Chemical profiling of endangered Citrus macroptera leaf extracts and evaluation of its cytotoxic activity

Authors

  • Ritupriya Singh Applied Plant Biotechnology Research Lab, Centre for Plant and Environmental Biotechnology Amity Institute of Biotechnology, Amity University, Noida, Uttar Pradesh, 201 313, India https://orcid.org/0000-0003-4942-3189
  • Susmita Shukla Applied Plant Biotechnology Research Lab, Centre for Plant and Environmental Biotechnology Amity Institute of Biotechnology, Amity University, Noida, Uttar Pradesh, 201 313, India https://orcid.org/0000-0002-9172-3647
  • Shiv Kant Shukla School of Services, Kaushalya: The Skill University, Ahmedabad, Gujarat, 380 052, India https://orcid.org/0000-0002-7381-8984
  • Deepshikha Pande Katare Proteomics and Translational Research Lab, Centre for Medical Biotechnology, Amity Institute of Biotechnology, Amity University, Noida Uttar Pradesh, 201 313, India https://orcid.org/0000-0002-1881-3566

DOI:

https://doi.org/10.14719/pst.3044

Keywords:

Antioxidants, cytotoxicity, flavonoids, GC-MS, HPTLC, Phytoconstituents

Abstract

The main component of drug production and formulation, with minimal to no adverse effects, is plant-based medicine. Citrus macroptera is an endangered species of North-Eastern India and has a diverse array of pharmaceutical applications owing to the presence of several bioactive compounds. The study aimed to evaluate the presence of bioactive metabolites, antioxidants, and cytotoxic mechanisms of the in vitro and ex-vitro leaf extracts of C.macroptera. Gas chromatography–mass spectrometry (GC-MS) profiling was performed using the ethanolic leaf extracts. Methanolic and aqueous extracts were profiled using High-performance thin-layer chromatography (HPTLC). Moreover, the cytotoxicity of the methanolic leaf extracts was assessed using the 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) test against SH-SY5Y, human neuroblastoma cell lines. GC-MS analysis revealed the existence of numerous pharmacologically significant components such as esters, hydrocarbons, fatty alcohols, and ester compounds. The presence of flavonoids and stigmasterol was exhibited by the HPTLC profile. The in-situ HPTLC analysis revealed white fluorescence after derivatization under white light that explicitly confirms the antioxidant potential of the extracts and can serve as a lead source of natural antioxidants. The cytotoxic efficacy of the extracts was observed in a concentration-dependent manner. However, the in vitro leaf extract displayed better suppressive effects against the SH-SY5Y cells with a 24-h IC50 of 167.71?g/mL compared to the ex-vitro leaf extract. Further, a comprehensive study is necessary to identify the cytotoxic compounds and their mode of action. The outcome of our investigation provides empirical evidence that phytoconstituents present in the leaf extracts of C. macroptera can be used as an effective pharmaceutical agent to combat several diseases.

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References

Carvalho AR. Popular use, chemical composition, and trade of Cerrado's medicinal plants (Goiás, Brazil). Environ Dev Sustain. 2004;6:307-16. https://doi.org/10.1023/b:envi.0000029889.29698.89

Uprety Y, Asselin H, Boon EK, Yadav S, Shrestha KK. Indigenous use and bio-efficacy of medicinal plants in the Rasuwa district, Central Nepal. J Ethnobiol Ethnomed. 2010;6:1-10. https://doi.org/10.1186/1746-4269-6-3

Lv X, Zhao S, Ning Z, Zeng H, Shu Y, Tao O et al. Citrus fruits as a treasure trove of active natural metabolites that potentially provide benefits for human health. Chem Cent J. 2015;9:1-4. https://doi.org/10.1186/s13065-015-0145-9

Upadhaya A, Chaturvedi SS, Tiwari BK. Utilization of wild Citrus by Khasi and Garo tribe of Meghalaya. Indian J Tradit Knowl. 2016;15(1):121-27.

