LC-ESI-MS/MS Alkaloidal Profiling and Biological Investigation of Cleistocactus winteri Stems

Document Type : Research article


1 Department of Pharmacognosy, Faculty of Pharmacy, Helwan University, 1179, Cairo, Egypt.

2 National Research Centre, Department of Chemistry of Natural Compounds, Dokki, 12622, Giza, Egypt.


Cleistocactus winteri is a succulent plant of the Cactaceae family, commonly known as the golden rat tail. Many members of the Cactaceae family are the focus of chemical and biological studies as they contain bioactive compounds, well known for their health-related properties. We aimed to investigate Cleistocactus winteri stems both phytochemically and biologicallyfor the first timeincluding three biological effects. For the evaluation of theanti-inflammatory activity, Nitric oxide (NO) inhibition on lipopolysaccharide (LPS) stimulated Raw murine microphages (RAW 264.7) cell model was used. The cytotoxic activity was evaluated against hepatocarcinoma (HepG2), breast adenocarcinoma (MCF-7), colorectal carcinoma (HCT-116), and colon adenocarcinoma (CACO 2) human cancer cell lines using MTT (3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide) assay. The antioxidant activity was evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. Seven alkaloids were tentatively identified based on Mass bank on line library in addition to the isolation and identification of four known flavonoids and a β-sitosterol 3- O-glucoside from the studied extract. The stem methanol extract showed a 6% inhibition of NO production (up to 48 μmol/L). Furthermore, the IC50 of the total alkaloid fraction against HepG2, MCF-7, and CACO-2 cell lines was equal to 26.53, 23.8, and 13.07 µg/mL, respectively, while that of the methanol extract against Hep-G2 and HCT-116 was 181 µg/mL and 357 µg/mL, respectively with no effect against MCF-7 cell line. The antioxidant effect of the extract was about one third (112 µg/mL) that of ascorbic acid.

Graphical Abstract

LC-ESI-MS/MS Alkaloidal Profiling and Biological Investigation of Cleistocactus winteri Stems


