Nat Prod Sci.  2017 Jun;23(2):75-83. 10.20307/nps.2017.23.2.75.

Microwave Assisted Extraction, Optimization using Central Composite Design, Quantitative Estimation of Arjunic Acid and Arjunolic Acid using HPTLC and Evaluation of Radical Scavenging Potential of Stem Bark of Terminalia arjuna

• Khatkar S
• Nanda A
• Ansari SH

• KMID: 2387055
• DOI: http://doi.org/10.20307/nps.2017.23.2.75

Abstract

The optimization and microwave assisted extraction of stem bark of Terminalia arjuna, quantitative estimation of the marker compounds arjunic acid and arjunolic acid using HPTLC and the evaluation of free radical scavenging activity has been performed in this study. The central composite design was used for optimization and the values of parameters for optimized batch of microwave assisted extraction were 1000W (Power), 3 minutes (Time) and 1/120 (Solid/solvent ratio). The solvent system to carry out the HPTLC was toluene: acetic acid: ethyl acetate (5: 5: 0.5) and quantitative estimation was done using standard equations obtained from the marker compounds. The in-vitro free radical scavenging activity was performed spectrophotometrically using ascorbic acid as standard. The value of estimated percentage yield of arjunic acid and arjunolic acid was 1.42% and 1.52% which upon experimentation was obtained as 1.38% and 1.51% respectively. The DPPH assay of the different batches of microwave assisted extraction and marker compounds taken suggested that the marker compounds arjunic acid and the arjunolic acid were responsible for the free radical scavenging activity as the batch having the maximum percentage yield of the marker compounds showed best free radical scavenging effect as compared to standard ascorbic acid. The ICâ‚…â‚€ value of the optimized batch was found to be 24.72 while that of the standard ascorbic acid was 29.83. Hence, the yield of arjunic acid and arjunolic acid has direct correlation with the free radical scavenging activity of stem bark extract of Terminalia arjuna and have potential to serve as active lead compounds for free radical scavenging activity.

Keyword

Arjunic acid; Arjunolic acid; Central composite design; Free radical scavenging activity; HPTLC; Microwave assisted extraction

MeSH Terms

Acetic Acid
Ascorbic Acid
Microwaves*
Terminalia*
Toluene
Acetic Acid
Ascorbic Acid
Toluene

Figure

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  Fig. 1. Response Surface Plots and Contour plots for the effect of different parameters on percentage yield of arjunic acid in Terminalia arjuna stem bark extracts.

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  Fig. 2. Response Surface Plots and Contour plots for the effect of selected parameters on percentage yield of arjunolic acid in Terminalia arjuna stem bark extracts.

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  Fig. 3. Standard peak of Arjunic Acid.

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  Fig. 4. Standard peak of Arjunolic Acid.

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  Fig. 5. HPTLC chromatogram for different concentrations of standard arjunic acid and arjunolic acid at 366 nm.

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  Fig. 6. Area under the peak diagram of different concentrations of standard arjunolic acid and arjunic acid.

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  Fig. 7. Area under the peak diagram of different extracts obtained for the marker estimation.

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  Fig. 8. Percentage yield of marker compounds obtained through HPTLC.

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  Fig. 9. IC50 values of the test batches, the marker compounds and standard ascorbic acid.

Reference

References

(1). Sasidharan S.., Chen Y.., Saravanan D.., Sundram K. M.., Latha L. Y.Afr. J. Tradit. Complement Altern. Med. 2011. 8:1–4.

(2). Rufino M. S. M.., Alves R. E.., de Brito E. S.., Pérez-Jiménez J.., Saura-Calixto F.., Mancini-Filho J.Food Chem. 2010. 121:996–1002.

(3). Bandaranayake W. M.Wetl. Ecol. Manag. 2002. 10:421–452.

(4). Vlachojannis J.., Magora F.., Chrubasik S.Phytother. Res. 2011. 25:1102–1104.

(5). Staba E. J.., Chung A. C.Phytochemistry. 1981. 20:2495–2498.

(6). Aslam J.., Mujib A.., Nasim S. A.., Sharma M. P.Sci. Hortic. 2009. 119:325–329.

(7). Xie P.., Chen S.., Liang Y. Z.., Wang X.., Tian R.., Upton R. J.Chromatogr. A. 2006. 1112:171–180.

(8). Pozharitskaya O. N.., Ivanova S. A.., Shikov A. N.., Makarov V. G. J.Sep. Sci. 2007. 30:1250–1254.

(9). Pirisi F. M.., Cabras P.., Cao C. F.., Migliorini M.., Muggelli M. J.Agric. Food Chem. 2000. 48:1191–1196.

(10). de la Torre-Carbot K.., Jauregui O.., Gimeno E.., Castellote A. I.., Lamuela-Raventós R. M.., López-Sabater M. C. J.Agric. Food Chem. 2005. 53:4331–4340.

(11). Daferera D. J.., Ziogas B. N.., Polissiou M. G. J.Agric. Food Chem. 2000. 48:2576–2581.

(12). Akowuah G. A.., Zhari I.., Norhayati I.., Mariam A. J.Food Compos. Anal. 2006. 19:118–126.

(13). Mallavadhani U. V.., Sahu G.., Muralidhar J.Pharm. Biol. 2002. 40:508–511.

(14). Khatoon S.., Srivastava M.., Rawat A.., Mehrotra S.JPC-J. Planar Chromatogr. Mod. TLC. 2005. 18:364–367.

