Development and Validation of a Combined Methodology for Assessing the Total Quality Control of Herbal Medicinal Products – Application to Oleuropein Preparations
Oleuropein (OE) is a secoiridoid glycoside, which occurs mostly in the Oleaceae family presenting several pharmacological properties, including antioxidant, cardio-protective, anti-atherogenic effects etc. Based on these findings OE is commercially available, as Herbal Medicinal Product (HMP), claimed for its antioxidant effects. As there are general provisions of the medicine regulating bodies e.g. European Medicines Agency, the quality of the HMP’s must always be demonstrated. Therefore, a novel LC-MS methodology was developed and validated for the simultaneous quantification of OE and its main degradation product, hydroxytyrosol (HT), for the relevant OE claimed HMP’s. The internal standard (IS) methodology was employed and separation of OE, HT and IS was achieved on a C18 Fused Core column with 3.1 min overall run time employing the SIM method for the analytical signal acquisition. The method was validated according to the International Conference on Harmonisation requirements and the results show adequate linearity (r2 > 0.99) over a wide concentration range [0.1–15 μg/mL (n=12)] and a LLOQ value of 0.1 μg/mL, for both OE and HT. Furthermore, as it would be beneficial to control the quality taking into account all the substances of the OE claimed HMP’s; a metabolomics-like approach has been developed and applied for the total quality control of the different preparations employing UHPLC-HRMS-multivariate analysis (MVA). Four OE-claimed commercial HMP’s have been randomly selected and MVA similarity-based measurements were performed. The results showed that the examined samples could also be differentiated as evidenced according to their scores plot. Batch to batch reproducibility between the samples of the same brand has also been determined and found to be acceptable. Overall, the developed combined methodology has been found to be an efficient tool for the monitoring of the HMP’s total quality. Only one OE HMP has been found to be consistent to its label claim.
References
[1]
Agalias A, Melliou E, Magiatis P, Tsarbopoulos A, Mitaku S (2005) Quantitation of polyphenols and secoiridoids in decoctions of Olea europaea leaves from ten Greek cultivated varieties. J Liq Chromatogr 28: 1557–1571. doi:10.1081/JLC-200058355.
[2]
Romani N, Mulinacci P, Pinelli F, Vincieri FF, Cimato A (1999) Polyphenolic content in five tuscany cultivars of Olea europaea L. J Agric Food Chem 47: 964-967. doi:10.1021/jf980264t. PubMed: 10552399.
[3]
Japon-Lujan R, Luque de Castro MD (2006) Superheated liquid extraction of oleuropein and related biophenols from olive leaves. J Chromatogr A 1136: 185–191.
[4]
Renaud S, De Lorgeril M, Delaye J, Guidollet J, Jacquard F et al. (1995) Cretan Mediterranean diet for prevention of coronary heart disease. Am J Clin Nutr 6: 1360–1367. PubMed: 7754988.
[5]
Galli C, Visioli F (1999) Antioxidant and other activities of phenolics in olives/olive oil, typical components of the mediterranean diet. Lipids 34: 23-26. doi:10.1007/BF02562224. PubMed: 10188593.
[6]
Kontogianni VG, Gerothanassis IP (2012) Phenolic compounds and antioxidant activity of olive leaf extracts. Nat Prod Res 26: 186–189. doi:10.1080/14786419.2011.582842. PubMed: 22060136.
[7]
Pereira AP, Ferreira ICFR, Marcelino F, Valentao P, Andrade PB et al. (2007) Phenolic compounds and antimicrobial activity of olive (Olea europaea L. Cv. Cobrancosa) leaves. Molecules 12 (5): 1153–1162.
[8]
Omar SH (2010) Cardioprotective and neuroprotective roles of oleuropein in olive. 18: 111–121. doi:10.1016/j.jsps.2010.05.005. PubMed: 23964170.
[9]
Keys A (1997) Coronary heart disease in seven countries.1970. Nutrition 13(3): 249-253. doi:10.1016/S0899-9007(97)91292-2. PubMed: 9131697.
[10]
Andreadou I, Iliodromitis EK, Mikros E, Constantinou M, Agalias A et al. (2006) The olive constituent oleuropein exhibits anti-ischemic, antioxidative, and hypolipidemic effects in anesthetized rabbits. J Nutr 136: 2213–2219. PubMed: 16857843.
[11]
Visioli F, Galli C (2001) Antiatherogenic components of olive oil. Curr Atheroscler Rep 3: 64–67. doi:10.1007/s11883-001-0012-0. PubMed: 11123850.
