LCMS/MS analysis and evaluation of anti-inflammatory and antioxidant activities of the polyphenol fraction of Litsea quinqueflora (Dennst.) Suresh

Authors

DOI:

https://doi.org/10.14719/pst.2021.8.4.1243

Keywords:

ABTS, DPPH assay, Free radical scavenging enzymes, HPTLC, LCMS/MS, Protein denaturation

Abstract

The main aim of the work was to scientifically prove the anti-inflammatory property of the polyphenol-rich fraction of hydro-alcoholic leaf extract of Litsea quinqueflora (Dennst.) Suresh by protein denaturation and free radical scavenging activity. The polyphenol-rich fraction of hydro-alcoholic leaf extract was obtained via acid-alkali hydrolysis, followed by fractionation with chloroform and ethyl acetate. HPTLC profiling of the finally obtained ethyl acetate fraction and consequent derivatisation with aluminium chloride revealed the presence of flavonoids in a more purified form. LCMS/MS analysis tentatively identified the presence of bioactive polyphenolic compounds such as gallocatechin, sinapic acid, pinocembrin, paeonol and umbelliferone in the separated fraction. The polyphenol-rich fraction of hydro-alcoholic extract of leaves showed anti-denaturing activity in heat-induced bovine serum albumin denaturation with an IC50 value of 23.59 µg/ml and was statistically significant at 0.1% level. The antioxidant property of the polyphenol-rich fraction determined by its free radical scavenging ability against DPPH and ABTS showed IC50 values 122.98 and 135.44 µg/ml respectively and was also statistically significant at 0.1% level. Hence, the traditional use of Litsea quinqueflora as an anti-inflammatory agent can be attributed to the presence of polyphenols.

Downloads

Download data is not yet available.

References

Abbas M, Saeed F, Anjum FM, Afzaal M, Tufail T, Bashir MS, Ishtiaq A, Hussain S, Suleria HA. Natural polyphenols: An overview. Int J Food Prop. 2017;20(8):1689-99.

Duthie GG, Gardner PT, Kyle JA. Plant polyphenols: are they the new magic bullet?. Proceedings of the Nutrition Society. 2003;62(3):599-03.

Cory H, Passarelli S, Szeto J, Tamez M, Mattei J. The role of polyphenols in human health and food systems: A mini-review. Front Nutr. 2018;5:87.

Yahfoufi N, Alsadi N, Jambi M, Matar C. The immunomodulatory and anti-inflammatory role of polyphenols. Nutr. 2018;10(11):1618-41.

Ambriz-Pérez DL, Leyva-López N, Gutierrez-Grijalva EP, Heredia JB. Phenolic compounds: Natural alternative in inflammation treatment. A Review. Cogent Food Agric. 2016;2(1):1131412. https://doi.org/10.1080/23311932.2015.1131412

Wang YS, Wen ZQ, Li BT, Zhang HB, Yang JH. Ethnobotany, phytochemistry and pharmacology of the genus Litsea: An update. J Ethnopharmacol. 2016;181:66-107. https://doi.org/10.1016/j.jep.2016.01.032

Kong DG, Zhao Y, Li GH, Chen BJ, Wang XN, Zhou HL, Lou HX, Ren DM, Shen T. The genus Litsea in traditional Chinese medicine: an ethnomedical, phytochemical and pharmacological review. J Ethnopharmacol. 2015;164:256-64. https://doi.org/10.1016/j.jep.2015.02.020

Kamle M, Mahato DK, Lee KE, Bajpai VK, Gajurel PR, Gu KS, Kumar P. Ethnopharmacological properties and medicinal uses of Litsea cubeba. Plants. 2019;8(6):150. https://doi.org/10.3390/plants8060150

Liu R, Zhang HC, Zhou F, Wang RM, Tu Q, Wang JY. Flavonoids and alkaloids from the leaves of Litsea fruticosa. Biochem Syst Ecol. 2013;50:293-95. https://doi.org/10.1016/j.bse.2013.04.008

Anilkumar M, Johny J. Evaluation of in vitro anti-Inflammatory activity of the methanolic extract of Litsea quinqueflora (Dennst.) Suresh. J Pharm Biol Sci. 2015;10(2):32-36.

