Simultaneous Determination of Aegeline and Six Coumarins from Different Parts of the Plant Aegle marmelos Using UHPLC-PDA-MS and Chiral Separation of Aegeline Enantiomers Using HPLC-ToF-MS

Abstract

A fast UHPLC-PDA method was developed for the simultaneous analysis of one alkaloid, aegeline, and six coumarins, viz., umbelliferone, scopoletin, marmesinin, 8-hydroxypsoralen, angelicin, and marmelosin, from the leaf, fruit, root, and bark of Aegle marmelos. The UHPLC method was validated for linearity, accuracy, repeatability, limits of detection and limits of quantification. The linearity range (r² > 0.99) of the seven compounds was found to be 0.5–250 µg/mL, and the limits of detection and limits of quantification for the seven compounds were found to be 0.1 and 0.5 µg/mL, respectively. The developed UHPLC method is simple, fast, and especially suitable for quality control analysis of coumarins and aegeline from A. marmelos and commercial dietary supplements. Single quadrupole mass spectrometry was used for the identification and confirmation of coumarins and aegeline from different plant parts and dietary supplements. In addition, a novel chiral HPLC-ToF-MS method was developed for the resolution of aegeline enantiomers. By applying this chiral method, the distribution of enantiomers of aegeline from different parts of A. marmelos and aegeline-containing dietary supplements is reported for the first time.

Key words

Aegle marmelos - Rutaceae - aegeline - coumarins - UHPLC-PDA-MS - HPLC-ToF-MS - chiral separation

