• Theerada Taesotikul Department of Biomedicine and Health Informatics, Faculty of Pharmacy, Silpakorn University
  • Nudchanart Kitcharoen Department of Industrial Pharmacy, Faculty of Pharmacy, Silpakorn University, Sanamchandra Palace Campus, Nakhon Pathom
  • Chatchai Chinpaisal Department of Biomedicine and Health Informatics, Faculty of Pharmacy, Silpakorn University, Sanamchandra Palace Campus, Nakhon Pathom
  • Patamawan Phuagphong Department of Biomedicine and Health Informatics, Faculty of Pharmacy, Silpakorn University, Sanamchandra Palace Campus, Nakhon Pathom
  • Srisombat Nawanopparatsakul Department of Biomedicine and Health Informatics, Faculty of Pharmacy, Silpakorn University, Sanamchandra Palace Campus, Nakhon Pathom




Citrus hystrix; Pomacea canaliculata; phytochemical pesticide


The objective of this work was to screen, identify, and quantify the phytocompounds contained within Citrus hystrix peel extracts, in different extraction solvents, for molluscicidal activities. The peel of
C. hystrix was extracted through the process of maceration with methanol. The phytochemical compounds found within the crude methanolic extract were further isolated through fractional partition with various solvents including hexane, dichloromethane, and butanol. Using thin-layer chromatography, phytochemical screenings for triterpenes, steroids, phenolic compounds, and alkaloids were carried out for each extract/fraction. The stability of the extracts/fractions were studied under static storage and accelerated conditions. The findings revealed that methanol was a suitable solvent for primary extraction, due to the high number and yield of compounds found, including non-polar, medium polar and high polar substances. After fractional extraction, non-polar compounds retained within hexane layer whereas both polar and medium polar substances were detected in methanol extract, dichloromethane and butanol fractions. The extracts were stable in normal storage conditions. Thus, this investigation could be of use for those conducting further research into the pharmacological activities of compounds contained within C. hystrix peel.


Halwart M. The golden apple snail Pomacea canaliculata in Asian rice farming systems: present impact and future threat. Int J Pest Manage. 1994;40:199-206.

Joshi RC, San Martín R, Saez-Navarrete C, Alarcon J, Sainz J, Antolin MM, et al. Efficacy of quinoa (Chenopodium quinoa) saponins against golden apple snail (Pomacea canaliculata) in the Philippines under laboratory conditions. Crop Prot. 2008;27(3):553-7.

Husak V. Copper and copper-containing pesticides: Metabolism, toxicity and oxidative stress. J Vasyl Stefanyk

Precarpathian National University. 2015;2.

Poppenga RH, Oehme FW. Chapter 7 - Pesticide use and associated morbidity and mortality in veterinary medicine. In: Krieger R, editor. Hayes' Handbook of Pesticide Toxicology (Third Edition). New York: Academic Press; 2010. p. 285-301.

Sithithaworn P, Andrews R, Shekhovtsov SV, Mordvinov VA, Furman DP. Helminth-Trematode: Opisthorchis viverrini and Opisthorchis felineus. In: Motarjemi Y, editor. Encyclopedia of Food Safety. Waltham: Academic Press; 2014. p. 170-8.

Stoleru, V., Sellitto, VM. Pest Control in Organic Systems. In: Gill, H. K., Goyal, G., editors. Integrated Pest Management (IPM): Environmentally Sound Pest Management [Internet]. London: IntechOpen; 2016 [cited 2022 Aug 15]. Available from: https://www.intechopen.com/chapters/51826 doi: 10.5772/64457

Agarwal M, Verma A. Modern Technologies for Pest Control: A Review. In: Nazal MK, Zhao H, editors. Heavy Metals - Their Environmental Impacts and Mitigation [Internet]. London: IntechOpen; 2020 [cited 2022 Aug 15]. Available from: https://www.intechopen.com/chapters/73098 doi: 10.5772/ intechopen.93556

Aktar MW, Sengupta D, Chowdhury A. Impact of pesticides use in agriculture: their benefits and hazards. Interdiscip Toxicol. 2009;2(1):1-12.

Boudh S, Singh JS. Pesticide contamination: Environmental problems and remediation strategies. In: Bharagava RN, Chowdhary P, editors. Emerging and Eco-Friendly Approaches for Waste Management. Singapore: Springer Singapore; 2019. p. 245-69.

Booker CJ, Bedmutha R, Vogel T, Gloor A, Xu R, Ferrante L, et al. Experimental investigations into the insecticidal, fungicidal, and bactericidal properties of pyrolysis bio-oil from tobacco leaves using a fluidized bed pilot plant. Ind Eng Chem Res. 2010;49(20):10074-9.

