchusa strigosa L. Grown Naturally in Iraq. Iraqi Journal of Pharmaceutical Sciences (P-ISSN 1683-3597 E-ISSN 2521-3512). 2021 Jun 19;30(1):179-88. https://doi.org/10.31351/vol30iss1pp179-188
Tawfeeq AA, Ali SH. Isolation and Structural Characterization of Quercetin 3-O-Rhamnoside and Essential oil Estimation from Leaves of Iraqi Cupressus sempervirens L (Conference Paper). Iraqi Journal of Pharmaceutical Sciences 2022;31(:121-30. https://doi.org/10.31351/vol31isssuppl.pp121-130
Jewely HM, Zuhair T. Evaluation of Antileishmanial Activity of Osteospermum ecklonis Extract of Aerial Parts against Leishmania donovani: in vitro (Conference Paper). Iraqi Journal of Pharmaceutical Sciences (P-ISSN 1683-3597 E-ISSN 2521-3512). 2022;31(Suppl.):45-53. https://doi.org/10.31351/vol31isssuppl.pp45-53
Khamees AH. Isolation, characterization and quantification of a pentacyclic triterpinoid compound ursolic acid in Scabiosa palaestina L. distributed in the north of Iraq. Plant Science Today. 2022 Jan 1;9(1):178-82. https://doi.org/10.14719/pst.1398
Ibrahem NM, Khadum EJ, Mutlag SH. Isolation of Catchin and Epigallocatchin From Iraqi Rhus coriaria By Preparative High-Performance Liquid Chromatography (PHPLC). Iraqi Journal of Pharmaceutical Sciences (P-ISSN 1683-3597 E-ISSN 2521-3512). 2022 Dec 25;31(2):271-82. https://doi.org/10.31351/vol31iss2pp271-282
Shah SM, Abdul-Jalil TZ. Qualitative and Quantitative Estimation or Chemical Constituents from Leaves and Roots of Iraqi Agave Attenuata by GC-MS and RP-HPLC (Conference Paper). Iraqi Journal of Pharmaceutical Sciences (P-ISSN 1683-3597 E-ISSN 2521-3512). 2022;31(Suppl.):75-85. https://doi.org/10.31351/vol31isssuppl.pp75-85
Freshney RI. Culture of animal cells: a manual of basic technique and specialized applications. John Wiley & Sons; 2015 Dec 23. https://doi.org/10.1002/9780470649367
Ryu NH, Park KR, Kim SM, Yun HM, Nam D, Lee SG, Jang HJ, Ahn KS, Kim SH, Shim BS, Choi SH. A hexane fraction of guava leaves (Psidium guajava L.) induces anticancer activity by suppressing AKT/mammalian target of rapamycin/ribosomal p70 S6 kinase in human prostate cancer cells. Journal of medicinal food. 2012 Mar 1;15(3):231-41. https://doi.org/10.1089/jmf.2011.1701
Ashraf A, Sarfraz RA, Rashid MA, Mahmood A, Shahid M, Noor N. Chemical composition, antioxidant, antitumor, anticancer and cytotoxic effects of Psidium guajava leaf extracts. PharmaceuticalBiology.2016;2;54(10):1971-81.https://doi.org/10.3109/13880209.2015.1137604
Buqui GA, Gouvea DR, Sy SK, Voelkner A, Singh RS, da Silva DB, Kimura E, Derendorf H, Lopes NP, Diniz A. Pharmacokinetic evaluation of avicularin using a model-based development approach. Planta Medica. 2015;81(05):373-81. https://doi.org/10.1055/s-0035-1545728
Gahlaut A, Chhillar AK. Evaluation of antibacterial potential of plant extracts using resazurin based microtiter dilution assay. International Journal of Pharmacy and Pharmaceutical Sciences. 2013 Apr;5(2):372-6. https://doi.org/10.1186/s12866-014-0259-6
Soule HD, Vazguez J, Long A, Albert S, Brennan M. A human cell line from a pleural effusion derived from a breast carcinoma. J Natl Cancer Inst. 1973 Nov;51(5):1409-16. doi: 10.1093/jnci/51.5.1409. PMID: 43577 https://doi.org/10.1111/j.1749-0774.2008.00056.x.
