Detection and Estimation of alpha-Amyrin, beta-Sitosterol, Lupeol, and n-Triacontane in Two Medicinal Plants by High Performance Thin Layer Chromatography
A normal phase high performance thin layer chromatography (HPTLC) method has been developed and validated for simultaneous estimation of four components, namely, alpha-amyrin, beta-sitosterol, lupeol, and n-triacontane from two medicinally important plants, Leptadenia reticulata Wight & Arn. and Pluchea lanceolata (DC.) CB. Clarke. In Ayurveda, both plants have been reported to possess immunomodulatory activity. Chromatographic separation of the four components from the methanolic extracts of whole plant powders of Leptadenia reticulata Wight & Arn. and Pluchea lanceolata (DC.) CB. Clarke. was performed on TLC aluminium plates precoated with silica gel using a suitable mobile phase. The densitometric scanning was done after derivatization at λ = 580？nm for α-amyrin, β-sitosterol, and lupeol, and at 366？nm for n-triacontane. The developed HPTLC method has been validated and used for simultaneous quantitation of the four components from the methanolic extracts of whole plant powders of Leptadenia reticulata Wight & Arn. and Pluchea lanceolata (DC.) CB. Clarke. The developed HPTLC method is simple, rapid, and precise and can be used for routine quality control. 1. Introduction Herbal medicines have been used since ages to treat various ailments. Ayurveda is an Indian traditional system of medicine used since ancient times. It has a huge list of herbs used in various forms for treatment of different disease conditions. Owing to the medicinal properties attributed to herbal drugs, it is necessary to maintain their quality and purity, thereby justifying their acceptability in modern system of medicine. Standardisation of these herbal drugs is a challenge to the entire scientific fraternity. However, due to lack of suitable quality control and quality assurance standards for herbal drugs, it becomes difficult to ensure uniformity of their composition which in turn affects the efficacy of their final products. Analytical tools are important for qualitative, semiquantitative, and quantitative phytochemical analysis of herbal drugs and formulations. Chromatographic techniques such as high performance liquid chromatography (HPLC), high performance thin layer chromatography (HPTLC), and gas chromatography (GC) are used to efficiently determine the quality of the herbs by developing fingerprints and estimation of biomarkers. Among the wide choice of chromatographic techniques, HPTLC is a simple, fast, and accurate technique for use, making it advantageous over others for quick assessment of a number of samples simultaneously [1]. In the present research work, HPTLC
References
[1]
E. Reich and A. Schibli, High-Performance Thin-Layer Chromatography for the Analysis of Medicinal Plants, Thieme Medical, 2011.
[2]
S. Srivastav, D. Deepak, and A. Khare, “Three novel pregnane glycosides from Leptadenia reticulata Wight and Arn,” Tetrahedron, vol. 50, no. 3, pp. 789–798, 1994.
[3]
V. N. Dwivedi, Bhav Prakash Nighantu, Hindi Translational Motilal Banarsidas Banaras, 1949.
[4]
Anonymous, The Wealth of India Raw Materials Publication and Information Directorate, CSIR, New Delh, India, 1969.
[5]
H. Wagner, Immunomodulatory Agents from Plants, pp. 291-292, Birkh？user, Basel, Switzerland, 1st edition, 1998.
[6]
B. S. Kaith, “Neolupeol and anti-inflammatory activity of Pluchea lanceolata,” International Journal of Pharmacognosy, vol. 34, pp. 73–75, 1995.
[7]
M. Ali, N. Ali Siddiqui, and R. Ramachandram, “Phytochemical investigation of aerial parts of Pluchea lanceolata C.B. Clarke,” Indian Journal of Chemistry B, vol. 40, no. 8, pp. 698–706, 2001.
[8]
L. Fraile, E. Crisci, L. Córdoba, M. A. Navarro, J. Osada, and M. Montoya, “Immunomodulatory properties of beta-sitosterol in pig immune responses,” International Immunopharmacology, vol. 13, no. 3, pp. 316–321, 2012.
[9]
M. Martelanc, I. Vovk, and B. Simonovska, “Separation and identification of some common isomeric plant triterpenoids by thin-layer chromatography and high-performance liquid chromatography,” Journal of Chromatography A, vol. 1216, no. 38, pp. 6662–6670, 2009.
C. P. Khare, Indian Herbal Remedies: Rational Western Therapy, Ayurvedic , and Other Traditional Usage and Botany, Springer, 2004.
[12]
D. Mammen, M. Daniel, and R. T. Sane, “Seasonal and geographical variations in chemical constituents of Leptadenia reticulata,” International Journal of Pharmaceutical Sciences Review and Research, vol. 4, no. 2, pp. 111–116, 2010.
[13]
M. Martelanc, I. Vovk, and B. Simonovska, “Determination of three major triterpenoids in epicuticular wax of cabbage (Brassica oleracea L.) by high-performance liquid chromatography with UV and mass spectrometric detection,” Journal of Chromatography A, vol. 1164, no. 1-2, pp. 145–152, 2007.
[14]
T. A. Ibrahim, “Chemical composition and biological activity of extracts from Salvia bicolor desf. growing in Egypt,” Molecules, vol. 17, no. 10, pp. 11315–11334, 2012.
[15]
D. S. S. Kpoviéssi, F. Gbaguidi, J. Gbénou, et al., “Validation of a method for the determination of sterols and triterpenes in the aerial part of Justicia anselliana (Nees) T. Anders by capillary gas chromatography,” Journal of Pharmaceutical and Biomedical Analysis, vol. 48, no. 4, pp. 1127–1135, 2008.
[16]
E. H. Omoregie, K. F. Oluyemisi, O. S. Koma, and O. J. Ibumeh, “Chemical constituents of the essential oil of laggera pterodonta (DC.) sch. bip. from north-central Nigeria,” Journal of Applied Pharmaceutical Science, vol. 2, no. 8, pp. 198–202, 2012.
