Enzymatic conjugation with fatty acids including omega-3 polyunsaturated fatty acids (ω-3 PUFAs) derived from fish oil to three citrus fruit-derived flavonoids: grapefruit extract, naringin, and neohesperidin dihydrochalcone were investigated. The conversions were achieved over 85% under the catalysis of lipase Novozyme 435 in acetone at 45°C at semi-preparative scale. The conjugates were purified via solvent partition and silica gel chromatography and achieved 90-98% in purity. The NMR analysis of the conjugates confirmed that the fatty acid carbon chain was linked onto the primary -OH group on the glucose moiety of the flavonoids. The purified flavonoid conjugates alongside their original flavonoids were analyzed for antioxidant activities via 2,2-diphenyl-1-picrylhydrazyl scavenging assay, and anti-peroxidation test via peroxide values measured during a 1-week fish oil storage trial. Vascular endothelial growth factor (VEGF) assay was conducted with 1, 10, and 100 μM of naringin and grapefruits and their conjugates, respectively, and total VEGF levels were measured at 24 and 48 h, respectively, using ELISA and dot blot analysis. The results from these functionality experiments demonstrated that flavonoid FA conjugates have at least comparable (if not higher) antioxidant activity, anti-peroxidation activity, and anti-angiogenic activity.

Passion fruit peel extract (PFP) was used in a randomized, double-blind, placebo-controlled clinical trial to investigate its potential effect on knee osteoarthritis (OA) symptoms. Thirty-three OA patients were enrolled. They received either placebo or PFP pills (150 mg, daily) by evaluating monthly with Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index. The total and subscale WOMAC scores significantly improved in the PFP group. At 60 days, reductions of 18.6, 18, 19.6, and 19.2 percent in self-reported pain, stiffness, physical function, and composite WOMAC scores, respectively, were reported in the PFP group, whereas in the placebo group, WOMAC scores increased in every category. The results of this study indicated the efficacy of PFP in alleviating osteoarthritis symptoms.

Knee osteoarthritis (OA) is a common degenerative joint disorder and a major cause of pain and disability. The hypothesis tested in this study was that the passion fruit peel extract (PFP), a flavonoid-rich dietary supplement, would reduce symptoms due to knee OA. Thirty-three OA patients were enrolled in a randomized, double-blind, placebo-controlled trial with parallel-group design. Patients received either placebo or PFP pills (150 mg, daily) in a double-blinded fashion for 2 months. The OA clinical symptoms were evaluated monthly with Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index. In the PFP group, there was a significant improvement in total WOMAC score and WOMAC subscale score of physical function after 30 days and pain after 60 days. At 60 days, reductions of 18.6%, 18%, 19.6%, and 19.2% in pain, stiffness, physical function, and composite WOMAC score, respectively, were self-reported in the PFP group. Whereas, in the placebo group, the self-reported WOMAC scores increased in every category. The results of this study show that PFP substantially alleviated osteoarthritis symptoms. This beneficial effect of PFP may be due to its antioxidant and antiinflammatory properties.

With an increasing number of patients suffering from asthma and the frequent inability of conventional life style modification and therapy to effectively control the problem, nutritional and dietary therapies are being sought. This study was undertaken to investigate the efficacy of the purple passion fruit peel (PFP) extract, a novel mixture of bioflavonoids, on asthma symptoms. Humans with asthma were studied in a four-week randomized, placebo-controlled, double-blind trial with oral administration of PFP extract (150 mg/day) or placebo pills. The effects of PFP extract were evaluated by assessing the clinical symptoms of asthma and spirometry tests. Most clinical symptoms of asthma of the PFP extract-treated group were moderated significantly compared to those taking placebo. The prevalence of wheeze, cough, as well as shortness of breath was reduced significantly in group treated with PFP extract (P < .05), while the placebo caused no significant improvement. No significant difference were observed in the forced expiratory volume at one second and forced vital capacity of those supplemented were with PFP extract. No adverse effect was reported by any of study participants. The PFP extract may be safely offered to asthmatics as an alternative treatment option to reduce clinical symptoms.