Jose SM, Anilkumar M. In vitro antioxidant activity of Litsea quinqueflora (Dennst.) Suresh. J Pharmacogn Phytochem. 2018;7(4):3217-21.

Rodríguez De Luna SL, Ramírez-Garza RE, Serna Saldívar SO. Environmentally friendly methods for flavonoid extraction from plant material: Impact of their operating conditions on yield and antioxidant properties. Sci World J. 2020. https://doi.org/10.1155/2020/6792069

Czemplik M, Korzun-Ch?opicka U, Szatkowski M, Dzia?o M, Szopa J, Kulma A. Optimization of phenolic compounds extraction from flax shives and their effect on human fibroblasts. Evid-Based Compl Alt Med. 2017. https://doi.org/10.1155/2017/3526392

Yassine EZ, Dalila B, Latifa EM, Smahan B, Lebtar S, Sanae A, Abdellah F. Phytochemical screening, anti-inflammatory activity and acute toxicity of hydro-ethanolic, flavonoid, tannin and mucilage extracts of Lavandula stoechas L. from Morocco. Int J Pharm Phytochem Res. 2016;8(1):31-37.

Lee Y, Howard LR, Villalon B. Flavonoids and antioxidant activity of fresh pepper (Capsicum annuum) cultivars. J Food Sci. 1995;60(3):473-76.

Thangaraj P. Detection of phenolic and flavonoid compounds using high performance thin layer chromatography (HPTLC). In: Pharmacological Assays of Plant-Based Natural Products 2016 (pp. 173-75). Springer, Cham.

Srivasthava MM. (Ed.) HPTLC: High Performance thin- layer chromatography, (2011) Heidelberg, Dordrecht, London, New York: Springer. https://doi.org/10.1007/978-3-642-14025-9_1

Sulaiman CT, Deepak M, Balachandran I. Spectrophotometric and tandem mass spectroscopic analysis of Indian borage (Plectranthus amboinicus (Lour.) Spreng.) for its polyphenolics characterisation. Beni-Suef Univ J Basic Appl Sci. 2018;7(4):471-73.

Mizushima Y, Kobayashi M. Interaction of anti?inflammatory drugs with serum proteins, especially with some biologically active proteins. J Pharm Pharmacol. 1968;20(3):169-73. https://doi.org/10.1111/j.2042-7158.1968.tb09718.x

Kiranmayi GV. Preliminary phytochemical screening and in vitro evaluation of anti-inflammatory, anti-arthritic and thrombolytic activities of ethanolic leaf extract of Bauhinia purpurea. Int J Green Pharm. 2018;12(01). http://dx.doi.org/10.22377/ijgp.v12i01.1627

Brand-Williams W, Cuvelier ME, Berset CL. Use of a free radical method to evaluate antioxidant activity. LWT Food Sci Technol. 1995;28(1):25-30. https://doi.org/10.1016/S0023-6438(95)80008-5

Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorisation assay. Free Radical Biol Med. 1999;26(9-10):1231-37. https://doi.org/10.1016/S0891-5849(98)00315-3

Shunmugaperumal T, Kaur V. In vitro anti-inflammatory and antimicrobial activities of azithromycin after loaded in chitosan-and tween 20-based oil-in-water macroemulsion for acne management. Aaps Pharm Sci Tech. 2016;17(3):700-09. https://doi.org/10.1208/s12249-015-0401-2

Osman NI, Sidik NJ, Awal A, Adam NA, Rezali NI. In vitro xanthine oxidase and albumin denaturation inhibition assay of Barringtonia racemosa L. and total phenolic content analysis for potential anti-inflammatory use in gouty arthritis. Journal of Intercultural Ethnopharmacology. 2016;5(4):343-49. https://doi.org/10.5455/jice.20160731025522

Khoddami A, Wilkes MA, Roberts TH. Techniques for analysis of plant phenolic compounds. Mol. 2013;18(2):2328-75. https://doi.org/10.3390/molecules18022328

Hu J, Zhao Y, Ma C, Wang W, Xing D, Du L. Acid hydrolytic method for determination of Ginkgo biloba total flavonoids in rat plasma by HPLC for pharmacokinetic studies. Tsinghua Sci Technol. 2010;15(4):452-59. https://doi.org/10.1016/S1007-0214(10)70087-0