• References

• 1 Bose TK. Fruits of India, tropical and subtropical. Calcutta: Nayaprakash; 1985: 498-504
• 2 Kesari AN, Gupta RK, Singh SK, Diwakar S, Watal G. Hypoglycemic and antihyperglycemic activity of Aegle marmelos seed extract in normal and diabetic rats. J Ethnopharmacol 2006; 107: 374-379
• 3 Upadhya S, Shanbhag KK, Suneetha G, Balachandra Naidu M, Upadhya S. A study of hypoglycemic and antioxidant activity of Aegle marmelos in alloxan induced diabetic rats. Indian J Physiol Pharmacol 2004; 48: 476-480
• 4 Kamalakkannan N, Mainzen Prince PS. Antihyperlipidaemic effect of Aegle marmelos fruit extract in streptozotocin-induced diabetes in rats. J Sci Food Agr 2005; 85: 569-573
• 5 Singanan V, Singanan M, Begum H. The hepatoprotective effect of bael leaves (Aegle marmelos) in alcohol induced liver injury in albino rats. Int J Sci Techol 2007; 2: 83-92
• 6 Jagetia GC, Venkatesh P, Baliga MS. Aegle marmelos (L.) Correa inhibits the proliferation of transplanted Ehrlich ascites carcinoma in mice. Biol Pharm Bull 2005; 28: 58-64
• 7 Chakthong S, Weaaryee P, Puangphet P, Mahabusarakam W, Plodpai P, Voravuthikunchai SP, Kanjana-Opas A. Alkaloid and coumarins from the green fruits of Aegle marmelos . Phytochemistry 2012; 75: 108-113
• 8 Saha SK, Chatterjee A. Isolation of allo-imperatorin and β-sitosterol from the fruits of Aegle marmelos correa. J Indian Chem Soc 1957; 34: 228-230
• 9 Sharma BR, Rattan RK, Sharma P. Constituents of leaves and fruits of Aegle marmelos . Indian J Chem 1980; 19?B: 162
• 10 Sharma BR, Rattan RK, Sharma P. Constituents of Aegle marmelos. Part III. Marmelene, an alkaloid, and other components of unripe fruits of Aegle marmelos . Phytochemistry 1981; 20: 2606-2607
• 11 Sharma BR, Sharma P. Constituents of Aegle marmelos. II. Alkaloids and coumarin from fruits. Planta Med 1981; 43: 102-103
• 12 Bansal Y, Bansal G. Analytical methods for standardization of Aegle marmelos: A review. J Pharm Educ Res 2011; 2: 37-44
• 13 Central Council for Research in Ayurveda and Siddha. HPTLC-Fingerprint atlas of Ayurvedic single plant drugs mentioned in Ayurvedic Pharmacopoeia, Vol- III and IV-A. Report submitted to WHO, New Delhi, India: 3 – 4.
• 14 The Ayurvedic Pharmacopoeia of India, Part I, Vol. IV. New Delhi, India: Ministry of Health and Family Welfare, Department of Ayush; 2004: 12-13
• 15 The Ayurvedic Pharmacopoeia of India, Part I, Vol. III. New Delhi, India: Ministry of Health and Family Welfare, Department of Ayush; 2001: 29-31
• 16 Srivastava KK, Singh HK. Physico-chemical quality of bael (Aegle marmelos Correa) cultivars. Agric Sci Dig 2004; 24: 65-66
• 17 Dhankhar S, Ruhil S, Balhara M, Dhankhar S, Chhillar AK. Aegle marmelos (Linn.) Correa: A potential source of phytomedicine. J Med Plant Res 2011; 5: 1497-1507
• 18 Sharma N, Dubey W. History and taxonomy of Aegle marmelos: a review. Int J Pure App Biosci 2013; 1: 7-13
• 19 Vijayasankar R, Ravikumar K, Ravichandran P. Plant resources of Tiruvannamalai district, Tamil Nadu, India. Dehradun, India: Bishen Singh Mahendra Pal Singh; 2012: 171
• 20 Aegeline Identified as Harmful Ingredient in OxyElite Pro. Available at. http://www.huffingtonpost.com/2014/01/28/aegeline-oxyelite-pro_n_4683990.html Accessed on September 4, 2015
• 21 Nugroho AE, Riyanto S, Sukari MA, Maeyama K. Effects of aegeline, a main alkaloid of Aegle marmelos Correa leaves, on the histamine release from mast cells. Pak J Pharm Sci 2011; 24: 359-367
• 22 Manda VK, Avula B, Chittiboyina AG, Khan IA, Walker LA, Khan SI. Inhibition of CYP3A4 and CYP1A2 by Aegle marmelos and its constituents. Xenobiotica 2016; 46: 117-125
• 23 Gajbhiye NA, Makasana J, Thorat T. Simultaneous determination of marmin, skimmianine, umbelliferone, psoralene, and imperatorin in the root bark of Aegle marmelos by high-performance thin-layer chromatography. J Planar Chromatogr 2012; 25: 306-313
• 24 Shinde PB, Katekhaye SD, Mulik MB, Laddha KS. Rapid simultaneous determination of marmelosin, umbelliferone and scopoletin from Aegle marmelos fruit by RP-HPLC. J Food Sci Technol 2014; 51: 2251-2255
• 25 Bhattacherjee AK, Dikshit A, Pandey D, Tandon DK. High performance liquid chromatographic determination of marmelosin and psoralen in bael (Aegle marmelos (L.) Correa) fruit. J Food Sci Technol 2015; 52: 597-600
• 26 Karmase A, Prasanna K, Rasabattula S, Bhutani KK. Quantification and comparison of extraction methods for alkaloids in Aegle marmelos leaves by HPLC. Nat Prod Commun 2014; 9: 981-983
• 27 Shailajan S, Menon S, Singh A, Sayed N, Mhatre M. Marmelosin based standardization of Ayurvedic formulations containing Aegle marmelos using RP-HPLC method. J Pharm Res 2012; 5: 2224-2227
• 28 Sulaiman CT, George S, Balachandran I. Characterization of phenolic compounds in fruit extracts of three medicinal plants using liquid chromatography-negative electrospray ionization tandem mass spectrometry. Anal Chem Lett 2013; 3: 322-329
• 29 Dikshit BBL, Dutt S. Preliminary chemical examination of Aegle marmelos or the Indian Bel. J Indian Chem Soc 1930; 7: 759-764
• 30 Chopra RN. Indigenous drugs of India. 2nd. edition. Calcutta, India: Academic Publishers; 1933: 267-269
• 31 Faizi S, Farooqi F, Zikr-Ur-Rehman S, Naz A, Noor F, Ansari F, Ahmad A, Khan SA. Shahidine, a novel and highly labile oxazoline from Aegle marmelos: the parent compound of aegeline and related amides. Tetrahedron 2009; 65: 998-1004
• 32 Sadyandy R, Fernandes RA, Kumar P. An asymmetric dihydroxylation route to (R)-(−)-octopamine, (R)-(−)-tembamide and (R)-(−)-aegeline. ARKIVOC 2005; iii: 36-43
• 33 Albonico SM, Kuck AM, Deulofeu V. Tembamide from Fagara hiemalis . J Chem Soc C 1967; 1327-1328
• 34 Kamal A, Shaik AA, Sandbhor M, Malik MS. Chemoenzymatic synthesis of (R)- and (S)-tembamide, aegeline and denopamine by a one-pot lipase resolution protocol. Tetrahedron Asymmetry 2004; 15: 3939-3944
• 35 Patra A, Mitra AK, Ghosh A, Mukhopadhyay PK. Carbon-13 NMR spectra of tembamide, aegeline, and related amides. Org Magn Reson 1981; 16: 65-67
• 36 Somanathan R, Aguilar HR, Ventura GR, Smith KM. Syntheses of natural hydroxyamides using trimethylsilyl cyanide. Synth Commun 1983; 13: 273-280
• 37 ICH, Validation of Analytical Procedures: Text and Methodology, ICH Harmonized Tripartite Guidelines, November 2005. Available at http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf Accessed November 01, 2015