Asthma, affecting as many as 400 million individuals worldwide, is one of the most prevalent chronic health condition in the United States. With an increasing number of patients with asthma and the frequent inability of conventional lifestyle modification and therapy to effectively control the problem, nutritional and dietary therapies are being sought. This study was undertaken to investigate the efficacy of the purple passion fruit peel (PFP) extract, a novel mixture of bioflavonoids, on asthma symptoms. Patients with asthma were studied in a 4-week randomized, placebo-controlled, double-blind trial with oral administration of PFP extract (150 mg/d) or placebo pills. The effects of PFP extract were evaluated by assessing the clinical symptoms of asthma and spirometry tests. Most clinical symptoms of asthma of the PFP extract-treated group were moderated significantly compared to the baseline. The prevalence of wheeze, cough, as well as shortness of breath was reduced significantly in group treated with PFP extract (P < .05), whereas the placebo caused no significant improvement. Purple passion fruit peel extract supplementation resulted in a marked increase in forced vital capacity (P < .05) as placebo showed no effect. However, no significant improvement was observed in the forced expiratory volume at 1 second of those supplemented with PFP extract. No adverse effect was reported by any of study participants. The PFP extract may be safely offered to asthmatic subjects as an alternative treatment option to reduce clinical symptoms.

Hypertension is one of the most important modifiable risk factors for cardiovascular and cerebrovascular diseases. We investigated the potential antihypertensive effect of the purple passion fruit peel (PFP) extract, a mixture of bioflavonoids, phenolic acids, and anthocyanins, in spontaneously hypertensive rats and human. A high-performance liquid chromatography analysis was performed to identify the active ingredients of the PFP extract. In a rat liver toxicity assay, no hepatotoxicity was observed after 9 hours incubation in the presence of PFP extract (20 μg/mL). The PFP extract also revealed hepatoprotection against chloroform (1 mmol/L)-induced liver injury. In the experimental model, 24 spontaneously hypertensive rats were divided into 3 treatment groups for a period of 8weeks: control group and groups fed diet supplemented with either 10 or 50 mg/kg PFP extract. The diet supplemented with PFP extract at 50 mg/kg significantly lowered systolic blood pressure by 12.3mm Hg (P < .01) and markedly decreased serum nitric oxide level by 65% (P < .05) compared with the control group. The effect of the treatment on immune parameters was also evaluated, which showed no statistical changes. Studies were then extended to hypertensive human subjects who were administered PFP extract (400 mg/d) or placebo pills within a 4-week randomized, placebo-controlled, double-blind trial. The effects of the PFP extract were evaluated by blood pressure measurement. The systolic and diastolic blood pressure of the PFP extract–treated group decreased significantly by 30.9 ± 6.3 and 24.6 ± 3.3 mm Hg, respectively, compared with the placebo group (P < .001). No adverse effect was reported by the patients. The results suggest that the antihypertensive effect of the PFP extract may, in part, be mediated through nitric oxide modulation. It is suggested that the PFP extract may be offered as a safe alternative treatment to hypertensive patients.

Effects of particle size, temperature, contact time, solvent-to-sage ratio and the ethanol–water ratio on the extraction of the active compounds rosmarinic acid, carnosic compounds and essential oil from dried sage (Salvia officinalis) were studied. Optimal extraction conditions giving highest yield of all three active compounds were particle diameter 1mm, extraction temperature 40°C, solvent-to-sage ratio of 6:1 and 55–75wt% ethanol for up to 3h. This gave an extract equivalent to 14.9% of dry sage, containing 6.9% rosmarinic acid (55% recovery), 10.6% carnosic compounds (75% recovery) and 7.3% essential oil (42% recovery). Scale up of the process by a factor of 100 demonstrated that the optimised laboratory scale process can be carried out without any loss of efficiency at an industrial scale.