Ashraf A, Sarfraz RA, Rashid MA, Mahmood A, Shahid M, Noor N. Chemical composition, antioxidant, antitumor, anticancer and cytotoxic effects of Psidium guajava leaf extracts. PharmaceuticalBiology.2016;2;54(10):1971-81. https://doi.org/10.3109/13880209.2015.1137604
Buqui GA, Gouvea DR, Sy SK, Voelkner A, Singh RS, da Silva DB, Kimura E, Derendorf H, Lopes NP, Diniz A. Pharmacokinetic evaluation of avicularin using a model-based development approach. Planta Medica. 2015;81(05):373-81. https://doi.org/10.1055/s-0035-1545728
Gahlaut A, Chhillar AK. Evaluation of antibacterial potential of plant extracts using resazurin based microtiter dilution assay. International Journal of Pharmacy and Pharmaceutical Sciences. 2013 Apr;5(2):372-6. https://doi.org/10.1186/s12866-014-0259-6
Gao T, Ci Y, Jian H, An C. FTIR investigation of the interaction of tumor cells treated with caffeic acid and chlorogenic acid. Vibrational spectroscopy. 2000 Dec 1;24(2):225-31. https://doi.org/10.1016/s0924-2031(00)00091-6
Rajhard S, Hladnik L, Vicente FA, Srčič S, Grilc M, Likozar B. Solubility of luteolin and other polyphenolic compounds in water, nonpolar, polar aprotic and protic solvents by applying ftir/hplc. Processes. 2021 Oct 30;9(11):1952. https://doi.org/10.3390/pr9111952
Werner I, Bacher A, Eisenreich W. Retrobiosynthetic NMR studies with 13C-labeled glucose: formation of gallic acid in plants and fungi. Journal of Biological Chemistry. 1997 Oct 10;272(41):25474-82. https://doi.org/10.1074/jbc.272.41.25474
Tena-Rojas KF, Martínez-Flores HE, Garnica-Romo MG, de Dios Figueroa-Cárdenas J, Meléndez-Herrera E, Salgado-Garciglia R. Influence of factors and interactions in ultrasound-assisted extraction and conventional maceration on aqueous extract of Psidium guajava leaves. Bioactive Compounds in Health and Disease-Online ISSN: 2574-0334; Print ISSN: 2769-2426. 2022 Oct 31;5(10):186-201. https://doi.org/10.31989/bchd.v5i10.969
Kancherla N, Dhakshinamoothi A, Chitra K, Komaram RB. Preliminary analysis of phytoconstituents and evaluation of anthelminthic property of Cayratia auriculata (in vitro). Maedica. 2019 Dec;14(4):350. https://doi.org/10.26574/maedica.2019.14.4.350
Sun L, Jin HY, Tian RT, Wang MJ, Liu LN, Ye LP, Zuo TT, Ma SC. A simple method for HPLC retention time prediction: linear calibration using two reference substances. Chinese Medicine. 2017 Dec;12:1-2. https://doi.org/10.1186/s13020-017-0137-x
Ghasemi M, Turnbull T, Sebastian S, Kempson I. The MTT assay: utility, limitations, pitfalls, and interpretation in bulk and single-cell analysis. International journal of molecular sciences. 2021 Nov 26;22(23):12827. https://doi.org/10.3390/ijms222312827
Tan MC, Carranza MS, Linis VC, Malabed RS, Oyong GG. Antioxidant, cytotoxicity, and antiophidian potential of Alstonia macrophylla bark. ACS omega. 2019 May 30;4(5):9488-96. https://doi.org/10.1021/acsomega.9b00082
Abdel-Aal MT, Zahran F, Banawy MA. Antitumor effect of guava leaves extracts on Ehrlich Ascites carcinoma in mice in vitro and in vivo. Biochemistry Letters. 2022 Dec 1;18(1):119-39. https://doi.org/10.21608/blj.2022.285143
Kemegne GA, Bettache N, Nyegue MA, Etoa FX, Menut C. Cytotoxic activities of Psidium guajava and Mangifera indica plant extracts on human healthy skin fibroblasts and human hepatocellular carcinoma. Issues in Biological Sciences and Pharmaceutical Research. 2020;8(4):58-64. https://doi.org/10.1177/1934578x1801300730
Prakoso NI, Nita MT. Exploring anticancer activity of the Indonesian guava leaf (Psidium guajava L.) fraction on various human cancer cell lines in an in vitro cell-based approach. Open Chemistry. 2023 Aug 4;21(1):20230101. https://doi.org/10.1515/chem-2023-0101
Soule HD, Vazguez J, Long A, Albert S, Brennan M. A human cell line from a pleural effusion derived from a breast carcinoma. J Natl Cancer Inst. 1973 Nov;51(5):1409-16. doi: 10.1093/jnci/51.5.1409. PMID: 43577 https://doi.org/10.1111/j.1749-0774.2008.00056.x
Sahin F., Kannangai R., Adegbola O., Wang J., Su G., Torbenson M. mTOR and P70 S6 kinase expression in primary liver neoplasms. Clin. Cancer Res. 2004;10:8421–8425. https://doi.org/10.1158/1078-0432.CCR-04-0941
Xu G., Zhang W., Bertram P., Zheng X.F., McLeod H. Pharmacogenomic profiling of the PI3K/PTEN-AKTmTOR pathway in common human tumors. Int. J. Oncol. 2004;24:893–900. https://doi.org/10.3892/ijo.24.4.893
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