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Phytochemical Investigation with Assessment of Cytotoxicity and Antibacterial Activities of Chloroform Extract of the Leaves of Kalanchoe pinnata



S.K. Biswas, A. Chowdhury, S.Z. Raihan, M.A. Muhit, M.A. Akbar and R. Mowla
 
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ABSTRACT

Kalanchoe pinnata is used in traditional medicines for the treatment of several health problems in Bangladesh. The study was designed to investigate the cytotoxicity and antibacterial activities of the chloroform extract of the leaves of Kalanchoe pinnata. Qualitative phytochemical analysis was also performed for the identification of several phytochemical compounds such as alkaloids, glycosides, steroids, gums, flavonoids, saponins, reducing sugar, tannins and terpenoids. The study revealed that the plant extract contained all of the phytoconstituents except steroids, flavonoids and terpenoids. The agar disc diffusion technique was used to investigate the antibacterial activities of the plant extract at 500 μg disc-1 against eight bacterial strains. The plant extract showed very low antibacterial activities with zone of inhibition ranging from 0.33±0.02 to 1.67±0.02 mm, while the maximum antibacterial action was recorded against E. coli with zone of inhibition of 1.67±0.02 mm but no sensitivity was found against V. cholerae. The antibacterial activity of the plant extract was compared with the standard drug, Levofloxacin which was used at 10 μg disc-1. Brine shrimp lethality bioassay was also done to find out the cytotoxic activity of the plant sample. The chloroform extract of the plant showed lethality against the brine shrimp nauplii (LC50: 125.89 and LC90: 234.42 μg mL-1. Finally, it was concluded that the chloroform extract of the plant leaves possessed considerable cytotoxic activity with less antibacterial potentials.

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S.K. Biswas, A. Chowdhury, S.Z. Raihan, M.A. Muhit, M.A. Akbar and R. Mowla, 2012. Phytochemical Investigation with Assessment of Cytotoxicity and Antibacterial Activities of Chloroform Extract of the Leaves of Kalanchoe pinnata. American Journal of Plant Physiology, 7: 41-46.

DOI: 10.3923/ajpp.2012.41.46

URL: https://scialert.net/abstract/?doi=ajpp.2012.41.46
 
Received: November 04, 2011; Accepted: January 24, 2012; Published: February 22, 2012



INTRODUCTION

Medicinal plants were used as the principal health care agent before the discovery of modern medicines (Shafaei et al., 2011). The plants and phytochemical compounds are used in folk medicine for the treatment of different types of diseases (Pareta et al., 2011). The medicinal plant Kalanchoe pinnata Linn belonging to the family of Crassulaceae is found in Bangladesh and the local name of the plant is patharkuchi (Ghani, 2003). Several pharmacological potentials such as immunosuppressive (Rossi-Bergmann et al., 2006), anti-inflammatory and analgesic (Joseph et al., 2011), hepatoprotective (Yadav and Dixit, 2003) and antitumor promoting activities (Supratman et al., 2001) were reported. Inflammation of nephrons (Hartalka et al., 2007), hypertension (Lans, 2006) and allergic conditions (Cruz et al., 2008) can also be treated with the plant extract.

The ethanol extract of the leaves of Kalanchoe pinnata extracts possessed significant antibacterial activity (Biswas et al., 2011) and the petroleum ether and aqueous extracts of the plant showed cytotoxicity and antifungal potentials (Chowdhury et al., 2011). The present study was designed to find out the cytotoxic properties and antibacterial activities of the chloroform extract of the leaves of Kalanchoe pinnata. The preliminary phytochemical screening of the plant extract was also done.

MATERIALS AND METHODS

Collection and preparation of plant samples: The leaves of the medicinal plant were collected in December 2009 at day time from Ramchandrapur, Magura, Bangladesh. The undesirable materials were separated from leaves and after that, the leaves were air-dried for three weeks. The plant parts were ground into fine powder and kept in an air-tight container until use. About 200 g of powder of the plant material was soaked in 1000 mL of chloroform in a glass container which was sealed and stored for three weeks followed by continuous shaking. Finally, the crude mixture was filtered and the obtained filtrate was completely evaporated to give a gummy concentrate as crude chloroform extract.

