Simultaneous estimation of three triterpenoids from Carissa carandas using validated high performance liquid chromatography
DOI:
https://doi.org/10.22377/ijgp.v6i3.269Abstract
Background: Carissa carandas L. (Apocynaceae), an important fruit of tropical and subtropical arid region, has a great potential for the development of a new phytotherapeutic medicine. Due to an increase in commercial products of this fruit in the global market, an extensive phytochemical comparison seems imperative to understand its internal quality. Aims and Objective: To develop and validate an HPLC-DAD method for simultaneous estimation of three triterpenoids from C. carandas. Materials and Methods: A simple, sensitive and rapid HPLC-DAD method was developed for simultaneous estimation of lupeol, stigmasterol, and b-sitosterol from C. carandas fruits. Isocratic elution at a flow rate of 1.0 mL/min was employed on C18 Cosmosil column with PDA detection at 210 nm using acetonitrile and ethanol. Different parts of C. carandas and its fruits collected from different geographical region were screened for lupeol, stigmasterol, and b-sitosterol contents. Results: Proposed method for simultaneous estimation of lupeol,stigmasterol and b-sitosterol was found highly specific, reliable, and sensitive which allowed good recovery of all analytes. It was
validated as per ICH guidelines and proved to have good linearity, precision, repeatability, and stability. It was efficiently applied to screen different samples of C. carandas providing a mean to assess the quality and authenticity. Conclusion: The HPLC method could be an effective technique for routine analysis of lupeol, stigmasterol, and b-sitosterol simultaneously and monitoring the quality of fruit and its available products in cosmetic and pharmaceutical companies.
Key words: Carissa carandas, simultaneous estimation, triterpenoids, validation
Downloads
References
Pagliarussi RS, Freitas LAP, Bastos JK. A quantitative method for
the analysis of xanthine alkaloids in Paullinia cupana (guarana) by
capillary column gas chromatography. J Sep Sci 2002;25:371-4.
Sharma A, Reddy GD, Kaushik A, Shanker K, Tiwari RK,
Mukherjee A, et al. Analgesic and Anti-inflammatory Activity of
Carissa carandas Linn fruits and Microstylis wallichii Lindl Tubers.
Nat Prod Sci 2007;13:6-10.
Rai R, Misra KK. Micropropagation of Karonda (Carissa carandas)
through shoot multiplication. Sci Hort 2005;103:227-32.
Sen S, Chakraborty R, De BT, Raghavendra GHG, Debnath S.
Analgesic and anti-inflammatory herbs: A potential source of
modern medicine. Int J Pharm Sci Res 2010;1:32-44.
Bhaskar VH, Balakrishnan N. Analgesic, anti-inflammatory and
antipyretic activities of Pergularia daemia and Carissa carandas. Daru
J Pharm Sci 2009;17:168-74.
Hegde K, Joshi AB. Hepatoprotective effect of Carissa carandas
Linn root extract against CCl4 and paracetamol induced hepatic
oxidative stress. Indian J Exp Biol 2009;47:660-7.
The Ayurvedic Pharmacopoeia of India (API). Part I, Vol. 2, 1st ed.
Government of India, Ministry of Health and Family Welfare. New
Delhi: The Controller of Publications Civil Lines; 2001. p. 78-9.
Wei H, Sun L, Tai Z, Gao S, Xu W, Chen W. A simple and sensitive
HPLC method for the simultaneous determination of eight
bioactive components and fingerprint analysis of Schisandra
sphenanthera. Anal Chim Acta 2010;662:97-104.
Shailajan S, Joshi H. Optimized Separation and quantification
of pharmacologically active markers Quercetin, Kaempferol,
β-sitosterol and Lupeol from Cuscuta reflexa Roxb. J Pharm Res
;4:1851-3.
Brufau G, Canela MA, Rafecas M. Photosterols: Physiologic and
metabolic aspects related to cholesterol lowering properties. Nutr
Res 2008;28:217-25.
Sukumar D, Arimboor R, Arumughan C. HPTLC fingerprinting
and quantification of lignans as marker in sesame oil and its
polyherbal formulations. J Pharm Biomed Anal 2008;47:795-801.
Kamboj A, Saluja AK. Isolation of stigmasterol and β-sitosterol
from petroleum ether extract of Aerial parts of Ageratum conyzoides
(asteraceae). Int J Pharm Pharm Sci 2011;3:94-6.
ICH.org [Homepage on internet]. Switzerland: International
Conference on Harmonization (ICH). Validation of analytical
procedures: Methodology [Q2 (R1)]; 1998. Available from: http://
www.ich.org. [Last cited on 2011 Jun 4].
Heyden YV, Nijhuis A, Smeyers-Verbeke J, Vandeginste BGM,
Massart DC. Guidance of robustness/ruggedness tests in method
validation. J Pharm Biomed Anal 2001;24:723-53.
Li Y, Wu T, Wang ZT, Guo C. Simultaneous determination of ursolic
acid and lupeol in leaves of Ilex cornuta from different harvest area
by reversed phase high performance liquid chromatography. Chin
Pharmaceut J 2009;44:784-6.
Da Costa PA, Ballus CA, Teixeira-Filho J, Godoy HT. Phytosterols
and tocopherols content of pulps and nuts of Brazilian fruits. Food
Res Int 2010;43:1603-6.
Sorenson WR, Sullivan D. Determination of campesterol,
stigmasterol, and beta-sitosterol in saw palmetto raw materials and
dietary supplements by gas chromatography: Single-laboratory
validation. J AOAC Int 2007;89:22-34.
Borkovcova J, Janouscova E, Drackova M, Janstova B, Vor L.
Determination of sterols in dairy products and vegetable fats by
HPLC and GC methods. Czech J Food Sci 2009;27:217-9.
Berdeux O, Dutta PC, Dobarganes MC, Sebedio JL. Analytical
methods for quantification of modified fatty acids and sterols
formed as a result of processing. Food Anal Methods 2009;2:30-40.
Lagarda MJ, Garcia-Llatas G, Farre R. Analysis of phytosterols in
foods. J Pharm Biomed Anal 2006;41:1486-96.
Martelan M, Vovk I, Simonovska B. Separation and identification
of some common isomeric plant triterpenoids by thin-layer
chromatography and high-performance liquid chromatography.
J Chrom A 2009;1216:6662-70.
Ye JC, Chang WC, Hsieh DJY, Hsiao MW. Extraction and analysis
of β-sitosterol in herbal medicines. J Med Plants Res 2010;4:522-7.
Sharma A, Tiwari RK, Kaushik A, Tyagi LK, Shankar K, Virmani T,
et al. Standardisation of Carissa carandas Linn: A drug used in Indian
system of medicine as per W.H.O. Guidelines. Cont J Pharm Sci
;1:9-14.
Malik SK, Chaudhury R, Dhariwal OP, Bhandari DC. Genetic
resources of tropical underutilized fruits in India. New Delhi:
National Bureau of Plant Genetic Resources (NBPGR); 2010. p. 47-53.
Ars-Grin.gov. Maryland: Germplasm Resources Information
Network - (GRIN). USDA, ARS, National Genetic Resources
Program. National Germplasm Resources Laboratory. Available
from: http://www.ars-grin.gov/cgi-bin/npgs/html/splist.pl?2090.
[Last cited on 2011 Jul 6].