Malik SK, Uchoi A, Kumar S, Choudhary R, Pal D, Kole PR et al. Molecular characterization of Citrus macroptera Montr. (Satkara): An endangered wild species from North East India. Plant Biosyst. 2013;147:857-63. https://doi.org/10.1080/11263504.2012.751063

Gogoi M, Boruah JL, Bora PK, Das DJ, Famhawite V, Biswas A et al. Citrus macroptera induces apoptosis via death receptor and mitochondrial mediated pathway as prooxidant in human non-small cell lung cancer cells. Food Biosci. 2021;43:101293. https://doi.org/10.1016/j.fbio.2021.101293

Saini RK, Ranjit A, Sharma K, Prasad P, Shang X, Gowda KG, Keum YS. Bioactive compounds of citrus fruits: A review of composition and health benefits of carotenoids, flavonoids, limonoids and terpenes. Antioxidants. 2022;11:239. https://doi.org/10.3390/antiox11020239

Ekor M. The growing use of herbal medicines: Issues relating to adverse reactions and challenges in monitoring safety. Front Pharmacol. 2014;4:177. https://doi.org/10.3389/fphar.2013.00177

Sharma AK, Kumar R, Mishra A, Gupta R. Problems associated with clinical trials of Ayurvedic medicines. Rev Bras Farmacogn. 2010;20:276-81. https://doi.org/10.1590/s0102-695x2010000200023

De S, Nariya P. A new, rapid online-HPTLC method for evaluation of DPPH reduction with ascorbic acid. JPC-J Planar Chromat. 2013;26:21-25. https://doi.org/10.1556/jpc.26.2013.1.3

Hingu B, Sake S, Gupta H, Vyas N, Thakur KS, Thakur M. Study on the phytochemical, antioxidant and in vitro anticancer activity on root extract of Simarouba glauca DC. Plant Sci Today. 2023;10:98-105. https://doi.org/10.14719/pst.2481

Wallace WE, Moorthy AS. NIST mass spectrometry data center standard reference libraries and software tools: Application to seized drug analysis. J Forensic Sci. 2023. https://doi.org/10.1111/1556-4029.15284

Olufunmilayo EO, Gerke-Duncan MB, Holsinger RD. Oxidative stress and antioxidants in neurodegenerative disorders. Antioxidants. 2023;12:517. https://doi.org/10.3390/antiox12020517

Hussain G, Zhang L, Rasul A, Anwar H, Sohail MU, Razzaq A et al. Role of plant-derived flavonoids and their mechanism in attenuation of Alzheimer’s and Parkinson’s diseases: An update of recent data. Molecules. 2018;23:814 https://doi.org/10.3390/molecules23040814

Rakkimuthu R, Ananthi P, Sathishkumar P, Sowmiya D. Chemical profiling of fern Cheilosoria mysurensis (Wall. ex Hook.) Ching & Shing and its biological activity. Plant Sci Today. 2023;10(1):91-95. https://doi.org/10.14719/pst.1872

Li M, Qian M, Jiang Q, Tan B, Yin Y, Han X. Evidence of flavonoids on disease prevention. Antioxidants. 2023;12:527. https://doi.org/10.3390/antiox12020527

J Hashim F, Vichitphan S, Boonsiri P, Vichitphan K. Neuroprotective assessment of Moringa oleifera leaves extract against oxidative-stress-induced cytotoxicity in SHSY5Y neuroblastoma cells. Plants. 2021;10(5):889. https://doi.org/10.3390/plants10050889

Cacique AP, Barbosa ÉS, Pinho GP, Silvério FO. Maceration extraction conditions for determining the phenolic compounds and the antioxidant activity of Catharanthus roseus (L.) G. Don. Ciência e Agrotecnologia. 2020;44. https://doi.org/10.1590/1413-7054202044017420

Pozharitskaya ON, Ivanova SA, Shikov AN, Makarov VG. Separation and evaluation of free radical-scavenging activity of phenol components of Emblica officinalis extract by using an HPTLC–DPPH method. J Sep Sci. 2007;30:1250-54. https://doi.org/10.1002/jssc.200600532

Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65:55-63. https://doi.org/10.1016/0022-1759(83)90303-4

Swamy MK, Arumugam G, Kaur R, Ghasemzadeh A, Yusoff MM, Sinniah UR. GC-MS based metabolite profiling, antioxidant and antimicrobial properties of different solvent extracts of Malaysian Plectranthus amboinicus leaves. J Ethnobiol Ethnomedicine. 2017;2017. https://doi.org/10.1155/2017/1517683

Halberstein RA. Medicinal plants: Historical and cross-cultural usage patterns. Ann Epidemiol. 2005;15:686-99. https://doi.org/10.1016/j.annepidem.2005.02.004

Petrovska BB. Historical review of medicinal plants’ usage. Pharmacog Rev. 2012;6:1. https://doi.org/10.4103/0973-7847.95849

Foresti ML, Errazu A, Ferreira ML. Effect of several reaction parameters in the solvent-free ethyl oleate synthesis using Candida rugosa lipase immobilised on polypropylene. Biochem Eng. 2005;25:69-77. https://doi.org/10.1016/j.bej.2005.04.002

Ukiva M. Antitumor promoting effects and cytotoxic activity against human cancer liners of triterpene diols and triols from edible Chrysanthemum flowers. Cancer Lett. 2002;177:7-12. https://doi.org/10.1016/s0031-9422(00)80640-8

Katerere DR, Gray AI, Nash RJ, Waigh RD. Antimicrobial activity of pentacyclic triterpenes isolated from African Combretaceae. Phytochemistry. 2003;63:81-88. https://doi.org/10.1016/s0031-9422(02)00726-4

Jung YY, Hwang ST, Sethi G, Fan L, Arfuso F, Ahn KS. Potential anti-inflammatory and anti-cancer properties of farnesol. Molecules. 2018;23(11):2827. https://doi.org/10.3390/molecules23112827

Jahangir T, Khan TH, Prasad L, Sultana S. Farnesol prevents Fe-NTA-mediated renal oxidative stress and early tumour promotion markers in rats. Hum & Exp Toxicol. 2006;25:235-42. https://doi.org/10.1191/0960327106ht616oa

Chaudhary SC, Alam MS, Siddiqui MS, Athar M. Chemopreventive effect of farnesol on DMBA/TPA-induced skin tumorigenesis: Involvement of inflammation, Ras-ERK pathway and apoptosis. Life Sci. 2009;85:196-205. https://doi.org/10.1016/j.lfs.2009.05.008

Crisóstomo-Ayala KA, Sabater-Jara AB, Pérez Manriquez C, Ferreres F, Gil-Izquierdo Á, Pedreño MÁ et al. Comparative study of metabolomic profile and antioxidant content of adult and in vitro leaves of Aristotelia chilensis. Plants. 2021;11(1):37. https://doi.org/10.3390/plants11010037

Trendafilova A, Jadranin M, Gorgorov R, Stanilova M. Bioactive compounds in wild, in vitro obtained, ex vitro adapted and acclimated plants of Centaurea davidovii (Asteraceae). Natural Product Communications. 2015;10:1934578X1501000609. https://doi.org/10.1177/1934578X1501000609

Goodarzi M, Russell PJ, Vander Heyden Y. Similarity analyses of chromatographic herbal fingerprints: A review. Anal Chim Acta. 2013;804:16-28. https://doi. org/10.1016/j.aca.2013.09.017

Rocamora CR, Ramasamy K, Lim SM, Majeed ABA, Agatonovic-Kustrin S. HPTLC based approach for bioassay-guided evaluation of antidiabetic and neuroprotective effects of eight essential oils of the Lamiaceae family plants. J Pharm Biomed Anal. 2020;178:112909. https://doi.org/10.1016/j.jpba.2019.112909

Nonglang FP, Khale A, Wankhar W, Bhan S. Pharmacognostic evaluation of Eranthemum indicum extracts for its in-vitro antioxidant activity, acute toxicology and investigation of potent bioactive phytocompounds using HPTLC and GCMS. Beni-Suef University Journal of Basic and Applied Sciences. 2022;11:1-7. https://doi.org/10.1186/s43088-022-00311-2