Alara OR, Abdurahman NH, Ukaegbu CI. Extraction of phenolic compounds: a review. Curr Res Food Sci. 2021;4:200-14. https://doi.org/10.1016/j.crfs.2021.03.011

Wang Q, Ge X, Tian X, Zhang Y, Zhang J, Zhang P. Soy isoflavone: The multipurpose phytochemical. Biomed Rep. 2013;1(5):697-701. https://doi.org/10.3892/br.2013.129

Kaur H, Kaur G. A critical appraisal of solubility enhancement techniques of polyphenols. J Pharm. 2014;1-14. https://doi.org/10.1155/2014/180845

Kuruvilla J, Iyer RS, Anilkumar M. Phytochemical evaluation and HPTLC fingerprint profile of Cissus latifolia Lam. Stem. Int J Pharm Sci Res. 2019;10(2):568-77.

Srivastava B, Sharma VC, Verma SC, Singh R, Jadhav AD. Plant part substitution in Litsea chinensis for medicinal use: A comparative phytochemical approach. Anc Sci Life. 2017; 37(2):57. https://doi.org/10.4103/asl.ASL_168_16

Senguttuvan J, Subramaniam P. HPTLC Fingerprints of Various Secondary Metabolites in the Traditional Medicinal Herb Hypochaeris radicata L. J Bot. 2016;1-11. https://doi.org/10.1155/2016/5429625

Gwatidzo L, Dzomba P, Mangena M. TLC separation and antioxidant activity of flavonoids from Carissa bispinosa, Ficus sycomorus and Grewia bicolar fruits. Nutrire. 2018;43(1):1-7. https://doi.org/10.1186/s41110-018-0062-5

Pontis JA, Costa LA, Silva SJ, Flach A. Color, phenolic and flavonoid content, and antioxidant activity of honey from Roraima, Brazil. Food Sci Technol. 2014;34(1):69-73. https://doi.org/10.1590/S0101-20612014005000015

MassBank; an ecosystem of databases and tools for mass spectrometry reference spectra. Gallocatehin. https://massbank.eu/MassBank/RecordDisplay?id=BS003902

Hossain MB, Rai DK, Brunton NP, Martin-Diana AB, Barry-Ryan C. Characterization of phenolic composition in Lamiaceae spices by LC-ESI-MS/MS. J Agric Food Chem. 2010; 58(19):10576-81. https://doi.org/10.1021/jf102042g

Gamboa-Gómez CI, González-Laredo RF, Gallegos-Infante JA, Pérez M, Moreno-Jiménez MR, Flores-Rueda AG, Rocha-Guzmán NE. Antioxidant and angiotensin-converting enzyme inhibitory activity of Eucalyptus camaldulensis and Litsea glaucescens infusions fermented with kombucha consortium. Food Technol Biotechnol. 2016;54(3):367-74. https://doi.org/10.17113/ftb.54.03.16.4622

Rosales-Castro M, González-Laredo RF, Rocha-Guzmán NE, Gallegos-Infante JA, Rivas-Arreola MJ, Karchesy JJ. Antioxidant activity of fractions from Quercus sideroxyla bark and identification of proanthocyanidins by HPLC-DAD and HPLC-MS. Holzforschung. 2012;66(5):577-84.

MassBank; an ecosystem of databases and tools for mass spectrometry reference spectra. Sinapic acid. https://massbank.eu/MassBank/RecordDisplay?id=PR307086

Sun J, Liang F, Bin Y, Li P, Duan C. Screening non-colored phenolics in red wines using liquid chromatography/ultraviolet and mass spectrometry/mass spectrometry libraries. Mol. 2007;12(3):679-93. https://doi.org/10.3390/12030679

Bystrom LM, Lewis BA, Brown DL, Rodriguez E, Obendorf RL. Characterisation of phenolics by LC–UV/Vis, LC–MS/MS and sugars by GC in Melicoccus bijugatus Jacq. ‘Montgomery’ fruits. Food Chem. 2008;111(4):1017-24. https://doi.org/10.1016/j.foodchem.2008.04.058

Ni?iforovi? N, Abramovi? H. Sinapic acid and its derivatives: natural sources and bioactivity. Compr Rev Food Sci Food Safe. 2014;13(1):34-51. https://doi.org/10.1111/1541-4337.12041

