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Oil palm vegetation liquor

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Oil palm vegetation liquor: a new source of phenolic bioactivesRavigadevi Sambanthamurthi1*, YewAi Tan1, Kalyana Sundram2, Mahinda Abeywardena3,T. G. Sambandan4, ChoKyun Rha4, Anthony J. Sinskey4, Krishnan Subramaniam5, Soon-Sen Leow1,Kenneth C. Hayes6and Mohd Basri Wahid11Malaysian Palm Oil Board, 6, Persiaran Institusi, Bandar Baru Bangi, 43000 Kajang Selangor, Malaysia2Malaysian Palm Oil Council, 2nd Floor, Wisma Sawit, Lot 6, SS6, Jalan Perbandaran, 47301 Kelana Jaya,Selangor, Malaysia3Commonwealth Scientific and Industrial Research Organisation, Gate 13, Kintore Avenue, Adelaide, SA 5000, Australia4Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA5MAHSA University College, Jalan University Campus, 59100 Kuala Lumpur, Malaysia6Brandeis University, 415 South Street, Waltham, MA 02454, USA(Received 14 October 2010 – Revised 31 January 2011 – Accepted 3 March 2011)AbstractWaste from agricultural products represents a disposal liability, which needs to be addressed. Palm oil is the most widely traded edible oilglobally, and its production generates 85 million tons of aqueous by-products annually. This aqueous stream is rich in phenolic antioxi-dants, which were investigated for their composition and potential in vitro biological activity. We have identified three isomers of caffeoyl-shikimic acid as major components of oil palm phenolics (OPP). The 2,2-diphenyl-1-picrylhydrazyl assay confirmed potent free radicalscavenging activity. To test for possible cardioprotective effects of OPP, we carried out in vitro LDL oxidation studies as well as ex vivoaortic ring and mesenteric vascular bed relaxation measurements. We found that OPP inhibited the Cu-mediated oxidation of humanLDL. OPP also promoted vascular relaxation in both isolated aortic rings and perfused mesenteric vascular beds pre-contracted with nor-adrenaline. To rule out developmental toxicity, we performed teratological studies on rats up to the third generation and did not find anycongenital anomalies. Thus, these initial studies suggest that OPP is safe and may have a protective role against free radical damage, LDLoxidation and its attendant negative effects, as well as vascular constriction in mitigating atherosclerosis. Oil palm vegetation liquor thusrepresents a new source of phenolic bioactives.Key words: Oil palm phenolics: Caffeoylshikimic acid: Antioxidant activity: TeratologyReactive oxygen species such as superoxide anions, H2O2and hydroxyl radicals may contribute to the genesis of CHD,diabetes, cancer and other degenerative diseases(1 – 5). Manyepidemiological studies have indicated that consumptionof fruit and vegetables decreases the risk of degenerativediseases(6,7), and that the beneficial effects, in part, can beascribed to the antioxidant activities of minor phytochemicalcomponents, including phenolic compounds(6 – 10).The oil palm (Elaeis guineensis) from the family Arecaceaeis a high oil-producing tropical plant that appears to have aneffective antioxidative component to counter the oxidativestress exerted by high temperature and intense sunlight.Indeed, the oil palm is a rich source of phytochemicals(11 – 13).While the technology for recovery of fat-soluble antioxidantssuch as tocopherols, tocotrienols and carotenoids frompalm oil is well established(11), it is only recently that thetechnology for harvesting water-soluble antioxidants from oilpalm has become available(12,14 – 17).During the palm oil milling process, water-soluble pheno-lics are discarded in the waste stream, amounting to 85 milliontons per year globally. A recovery procedure for oil palm phe-nolics (OPP) has been developed to isolate concentrations(14)suitable for biological applications, providing an opportunityto transform a bioburden into a range of potential applicationsfor health and wellness.Based on growing evidence that plant phenolics are ben-eficial to health, OPP was assessed for positive bioactivities.In the present study, the chemical constituents and compo-sition of specific OPP are described. In addition, in vitro anti-oxidant and LDL oxidation experiments, as well as ex vivoaortic ring and mesenteric vascular bed experiments, weredesigned to identify the potential bioactivities of OPP.* Corresponding author: R. Sambanthamurthi, fax þ 60 3 8926 1995, email [email protected]: DPPH, 2,2-diphenyl-1-picrylhydrazyl; OPP, oil palm phenolics.British Journal of Nutrition (2011), page 1 of 9 doi:10.1017/S0007114511002121q The Authors 2011British Journal of NutritionWe also carried out teratological studies to assess whetherOPP is safe for consumption.Materials and methodsHPLC, MS and NMR analyses of oil palm phenolicsOPP obtained according to the methods described bySambanthamurthi et al.(14)was subjected to separation byreversed-phase HPLC, and the individual peaks were charac-terised by MS and NMR spectroscopy. A freeze-dried OPPsample (10 mg) was dissolved in 1 ml of internal standardcitric acid solution (a-cyano-hydroxycinnamic acid (1 mg/ml)in 0·2 % (v/v) citric acid). The OPP sample was then extractedby adding 1 ml of ethyl acetate, shaking the solution well andthen allowing it to settle. The supernatant (200 ml) was evap-orated and reconstituted with 200 ml of 0·2 % (v/v) citricacid. This reconstituted solution was then injected into an ana-lytical HPLC system. Each compound in the sample was deter-mined using the peak ratio of the compound v. the internalstandard. The calibration curve was used to obtain the con-centration of each compound based on their peak ratios.Samples were analysed on a Hitachi system comprising alow-pressure mixing pump (model L7100; Hitachi, Richmond,CA, USA), an autosampler (model L7200; Hitachi), a photo-diode detector (L7450; Hitachi) and D-7000 HPLC systemsoftware for integration (Hitachi). Chromatographic separationwas achieved using a 250 £ 4·0 mm reversed-phase column(GL Exsil ODS 5 mm inner diameter) (SGE Inc., Austin, TX,USA). The mobile phase used was a binary gradient system,with phase A comprising 10 mM-sodium sulphate containing0·02 % (v/v) phosphoric acid (pH 2·75) and phase B compris-ing methanol–acetonitrile (70:30, v/v). Sample injectionvolume was 10 ml and a flow rate of 0·8 ml/min was used.The gradient elution with a total run time of 60 min was asfollows: started from 95 % (v/v) solvent A and 5 % (v/v) sol-vent B, increased to 35 % (v/v) solvent B over 45 min, thenincreased to 100 % (v/v) solvent B


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