Preliminary phytochemical screening: The freshly prepared chloroform extract of Kalanchoe pinnata was qualitatively tested for the presence of phytochemical compounds such as alkaloids, glycosides, steroids, gums, flavonoids, saponins, reducing sugar, tannins and terpenoids. The standard procedures as preliminary screening (Trease and Evans, 1989) were used to identify the presence of different phytoconstituents in the plant extract.

Test of antibacterial activity and cytotoxicity of the plant extract: The cytotoxic activity of the crude extract was investigated by brine shrimp lethality bioassay (Bauer et al., 1966) and agar disc diffusion method (Meyer et al., 1982) was used to determine the growth of inhibition of the bacterial strains.

Statistical analysis: Three replicates of each sample were used for statistical analysis and the results of the experiment were expressed as mean±standard deviation (SD).

RESULTS

Table 1 showed the results of phytochemical screening of the chloroform extract of the plant which revealed the presence of alkaloids, glycosides, gums, saponins, tannins and reducing sugars but the extract did not reveal the presence of steroids, flavonoids and terpenoids.

Cytotoxic activity: The results of investigation of cytotoxic activity (Table 2) showed that the chloroform extract of the plant possessed moderate level of general toxicity in the brine shrimp lethality bioassay (LC50: 125.89 and LC90: 234.42 μg mL-1).

Antibacterial activity: Antibacterial activity of the chloroform extract was investigated against eight bacterial strains. The results of antibacterial test showed that the crude extract showed very low level of antibacterial activity against B. subtilis, B. megaterium, S. aureus, E. coli, P. aeruginosa, S. typhi and S. dysenteriae with zone of inhibition between 0.33±0.02 to 1.67±0.02 mm.

Table 1: Results of phytochemical analysis of the chloroform extract of the leaves of Kalanchoe pinnata
Image for - Phytochemical Investigation with Assessment of Cytotoxicity and Antibacterial Activities of Chloroform Extract of the Leaves of Kalanchoe pinnata
+: Present and -: Absent

Table 2: Brine shrimp lethality bioassay of the chloroform extract of the leaves of Kalanchoe pinnata
Image for - Phytochemical Investigation with Assessment of Cytotoxicity and Antibacterial Activities of Chloroform Extract of the Leaves of Kalanchoe pinnata

Table 3: Screening for antimicrobial activity of the chloroform extract of the leaves of Kalanchoe pinnata
Image for - Phytochemical Investigation with Assessment of Cytotoxicity and Antibacterial Activities of Chloroform Extract of the Leaves of Kalanchoe pinnata
Data were represented as Mean±SD of triplicate determination, -: No inhibition

The highest zone of inhibition or antibacterial activity was found against E. coli (1.67±0.02 mm) while no antibacterial activity was found against V. cholerae. The zone of inhibition of the plant extract is shown in Table 3.

DISCUSSION

Alkaloids, glycosides, gums, saponins and reducing sugars were identified in the chloroform extract of the leaves of Kalanchoe pinnata while Chowdhury et al. (2011) reported the presence of alkaloid, glycoside, gums, saponins, reducing sugar and tannins in the petroleum ether extract of the leaves of Kalanchoe pinnata. They also reported that the aqueous extract of the same plant contained alkaloids, glycosides, steroids, saponins, tannins. On the other hand, the ethanol extract of the medicinal plant possessed alkaloids, glycosides, steroids, gums, flavonoids, saponins, reducing sugars and tannins (Biswas et al., 2011). The reducing sugars, tannins and glycosides present in the extract may be responsible for antibacterial activity and cardiac stimulatory potentials (Sofowora, 1993; Hagerman et al., 1998).

From literature review, it is evident that the phytochemical compounds play an important role in the therapeutic effects of the plants such as flavonoids for free radical scavenging potentials and antinociceptive activities (Zakaria et al., 2006). Alkaloids and tannins have also significant effect in inhibition of pain perception (Uche and Aprioku, 2008; Rahman et al., 2011; Ramprasath et al., 2006). Moreover, tannins could prevent infectious diseases (Agbafor et al., 2011).