Seal T, Chaudhuri K, Pillai B, Chakrabarti S, Mondal T, Auddy B. Evaluation of antioxidant activities, toxicity studies and the DNA damage protective effect of various solvent extracts of Litsea cubeba fruits. Heliyon. 2020;6(3):e03637. https://doi.org/10.1016/j.heliyon.2020.e03637

MassBank; an ecosystem of databases and tools for mass spectrometry reference spectra. Pinocembrin. https://massbank.eu/MassBank/RecordDisplay.jsp?id=BML00902

Sawaya AC, Tomazela DM, Cunha IB, Bankova VS, Marcucci MC, Custodio AR, Eberlin MN. Electrospray ionisation mass spectrometry fingerprinting of propolis. Anal. 2004;129(8):739-44. https://doi.org/10.1039/B403873H

Simirgiotis MJ, Benites J, Areche C, Sepúlveda B. Antioxidant capacities and analysis of phenolic compounds in three endemic Nolana species by HPLC-PDA-ESI-MS. Mol. 2015; 20(6):11490-507. https://doi.org/10.3390/molecules200611490

Liu R, Zhang HC, Zhou F, Wang RM, Tu Q, Wang JY. Flavonoids and alkaloids from the leaves of Litsea fruticosa. Biochem Syst Ecol. 2013;50:293-95. https://doi.org/10.1016/j.bse.2013.04.008

MassBank; an ecosystem of databases and tools for mass spectrometry reference spectra. Paeonol. https://massbank.eu/MassBank/RecordDisplay?id=PR303539

Zhao X, Sun Y. Analysis of Paeoniae radix by high-performance liquid chromatography-electrospray ionisation-mass spectrometry. Anal Sci. 2003;19(9):1313-15. https://doi.org/10.2116/analsci.19.1313

Kwon MH, Jeong JS, Ryu J, Cho YW, Kang HE. Simultaneous determination of saikosaponin a, paeonol and imperatorin, components of DA-9805, in rat plasma by LC–MS/MS and application to a pharmacokinetic study. J Chromatogr B. 2017; 1068:289-96. https://doi.org/10.1016/j.jchromb.2017.10.059

Gong X, Yang Y, Huang L, Zhang Q, Wan RZ, Zhang P, Zhang B. Antioxidation, anti-inflammation and anti-apoptosis by paeonol in LPS/d-GalN-induced acute liver failure in mice. Int Immunopharmacol. 2017;46:124-32. https://doi.org/10.1016/j.intimp.2017.03.003

MassBank; an ecosystem of databases and tools for mass spectrometry reference spectra. Umbelliferone. https://massbank.eu/MassBank/RecordDisplay?id=FIO00499

Abu-Reidah IM, Ali-Shtayeh MS, Jamous RM, Arráez-Román D, Segura-Carretero A. HPLC–DAD–ESI-MS/MS screening of bioactive components from Rhus coriaria L. (Sumac) fruits. Food Chem. 2015;166:179-91. https://doi.org/10.1016/j.foodchem.2014.06.011

Gajbhiye NA, Makasana J, Dhanani T, Saravanan R. Development and validation of LC—ESI—MS/MS method for simultaneous determination of four coumarin derivatives and an alkaloid from root and stem bark of Aegle marmelos Correa. Acta Chromatographica. 2016;28(4):473-88. https://doi.org/10.1556/1326.2016.28.4.6

Rauf A, Khan R, Khan H, Pervez S, Pirzada AS. In vivo antinociceptive and anti-inflammatory activities of umbelliferone isolated from Potentilla evestita. Nat Prod Res. 2014;28(17):1371-74. https://doi.org/10.1080/14786419.2014.901317

Sangeetha G, Vidhya R. In vitro anti-inflammatory activity of different parts of Pedalium murex (L.). Int J Herb Med. 2016; 4(3):31-36.