  • Hunt DR, Taylor NP. The genera of Cactaceae: towards a new consensus. Bradleya (1986) 4: 65-78.
  • Kaur M, Kaur A and Sharma R. Pharmacological actions of Opuntia ficus indica: A Review. Appl. Pharmceut. Sci. (2012) 2 (7): 15-18.
  • Valente LMM, Paixão D,  Nascimento A, Santos PFP, Scheinvar LA,  Moura MRL, Tinoco LW,  Gomes LNF and Silva Antiradical activity, nutritional potential and flavonoids of the cladodes of Opuntia monacantha (Cactaceae). Food Chemistry (2010) 123 (4, 15): 1127–1131.
  • Benayada Z, Martinez-Villaluengaa C, Frias J, Gomez-Cordovesa C and Es-Safib M. Phenolic composition, antioxidant and anti-inflammatory activities of extracts from Moroccan Opuntia ficus-indica flowers obtained by different extraction methods. Industrial Crops and Products; (2014) 62: 412–420.
  • Chahdoura H, Barreira CM, Barros L, Santos-Buelga C, Isabel CFRF and Achour L. Phytochemical characterization and antioxidant activity of Opuntia microdasys (Lehm.) Pfeiff flowers in different stages of maturity. Journal of Functional Foods; (2014)9: 27–37.
  • Brahmi D, Ayed Y, Hfaiedh M, Ben Mansour H, Bouaziz C, Zourgui L and Bacha H. Protective effect of cactus cladode extract against cisplatin induced oxidative stress, genotoxicity and apoptosis in mice: combination with phytochemical composition. BMC Complement Altern Med.; (2012)12: 111.
  • Abdul Malek SN, Shin SK, Abdul Wahab N, and Yaacob H. Cytotoxic components of Pereskia bleo (Kunth) DC. (Cactaceae) leaves. Molecules; (2009)14: 1713-1724.
  • Kim H, Choi HK, Moon JY, Kim YS, Mosaddik A and Cho SK. Comparative antioxidant and antiproliferative activities of red and white pitayas and their correlation with flavonoid and polyphenol content. J. Food Sci.; (2011)76: 38-45.
  • Hassan F, El-Razek A and Hassan AA. Nutritional value and hypoglycemic effect of Prickly Cactus Pear (Opuntia Ficus-Indica) fruit juice in alloxan-induced diabetic rats. Australian Journal of Basic and Applied Sciences; (2012) 5(10): 356-377.
  • Kazama CC, Uchida DT, Canzi KN, Souza P, Crestani S, Junior AG and Junior Involvement of arginine-vasopressin in the diuretic and hypotensive effects of Pereskia grandifolia Haw. (Cactaceae). Journal of Ethnopharmacology; (2012)144, (1): 86–93.
  • Tanakaa LYA, Oliveiraa AJB, Goncalvesb JE, Ciprianic TR, Souzac LM, Marquesd CA, Wernerd MFP, Baggiod CH, Gorinc PAJ, Sassakic GL and Iacominic M. An arabinogalactan with anti-ulcer protective effects isolated from Cereus Peruvianus. Carbohydrate Polymers; (2010) 82:714–721.
  • Ncibi S, Othman MB, Akacha A, Krifi MN and Zorgui L. 2008. Opuntia ficus indica extract protects against chlorpyrifos-induced damage on mice liver. Food and Chemical Toxicology. (2008) 46: 797-802.
  • Guevara-Figueroa T, Jiménez-Islas H, Reyes-Escogido ML, Mortensen AG, Laursen BB,  Lin L, De León-Rodríguez A, Fomsgaard IS and Barba de la Rosa Proximate composition, phenolic acids, and flavonoids characterization of commercial and wild nopal (Opuntia spp.). J. Food Comp Anal. (2010) 23(6): 525–32.
  • Okazaki S, Kinoshita K, Ito S, Koyama K,Yuasa H and Takahashi K. Triterpenoid saponins from Echinopsis macrogona (Cactaceae). Phytochemistry. (2011) 72:136–46.
  • Abdul-Wahab IR, Guilhon CC, Fernandes PD and Boylan F. Anti-nociceptive activity of Pereskia bleo (Cactaceae) leaves extracts. J Ethnopharmacol. (2012) 144: 741-6.
  • Starha R, Chybidziurova A, Lacny Z. Alkaloids of the genus Turbinicarpus (Cactaceae). Biochem Syst Ecol. (1999) 27: 839-41.
  • Starha R, Constituents of Gymnocalycium riojense Fric ex H.Till & W.Till (Cactaceae). Biochemical Systematics and Ecology. (2002) 30: 365-366.
  • Ortiz CMF, Da´vila P and Portilla LBH. Alkaloids from Neobuxbaumia species (Cactaceae). Biochemical Systematics and Ecology. (2003) 31: 581–585.
  • Ghosal S and Srivastava RS. β Phenethylamine, tetrahydroisoquinoline and indole Alkaloids of Desmodium tiliaefoliumPhytochemistry. (1973) 12(1): 193–197.
  • Bruhn JG and Agurell S. Cactaceae alkaloids XVIII: Two new alkaloids from Coryphantha calipensis Bravo. Journal of Pharmaceutical Sciences. (1974) 63(4): 574–576.
  • Miller JM, Bohmt Flavonol and dihydroflavonol glycosides of Echinocereus triglochidiatus var. Gurneyi. Phytochemistry. (1982) 21(4): 951-2.
  • Jimenez-Aspee F, Quispe C, Soriano MC, Gonzalez JF, Hüneke E, Theoduloz C and Schmeda-Hirschmann Antioxidant activity and characterization of constituents in copao fruits (Eulychnia acida Phil., Cactaceae) by HPLC-DAD-MS/MS. Food Res Inter. (2014) 62: 286-98.
  • Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JK and Tannenbaum SR. Analysis of nitrate, nitrite and 15N in biological fluids. Anal Biochem.(1982) 126:131-136.
  • Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays, J. Immunol. Methods.(1983) 65: 55-63
  • Brand-Williams W, Cuvelier ME, Berset Use of a free radical method to evaluate antioxidant activity LWT - Food Science and Technology. (1995) 28 (1): 25-30
  • Brandt SD, Freeman S, Fleet IA, Mc-Gagh P and Alder Analytical chemistry of synthetic routes to psychoactive tryptamines Part II. Characterization of the Speeter and Anthony synthetic route to N,N-dialkylated tryptamines using GC-EI-ITMS, ESI-TQ-MS-MS and NMR. Analyst. (2005)130: 330-44.
  • Agrawal PK, Bansal MC. Flavonoid glycosides. In: Studies in organic chemistry 39, 13C NMR of flavonoids. Agrawal, P.K. (ed). New York, Elsevier. (1989) 283-364.
  • Markham KR, Ternai B, Stanley R, Geiger H and Mabry TJ. Carbon-13 NMR studies of flavonoids glycosides and their acylated derivatives. Tetrahedron. (1978) 34: 1389.
  • Mustafa KIG, Nurettin Y, Hasan BS and Hasan G. Flavonol glycosides from Consolida armeniaca. Turk J Chem. (2000) 24: 191-7.
  • Gulnur T, Merve M, Erdem Y and Mustafa A. Main flavonoids of Tilia argentea Ex DC leaves. Turk J Chem. (2004) 28: 745-749.
  • Zhang Y, Jiao J, Liu C, Wu X and Zhang Y. Isolation and purification of four flavone C-glycosides from antioxidant of bamboo leaves by macroporous resin column chromatography and preparative high-performance liquid chromatography. Food Chem. (2007) 107(3): 1326-36.
  • Wen P, Han H, Wang R, Wang N and Yao X. C-glycosylfavones and aromatic glycosides from Campylotropis hirtella, Asian J Trad Med. (2007) 2 (4):149-153.
  • Khatun M, Billah M, Abdul Quader M. Sterols and Sterol Glucoside from Phyllanthus Dhaka Univ. J. Sci. (2012) 60(1): 5-10.
  • Newman DJ, Cragg GM. Natural products as sources of new drugs over the last 25 years. J Nat Prod. (2007) 70(3): 461-7.
  • Dehbi F, Hasib A, Tilaoui M, El-Batal H, Zaki N, Ouatmane A, Jaouad A and Zyad A. Bioactive constituents, antioxidant activity and in vitro cancer cell cytotoxicity of Moroccan Prickly Pear ( Opuntia ficus indica L.) juices. J Nat Sci Res. (2013) 3(14): 12-20.
  • Lu J, Bao J, Chen X, Huang M and Wang Y. Alkaloids Isolated from Natural Herbs as the Anticancer Agents. Evidence-Based Complementary and Alternative Medicine, Hindawi Publishing Corporation, (2012)1-12.