The obtained results also showed the cytotoxic activity against brine shrimp nauplii with LC50: 125.89 and LC90: 234.42 μg mL-1. In comparison with the cytotoxic activities of the extract of petroleum ether (LC50: 25.12 and LC90: 177.83 μg mL-1) and aqueous extract (LC50: 25.12 and LC90: 173.78 μg mL-1) of the leaves of the plant reported by Chowdhury et al. (2011) and that of ethanol extract (LC50: 100 and LC90: 204.17 μg mL-1) by (Biswas et al., 2011), the chloroform extract possessed less cytotoxic potentials. In addition to cytotoxic properties, the chloroform extract showed very low antibacterial activities showing zone of inhibition ranging from 0.33±0.02 to 1.67±0.02 mm while significant antibacterial activity was found in the ethanol extract of the leaves of the plant with zone of inhibition between 6.0±0.35 to 8.2±0.22 mm (Biswas et al., 2011).

CONCLUSION

The results obtained from this study suggested that the chloroform extract of the leaves of Kalanchoe pinnata might contain anticancer and antibacterial compounds. However, more research work is needed to isolate the active chemical compounds with their chemical structures elucidation.

ACKNOWLEDGMENT

The authors are grateful to the authority of BGC Trust University for extending their cordial support to perform this research work.

REFERENCES

  1. Agbafor, K.N., E.I. Akubugwo, M.E. Ogbashi, P.M. Ajah and C.C. Ukwandu, 2011. Chemical and antimicrobial properties of leaf extracts of Zapoteca portoricensis. Res. J. Med. Plant, 5: 605-612.
    CrossRef  |  


  2. Hagerman, A.E., K.M. Riedl, G.A. Jones, K.N. Sovik, N.T. Ritchard, P.W. Hartzfeld and T.L. Riechel, 1998. High molecular weight plant polyphenolics (Tannins) as biological antioxidants. J. Agric. Food Chem., 46: 1887-1892.
    CrossRef  |  Direct Link  |  


  3. Bauer, A.W., W.M.M. Kirby, J.C. Sherris and M. Turck, 1966. Antibiotic susceptibility testing by a standardized single disk method. Am. J. Clin. Pathol., 45: 493-496.
    CrossRef  |  PubMed  |  Direct Link  |  


  4. Rossi-Bergmann, B., S.S. Costa, M.B.S. Borges, S.A. da Silva, G.R. Noleto, M.L.M. Souza and V.L.G. Moraes, 2006. Immunosuppressive effect of the aqueous extract of Kalanchoe pinnata in mice. Phytothera. Res., 82: 399-402.
    CrossRef  |  


  5. Biswas, S.K., A. Chowdhury, J. Das, U.K. Karmakar and M.C. Shill, 2011. Assessment of cytotoxicity and antibacterial activities of ethanolic extracts of Kalanchoe pinnata L. (Family: Crassulaceae) leaves and stems. Int. J. Pharm. Sci. Res., 2: 2605-2609.
    Direct Link  |  


  6. Chowdhury, A., S.K. Biswas, J. Das, U.K. Karmakar, M.C. Shill and N. Dutta, 2011. Investigation of cytotoxicity and antifungal activities of petroleum ether and aqueous extracts of leaves and stems of Kalanchoe pinnata L. (Crassulaceae). Asian J. Plant Sci., 10: 274-277.
    CrossRef  |  Direct Link  |  


  7. Cruz, E.A., S.A. Da-Silva, M.F. Muzitano, P.M. Silva, S.S. Costa and B. Rossi-Bergmann, 2008. Immunomodulatory pretreatment with Kalanchoe pinnata extract and its quercitrin flavonoid effectively protects mice against fatal anaphylactic shock. Int. Immunopharmacol., 8: 1616-1621.
    PubMed  |  