[12]
Scoditti E, Calabriso N, Massaro M, Pellegrino M, Storelli C et al. (2012) Mediterranean diet polyphenols reduce inflammatory angiogenesis through MMP-9 and COX-2 inhibition in human vascular endothelial cells: A potentially protective mechanism in atherosclerotic vascular disease and cancer. Arch Biochem Biophys 527: 81-89. doi:10.1016/j.abb.2012.05.003. PubMed: 22595400.
[13]
Poudyal H, Campbell F, Brown L (2010) Olive Leaf Extract Attenuates Cardiac, Hepatic, and Metabolic Changes in High Carbohydrate High Fat Fed Rats. J Nutr 140: 946–953. doi:10.3945/jn.109.117812. PubMed: 20335636.
[14]
Jemai H, El Feki A, Sayadi S (2009) Antidiabetic and Antioxidant Effects of Hydroxytyrosol and Oleuropein from Olive Leaves in Alloxan Diabetic Rats. J Agric Food Chem 57(19): 8798-8804. doi:10.1021/jf901280r. PubMed: 19725535.
[15]
Acquaviva R, Di Giacomo C, Sorrenti V, Galvano F, Santangelo R et al. (2012) Antiproliferative Effect Of Oleuropein In Prostate Cell Lines. Int J Oncol 41(1): 31-38. PubMed: 22484302.
[16]
European Medicines Agency (2011) Assessment Report on Olea europaea L. , folium. Based on Article 16d(1), Article 16f and Article 16h of Directive 2001/83/EC as amended (traditional use). Assessment report on Olea europaea L., folium, EMA/HMPC/430506/2009. EMA website. Available: . http://www.ema.europa.eu/docs/en_GB/docu？ment_library/Herbal_-_HMPC_assessment_re？port/2012/04/WC500125459.pdf . Accessed 2013 Oct 3.
[17]
Granados-Principal S, Quiles JL, Ramirez-Tortosa CL, Sanchez-Rovira P, Ramirez-Tortosa MC (2010) Hydroxytyrosol: from laboratory investigations to future clinical trials. Nutr Rev 68(4): 191-206. doi:10.1111/j.1753-4887.2010.00278.x. PubMed: 20416016.
[18]
(2004) Directive 2004/24/EC of the European Parliament and of the Council, amending, as regards traditional herbal medicinal products, Directive 2001/83/EC on the Community code relating to medicinal products for human use. Official Journal of the European Union L 136-85. Available: http://eur-lex.europa.eu/LexUriServ/LexU？riServ.do?uri=OJ:L:2004:136:0085:0090:en？:PDF.
[19]
EMA website. Available: . http://www.ema.europa.eu/ema/index.jsp?c？url=pages/regulation/general/general_con？tent_000208.jsp&mid=WC0b01ac05800240cf . Accessed 2013 Oct 3.
[20]
Williamson EM et al. (2001) Synergy and other interactions in phytomedicines.Phytomedicine 5: 401-409. PubMed: 11695885.
[21]
Dierkes G, Krieger S, Dück R, Bongartz A, Schmitz OJ et al. (2012) High-Performance Liquid Chromatography？Mass Spectrometry Profiling of Phenolic Compounds for Evaluation of Olive Oil Bitterness and Pungency. J Agric Food Chem 60: 7597？7606. doi:10.1021/jf3020574. PubMed: 22804690.
[22]
Zoidou E, Melliou E, Gikas E, Tsarbopoulos A, Magiatis P et al. (2010) Identification of Throuba Thassos, a Traditional Greek Table Olive Variety, as a Nutritional Rich Source of Oleuropein. J Agric Food Chem 58: 46–50.
[23]
Sanchez JL, Giambanelli E, Quirantes-Piné R, Cerretani L et al. (2011) Wastes Generated during the Storage of Extra Virgin Olive Oil as a Natural Source of Phenolic Compounds. J Agric Food Chem 59: 11491–11500. doi:10.1021/jf202596q. PubMed: 21939275.
[24]
Bazoti FN, Gikas E, Tsarbopoulos A (2010) Simultaneous quantification of oleuropein and its metabolites in rat plasma by liquid chromatography electrospray ionization tandem mass spectrometry 24. Biomed Chromatogr. pp. 506–515.
[25]
Ortega-García F, Peragón J (2010) HPLC analysis of oleuropein, hydroxytyrosol, and tyrosol in stems and roots of Olea europaea L. cv. Picual during ripening. J Sci Food Agric 90: 2295–2300. doi:10.1002/jsfa.4085. PubMed: 20648529.