Madreseh-Ghahfarokhi S, Pirali Y, Dehghani-Samani A, Dehghani-Samani A. The insecticidal and repellent activity of ginger (Zingiber officinale) and eucalyptus (Eucalyptus globulus) essential oils against Culex theileri Theobald, 1903 (Diptera: Culicidae). Ann Parasitol. 2018;64(4):351-60.

Ruttanaphan T, de Sousa G, Pengsook A, Pluempanupat W, Huditz H-I, Bullangpoti V, et al. A novel insecticidal molecule extracted from Alpinia galanga with potential to control the pest insect Spodoptera frugiperda. Insects. 2020;11:686.

Tofel KH, Kosma P, Stähler M, Adler C, Nukenine EN. Insecticidal products from Azadirachta indica and Plectranthus glandulosus growing in Cameroon for the protection of stored cowpea and maize against their major insect pests. Ind Crops Prod. 2017;110:58-64.

Li X, Deng Fe, Shan X, Pan J, Yu P, Mao Z. Effects of the molluscicidal agent GA-C13:0, a natural occurring ginkgolic acid, on snail mitochondria. Pestic Biochem Physiol. 2012;103(2):115-20.

Dandekar DV, Jayaprakasha GK, Patil BS. Hydrotropic extraction of bioactive limonin from sour orange (Citrus aurantium L.) seeds. Food Chem. 2008 Aug 1;109(3):515-20.

Xie F, Zhang M, Zhang CF, Wang ZT, Yu BY, Kou JP. Anti-inflammatory and analgesic activities of ethanolic extract and two limonoids from Melia toosendan fruit. J ethnopharmacol. 2008 May 22;117(3):463-6.

Dai L, Wang W, Dong X, Hu R, Nan X. Molluscicidal activity of cardiac glycosides from Nerium indicum against Pomacea canaliculata and its implications for the mechanisms of toxicity. Environ Toxicol Pharmacol. 2011 Sep 1;32(2):226-32.

Dai L, Qian X, Nan X, Zhang Y. Effect of cardiac glycosides from Nerium indicum on feeding rate, digestive enzymes activity and ultrastructural alterations of hepatopancreas in Pomacea canaliculata. Environ Toxicol Pharmacol. 2014 Jan 1;37(1):220-7.

Meiyanto E, Hermawan A, Anindyajati A. Natural products for cancer-targeted therapy: citrus flavonoids as potent chemopreventive agents. Asian Pac J Cancer Prev. 2012;13(2):427-36.

Nazari M, Ghorbani A, Hekmat-Doost A, Jeddi-Tehrani M, Zand H. Inactivation of nuclear factor-κB by citrus flavanone hesperidin contributes to apoptosis and chemo-sensitizing effect in Ramos cells. Eur J pharmacol. 2011;650(2-3):526-33.

Benavente-García O, Castillo J, Marin F, Ortuño A, Del Río J. Uses and properties of citrus flavonoids. J Agric Food Chem. 1997;45(12):4505-4515.

FAPA. Toxic effects of Citrus extracts on Pomacea Canaliculata. Proceeding of the 23rd Federation of Asian Pharmaceutical Associations Congress; 2010 Nov 5-8; Taipei, Taiwan. Taipei; 2010.

Sirichoti K, Pongpidjayamaad T, editors. Molluscicidal activity of Citrus hystrix extracts against Pomacea canaliculata. Proceeding of New Perspective in Multidisciplinary Research & Practice; 2015 July 9-11; Ho Chi Minh City, Vietnam. Bangkok: The Association of International Thailand; 2015.

Phuagphong P, Nawanopparatsakul S, Kitcharoen N, Chinpaisal C, Nguanboonmak J, Ayukarn N. Mechanisms of molluscicidal activity of Citrus hystrix against Pomacea canaliculata. Isan J Pharm Sci. 2015;10:53-60.

Harborne JB. Phytochemical methods. Chapman and Hall, London, 1998, p.60-66.

Nogata Y, Sakamoto K, Shiratsuchi H, Ishii T, Yano M, Ohta H. Flavonoid composition of fruit tissues of citrus species. Biosci Biotechnol Biochem. 2006;70(1):178-92.

Kuljarachanan T, Devahastin S, Chiewchan N. Evolution of antioxidant compounds in lime residues during drying. Food Chem. 2009;113:944-9.

Anchisi C, Maccioni A, Sinico C, Valenti D. Stability studies of new cosmetic formulations with vegetable extracts as functional agents. Il Farmaco. 2001;56(5-7):427-431.

Reich E. and Schibli A. High-performance thin-layer chromatography for the analysis of medicinal plants. Thieme Medical Publishers, New York, 2006, p.234.

Chinapongtitiwat V, Jongaroontaprangsee S, Chiewchan N, Devahastin S. Important flavonoids and limonin in selected Thai citrus residues. J Funct Foods. 2013;5(3):1151-58.






Original Research Articles