Adin SN, Gupta I, Ahad A, Aqil M, Mujeeb M. A developed high-performance thin-layer chromatography method for the determination of baicalin in Oroxylum indicum L. and its antioxidant activity. JPC-J Planar Chromat. 2022;35(4):383-93. https://doi.org/10.1007/s00764-022-00182-4

Karim N, Sakib AH, Sarkar MY, Hossain MS, Ansari MA, Dhar R. Study on in-vitro cytotoxic and thrombolytic activity of methanolic extract of Citrus macroptera (Fruit). J Sci Res Adv. 2015;2(2):90-94.

Vermeersch KA, Wang L, McDonald JF, Styczynski MP. Distinct metabolic responses of an ovarian cancer stem cell line. BMC Systems Biology. 2014;8:1-4. https://doi.org/10.1186/s12918-014-0134-y

Gade S, Rajamanikyam M, Vadlapudi V, Nukala KM, Aluvala R, Giddigari C et al. Acetylcholinesterase inhibitory activity of stigmasterol and hexacosanol is responsible for larvicidal and repellent properties of Chromolaena odorata. Biochim Biophys Acta Gen Subj. 2017;1861(3):541-50. https://doi.org/10.1016/j.bbagen.2016.11.044

Ahmed A, Akbar S, Shah WA. Chemical composition and pharmacological potential of aromatic water from Salix caprea inflorescence. Chin J Integr Med. 2017;1-5. https://doi.org/10.1007/s11655-017-2781-5

Wiraswati HL, Fauziah N, Pradini GW, Kurnia D, Kodir RA, Berbudi A et al. Breynia cernua: Chemical profiling of volatile compounds in the stem extract and its antioxidant, antibacterial, antiplasmodial and anti-cancer activity in-vitro and in-silico. Metabolites. 2023;13:281. https://doi.org/10.3390/metabo13020281

Shobier AH, Ismail MM, Hassan SW. Variation in anti-inflammatory, anti-arthritic and antimicrobial activities of different extracts of common Egyptian seaweeds with an emphasis on their phytochemical and heavy metal contents. Biol Trace Elem Res. 2023;201:2071-87. https://doi.org/10.1007/s12011-022-03297-1

Cai S, Huang C, Ji B, Zhou F, Wise ML, Zhang D, Yang P. In vitro antioxidant activity and inhibitory effect, on oleic acid-induced hepatic steatosis, of fractions and subfractions from oat (Avena sativa L.) ethanol extract. Food Chem. 2011;124:900-05. https://doi.org/10.1016/j.foodchem.2010.07.017

Dailey OD, Wang X, Chen F, Huang G. Anti-cancer activity of branched-chain derivatives of oleic acid. Anticancer Res. 2011;31:3165-69.

Reddy LH, Couvreur P. Squalene: A natural triterpene for use in disease management and therapy. Adv Drug Deliv Rev. 2009;61:1412-26. https://doi.org/10.1016/j.addr.2009.09.005

Kim SK, Karadeniz F. Biological importance and applications of squalene and squalane. Adv Food Nutr Res. 2012;65:223-33. https://doi.org/10.1016/b978-0-12-416003-3.00014-7

Ciriminna R, Bongiorno D, Scurria A, Danzì C, Timpanaro G, Delisi R et al. Sicilian Opuntia ficus-indica seed oil: Fatty acid composition and bio-economical aspects. Eur J Lipid Sci Technol. 2017;119:1700232. https://doi.org/10.1002/ejlt.201700232

Published

26-02-2024 — Updated on 01-04-2024

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Singh R, Shukla S, Shukla SK, Pande Katare D. Chemical profiling of endangered Citrus macroptera leaf extracts and evaluation of its cytotoxic activity. Plant Sci. Today [Internet]. 2024 Apr. 1 [cited 2024 May 1];11(2). Available from: https://horizonepublishing.com/journals/index.php/PST/article/view/3044

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