Malarkani K, Sarkar I, Selvam S. Denaturation studies on bovine serum albumin–bile salt system: bile salt stabilises bovine serum albumin through hydrophobicity. J Pharmaceut Anal. 2018;8(1):27-36. https://doi.org/10.1016/j.jpha.2017.06.007

Elisha IL, Dzoyem JP, McGaw LJ, Botha FS, Eloff JN. The anti-arthritic, anti-inflammatory, antioxidant activity and relationships with total phenolics and total flavonoids of nine South African plants used traditionally to treat arthritis. BMC Compl Altern Ned. 2016;16(1):1-10. https://doi.org/10.1186/s12906-016-1301-z

Khalid M, Alqarni MH, Shoaib A, Arif M, Foudah AI, Afzal O, Ali A, Ali A, Alqahtani SS, Altamimi AS. Anti-Arthritic and anti-inflammatory potential of Spondias mangifera extract fractions: an in silico, in vitro and in vivo approach. Plants. 2021;10(5):825. https://doi.org/10.3390/plants10050825

Joshi DG, Jat RK, Patil SB. In vitro protein denaturation and membrane stabilising anti-arthritic activity of aqueous extracts of bark of Ficus benghalensis L. against methotrexate. The Pharma Innovation Journal. 2021;10(4):689-92.

Marrassini C, Peralta I, Anesini C. Comparative study of the polyphenol content-related anti-inflammatory and antioxidant activities of two Urera aurantiaca specimens from different geographical areas. Chinese Med. 2018;13(1):1-2. https://doi.org/10.1186/s13020-018-0181-1

Derouich M, Bouhlali ED, Hmidani A, Bammou M, Bourkhis B, Sellam K, Alem C. Assessment of total polyphenols, flavonoids and anti-inflammatory potential of three Apiaceae species grown in the Southeast of Morocco. Sci Afr. 2020;9:e00507. https://doi.org/10.1016/j.sciaf.2020.e00507

Jose SM, Anilkumar M. In vitro anti inflammatory studies of the leaf extracts of Litsea quinqueflora (Dennst.) Suresh Biosci Biotechnol Res Commun. 2020;13(4):2022-28. http://dx.doi.org/10.21786/bbrc/13.4/58

Jose SM, Anilkumar M. Antibacterial and anti-inflammatory activity of the silver nano particles synthesised from the methanolic leaf extract of Litsea quinqueflora (Dennst.) Suresh. Adv Sci Eng Med. 2020;12(4):443-49. https://doi.org/10.1166/asem.2020.2551

Ozdal T, Capanoglu E, Altay F. A review on protein–phenolic interactions and associated changes. Food Res Int. 2013;51(2):954-70. https://doi.org/10.1016/j.foodres.2013.02.009

Kumarappan CT, Thilagam E, Mandal SC. Antioxidant activity of polyphenolic extracts of Ichnocarpus frutescens. Saudi J Biol Sci. 2012;19(3):349-55. https://doi.org/10.1016/j.sjbs.2012.04.004

Gangwar M, Gautam MK, Sharma AK, Tripathi YB, Goel RK, Nath G. Antioxidant capacity and radical scavenging effect of polyphenol rich Mallotus philippenensis fruit extract on human erythrocytes: an in vitro study. Sci World J. 2014. https://doi.org/10.1155/2014/279451

Choudhury D, Ghosal M, Das AP, Mandal P. In vitro antioxidant activity of methanolic leaves and barks extracts of four Litsea plants. Asian J Plant Sci Res. 2013;3:99-107.

Kunnaja P, Chansakaow S, Wittayapraparat A, Yusuk P, Sireeratawong S. In vitro antioxidant activity of Litsea martabanica root extract and its hepatoprotective effect on chlorpyrifos-induced toxicity in rats. Mol. 2021;26(7):1906. https://doi.org/10.3390/molecules26071906

Ambarwati NS, Elya B, Mahayasih PG, Awang MS, Omar H. Antioxidant activity of Litsea petiolata Hk. f. In Journal of Physics: Conference Series 2021;1869(1);012055. IOP Publishing. https://doi.org/10.1088/1742-6596/1869/1/012055

Published

21-08-2021 — Updated on 01-10-2021

How to Cite

1.
Jose SM, Anilkumar M. LCMS/MS analysis and evaluation of anti-inflammatory and antioxidant activities of the polyphenol fraction of Litsea quinqueflora (Dennst.) Suresh. Plant Sci. Today [Internet]. 2021 Oct. 1 [cited 2024 Dec. 22];8(4):865–872. Available from: https://horizonepublishing.com/journals/index.php/PST/article/view/1243

Issue

Section

Research Articles