(15). Kumar A.., Lakshman K.., Jayaveera K. N.., Tripathi S. M.., Satish K. V.Jordan J. Pharm. Sci. 2010. 3:63–68.

(16). Pereira C. A.., Yariwake J. H.., Lanças F. M.., Wauters J. N.., Tits M.., Angenot L.Phytochem. Anal. 2004. 15:241–248.

(17). Mehta M.., Kaur N.., Bhutani K. K.Phytochem. Anal. 2001. 12:91–95.

(18). Hawthorne S. B.., Grabanski C. B.., Martin E.., Miller D. J. J.Chromatogr. A. 2000. 892:421–433.

(19). De Castro M. D. L.., Garcýa-Ayuso L. E.Anal. Chim. Acta. 1998. 369:1–10.

(20). Huddleston J. G.., Willauer H. D.., Swatloski R. P.., Visser A. E.., Rogers R. D.Chem. Commun. 1998. 16:1765–1766.

(21). Khan M. K.., Abert-Vian M.., Fabiano-Tixier A. S.., Dangles O.., Chemat F.Food Chem. 2010. 119:851–858.

(22). Dai J.., Mumper R. J.Molecules. 2010. 15:7313–7352.

(23). Núñez Sellés A. J.., Vélez Castro H. T.., Agüero-Agüero J.., González-González J.., Naddeo F.., De Simone F.., Rastrelli L. J.Agric. Food Chem. 2002. 50:762–766.

(24). Pourmortazavi S. M.., Hajimirsadeghi S. S. J.Chromatogr. A. 2007. 1163:2–24.

(25). Routray W.., Orsat V.Food Bioprocess Technol. 2012. 5:409–424.

(26). Kaufmann B.., Christen P.Phytochem. Anal. 2002. 13:105–113.

(27). Lu Y.., Ma W.., Hu R.., Dai X.., Pan Y. J.Chromatogr. A. 2008. 1208:42–46.

(28). Spigno G.., De Faveri D. M. J.Food Eng. 2009. 93:210–217.

(29). Devgan M.., Nanda A.., Ansari S. H.Pak. J. Pharm. Sci. 2013. 26:973–976.

(30). Devgun M.., Nanda A. R. U. N.., Ansari S. H.Acta Pol. Pharm. 2012. 69:475–485.

(31). Devgun M.., Nanda A.., Ansari, S. H; Swamy S. K.Res. J. Pharm. Technol. 2010. 3:644–649.

(32). Valko M.., Jomova K.., Rhodes C. J.., Kuèa K.., Musílek K.Arch. Toxicol. 2016. 90:1–37.

(33). Przyklenk K.., Kloner R. A.Circ. Res. 1989. 64:86–96.

(34). Prasad K.., Kalra J.Am. Heart J. 1993. 125:958–973.

(35). Puthur J. T.South Indian J. Biol. Sci. 2016. 2:14–17.

(36). Galano A.., Mazzone G.., Alvarez-Diduk R.., Marino T.., Alvarez-Idaboy J. R.., Russo N.Annu. Rev. Food Sci. Technol. 2016. 7:335–352.

(37). Fang Y.., Luo Y.., Yu P. J.Renew. Sust. Energ. 2016. 8:033103.

(38). Choi S.., Kim J. M.., Shin G. H.., Jung T. D.., Oh J. W.., Choi S. H.., Cho B. Y.., Lee J. H.., Lee O. H.FASEB J. 2016. 30:1174–1175.

(39). Martinez-Valverde I.., Perago M. J.., Provan G.., Chesson A. J.Sci. Food Agr. 2002. 82:323–330.

(40). Duthie G. G.., Duthie S. J.., Kyle J. A.Nut. Res. Rev. 2000. 13:79–106.

(41). Halder S.., Bharal N.., Mediratta P. K.., Kaur I.., Sharma K. K.Indian. J. Exp. Biol. 2009. 47:577–583.

(42). Chaudhari, M. Mengi S.Phytother. Res.,. 2006. 20:799–805.

(43). Chatha S. A. S.., Hussain A. I.., Asad R.., Majeed M.., Aslam N. J.Food Process Technol. 2014. 5:298.

(44). Bachaya H. A.., Iqbal Z.., Khan M. N.., Jabbar A.., Gilani A. H.., Din I. U.Int. J. Agric. Biol. 2009. 11:273–278.

(45). Sultana B.., Anwar F.., Przybylski R.Food Chem. 2007. 104:1106–1114.

(46). Manna P.., Sinha M.., Sil P. C.Arch. Toxicol. 2008. 82:137–149.

(47). Elsherbiny N. M.., Eladl M. A.., Al-Gayyar M. M. H.Cytokine. 2016. 77:26–34.

(48). Sumitra M.., Manikandan P.., Kumar D. A.., Arutselvan N.., Balakrishna K.., Manohar B. M.., Puvanakrishnan R.Mol. Cell. Biochem. 2001. 224:135–142.

(49). Sun F. Y.., Chen X. P.., Wang J. H.., Qin H. L.., Yang S. R.., Du G. H.Am. J. Chin. Med. 2008. 36:197–207.

(50). Pharmacopoeia of India. Herbs and Herbal Monographs: Indian Pharmacopoeial Commission; Ministry of Health and Family Welfare: India,. 2010. 8–9.

(51). Bondet V.., Brand-Williams W.., Berset C.LWT-Food Sci. Tech. 1997. 30:609–615.

(52). Novilla A.., Nawawi A.., Sugihartina G.., Widowati W.Cukurova Med. J. 2014. 39:224–233.