  8. Ghani, A., 2003. Medicinal Plants of Bangladesh. 2nd Edn., Asiatic Society of Bangladesh, Dhaka, Bangladesh


  9. Harlalka, G.V., C.R. Patil and M.R. Patil, 2007. Protective effect of Kalanchoe pinnata pers. (Crassulaceae) on gentamicin-induced nephrotoxicity in rats. Indian J. Pharmacol., 39: 201-205.
    CrossRef  |  Direct Link  |  


  10. Joseph, B., S. Sridhar, Sankarganesh, Justinraj and B.T. Edwin, 2011. Rare medicinal plant-kalanchoe pinnata. Res. J. Microbiol., 6: 322-327.
    CrossRef  |  Direct Link  |  


  11. Lans, C.A., 2006. Ethnomedicines used in trinada and tabago for urinary for urinary problems and diabetes mellitus. J. Ethnobiol. Ethnomed., Vol. 2,
    CrossRef  |  Direct Link  |  


  12. Meyer, B.N., N.R. Ferrigni, J.E. Putnam, L.B. Jacobsen, D.E. Nichols and J.L. Mclaughlin, 1982. Brine shrimp: A convenient general bioassay for active plant constituents. Planta Med., 45: 31-34.
    CrossRef  |  PubMed  |  Direct Link  |  


  13. Pareta, S.K., K.C. Patra, P.M. Mazumder and D. Sasmal, 2011. Establishing the principle of herbal therapy for antiurolithiatic activity: A review. J. Pharmacol. Toxicol., 6: 321-332.
    CrossRef  |  Direct Link  |  


  14. Rahman, M.A., E. Haque, M. Hasanuzzaman and I.Z. Shahid, 2011. Antinociceptive, antiinflammatory and antibacterial properties of Tamarix indica roots. Int. J. Pharmacol., 7: 527-531.
    CrossRef  |  


  15. Shafaei, A., E. Farsi, B.M.K. Ahamed, M.J.A. Siddiqui, I.H. Attitalla, I. Zhari and M.Z. Asmawi, 2011. Evaluation of toxicological and standardization parameters and phytochemical investigation of Ficus deltoidea leaves. Am. J. Biochem. Mol. Biol., 1: 237-243.
    CrossRef  |  Direct Link  |  


  16. Sofowora, A., 1993. Medicinal Plant and Traditional Medicine in Africa. 2nd Edn., John Wiley and Sons, New York., USA., pp: 6-65


  17. Supratman, U., T. Fujita, K. Akiyama, H. Hayashi and A. Murkami et al., 2001. Anti-tumor promoting activity of bufadienolides from Kalanchoe pinnata and K. daigremontiana X tubiflora. Biosci. Biotechnol. Biochem., 65: 947-949.
    PubMed  |  Direct Link  |  


  18. Trease, G.E. and W.C. Evans, 1989. Trease and Evan's Textbook of Pharmacognosy. 13th Edn., Cambridge University Press, London, Pages: 546


  19. Uche, F.I. and J.S. Aprioku, 2008. The phytochemical constituents, analgesic and anti-inflammatory effects of methanol extract of Jatropha curcas sp leaves in mice and Wister albino rats. J. Applied Sci. Environ. Manage., 12: 99-102.
    Direct Link  |  


  20. Ramprasath, V.R., P. Shanthi and P. Sachdanandam, 2006. Immunomodulatory and anti-inflammatory effects of Semecarpus anacardium Linn. Nut milk extract in experimental inflammatory conditions. Biol. Pharm. Bull., 29: 693-700.
    CrossRef  |  Direct Link  |  


  21. Yadav, N.P. and V.K. Dixit, 2003. Hepatoprotective activity of leaves of Kalanchoe pinnata Pers. J. Ethnopharmacol., 86: 197-202.
    CrossRef  |  PubMed  |  Direct Link  |  


  22. Zakaria, Z.A., R.N.S.R.M. Nor, M.R. Sulaiman, Z.D.F.A. Ghani, G.H. Kumar and C.A. Fatimah, 2006. Antinociceptive and anti-inflammatory properties of Melastoma malabathricum leaves chloroform extract in experimental animals. J. Pharmacol. Toxicol., 1: 337-345.
    CrossRef  |  Direct Link  |  


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