[26]
Tan HW, Tuck KL, Stupans I, Hayball PJ (2003) Simultaneous determination of oleuropein and hydroxytyrosol in rat plasma using liquid chromatography with fluorescence detection. 785: 187–191 doi:10.1016/S1570-0232(02)00855-3. PubMed: 12535851.
[27]
Zhou T, Qian T, Wang X, Li X, Cao L et al. (2011) Application of LC-MS/MS method for the in vivo metabolite determination of oleuropein after intravenous administration to rat. Biomed - Journal of Chromatogr 25(12): 1360-1363. doi:10.1002/bmc.1609. PubMed: 21308708.
[28]
Briante R, Patumi M, Terenziani S, Bismuto E, Febbraio F et al. (2002) Olea Europaea L. Leaf Extract And Derivatives: Antioxidant Properties. J Agric Food Chem 50: 4934？4940. doi:10.1021/jf025540p. PubMed: 12166985.
[29]
Aturki Z, Fanali S, D’Orazio G, Rocco A, Rosati C (2008) Analysis of phenolic compounds in extra virgin olive oil by using reversed-phase capillary electrochromatography. Electrophoresis 29: 1643–1650. doi:10.1002/elps.200700547. PubMed: 18383030.
[30]
Aouidi F, Dupuy N, Artaud J, Roussos S, Msallem M et al. (2012) Rapid quantitative determination of oleuropein in olive leaves (Olea europaea) using mid-infrared spectroscopy combined with chemometric analyses. Industrial Crops and Products 37: 292–297. doi:10.1016/j.indcrop.2011.12.024.
[31]
International Conference On Harmonisation Of Technical Requirements For Registration Of Pharmaceuticals For Human Use (2010) ICH Harmonised Tripartite Guideline Validation Of Analytical Procedures: Text And Methodology Q2 (R1). Current Step 4 version Parent Guideline dated 27 October 1994 (Complementary Guideline on Methodology dated 6 November 1996 incorporated in November 2005). ICH website. Available: http://www.ich.org/fileadmin/Public_Web_？Site/ICH_Products/Guidelines/Quality/Q2_？R1/Step4/Q2_R1__Guideline.pdf . Accessed 2013 Oct 3.
[32]
Yang P, Litwinski GR, Pursch M, McCabe T, Kuppannan K (2009) Separation of natural product using columns packed with Fused-Core particles. J Sep Sci 32: 1816–1822.
[33]
Manchón N, D’Arrigo M, García-Lafuente A, Guillamón E, Villares A et al. (2010) Fast analysis of isoflavones by high-performance liquid chromatography using a column packed with fused-core particles. Talanta 82(5): 1986–1994. doi:10.1016/j.talanta.2010.08.050. PubMed: 20875606.
[34]
Orozco-Solano MI, Ferreiro-Vera C, Priego-Capote F, de Castro Luque (2012) Automated method for determination of olive oil phenols and metabolites in human plasma and application in intervention studies. J Chromatogr A 1258: 108–116. doi:10.1016/j.chroma.2012.08.057. PubMed: 22944382.
[35]
Hadad GM, Abdel Salam RA, Emara S (2012) Validated and optimized high-performance liquid chromatographic determination of tizoxanide, the main active metabolite of nitazoxanide in human urine, plasma and breast milk. J Chromatogr Sci 50(6): 509-515. doi:10.1093/chromsci/bms041. PubMed: 22525879.
[36]
Lan K, Zhang Y, Yang J, Xu L (2010) Simple quality assessment approach for herbal extracts using high performance liquid chromatography-UV based metabolomics platform. J Chromatogr A 1217: 1414–1418. doi:10.1016/j.chroma.2009.12.031. PubMed: 20045112.
[37]
Zhang HM, Li SL, Zhang H, Wang Y, Zhao ZL et al. (2012) Holistic quality evaluation of commercial white and red ginseng using a UPLC-QTOF-MS/MS-based metabolomics approach. J Pharm Biomed Anal 62: 258–273. doi:10.1016/j.jpba.2012.01.010. PubMed: 22310552.
[38]
Agalias A, Magiatis P, Skaltsounis AL, Mikros E, Tsarbopoulos A et al. (2007) A New Process For The Management Of Olive Oil Mill Waste Water And Recovery Of Natural Antioxidants. J Agric Food Chem 55: 2671？2676. doi:10.1021/jf063091d. PubMed: 17348673.
[39]
Stagos D, Portesis N, Spanou C, Mossialos D, Aligiannis N et al. (2012) Correlation of total polyphenolic content with antioxidant and antibacterial activity of 24 extracts from Greek domestic Lamiaceae species. Food Chem Toxicol 50(11): 4115–4124. doi:10.1016/j.fct.2012.08.033. PubMed: 22939934.