An acetone:water (7:3) extract obtained from the leaves of Rumex obtusifolius was fractionated into procyanidin oligomer and polymer fractions using a linear gradient and a simple step method on Sephadex LH-20. The chemical characteristics of the procyanidin fractions were studied by C-13-NMR spectroscopy, acid-catalysed degradation in the presence of benzyl mercaptan, matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) MS and electrospray ionisation (ESI) MS. The C-13-NMR showed that the polymer fraction consisted predominantly of procyanidin polymers, some with galloyl groups attached. The thiolysis reaction products indicated a mean degree of polymerisation (DP) of 4.3 for the step method, and a range of 2.3-8.2 mean DP for the gradient fractionation., with epicatechin as the most abundant flavan-3-ol extension unit, while the terminal units consisted of equal proportions of catechin, epicatechin and epicatechin gallate. Singly charged ions observed in MALDI-TOF/MS showed a range of oligomeric procyanidins and their polygalloyl derivatives. These species (in the range DP 2-7) were also observed by ESI/MS but the spectra were more complex due to overlapping multiply charged ions. Isolation of oligomers from the Sephadex LH-20 fraction by chromatography on polyamide and C-18 yielded B1, B2, B3 and B7 dimers, an A-type trimer and a B2 3,3'-O-digallate. Copyright (c) 2007 John Wiley & Sons, Ltd.

The negative-ion fast-atom-bombardment (FAB) mass spectra of procyanidin oligomers are characterized by abundant [M-H]-ions and fragmentation that results primarily from retro-Diels-Alder fission and cleavage of the interflavanoid bond. B/E-linked scanning mass spectrometry established that sequence ions result from unimolecular gas-phase decompositions. A quinone-methide mechanism of interflavanoid bond cleavage is proposed to account for observed ions of different molecular weight originating from isomeric upper and lower flavan units. These ions can be used to establish the sequence of monomer units and also to distinguish a linear from a branched trimer.

With an increasing number of patients suffering from hypertension and the frequent inability of conventional diet modification and therapy to effectively control the problem, alternative therapies are being sought after. These studies were undertaken to investigate the efficacy of the purple passion fruit peel (PFP) extract—a mixture of bioflavonoids, phenolic acids, and anthocyanins—in animals and patients with mild to severe hypertension. Spontaneously hypertensive rats were divided into 3 groups: basic diet, basic diet supplemented with the PFP extract at 10 mg/kg and 50 mg/kg; and serum nitric oxide and systolic blood pressure were analyzed. Studies were then extended to hypertensive subjects within a four-week randomized, double-blind, placebo-controlled trial through the administration of oral PFP extract (400 mg/day) or placebo pills. In spontaneously hypertensive rats, consumption of the PFP extract at 50 mg/kg lowered systolic blood pressure by 12.3 mm Hg on average (p<0.01) and markedly decreased the nitric oxide production by 65% on average (p<0.05). In hypertensive humans, the systolic and diastolic blood pressure of those supplemented with the PFP extract decreased significantly (p<0.001). Hypertensive patients given the PFP extract reported experiencing no significant adverse effects from it. The purple passion fruit peel extract may be safely offered to hypertensive patients as an alternative treatment option. This study was supported in part by Kemin Foods.

Few EPR spectra of aroxyl radicals from model gallate esters and gallotannins and none for the more complex ellagitannins have previously been recorded. Hybrid density-functional calculations were carried out to determine isotropic coupling constants for the gallate ester radicals and gallotannins, confirming structures of both propylgallate mesomers and the hamamelitannin radical. This approach has been applied to the identification of the radicals observed after alkaline autoxidation of the ellagitannins amariin, corilagin and geraniin with either hexahydroxydiphenic acid and/or the dehydrohexahydroxydiphenic acid moieties. In case of amariin, the EPR spectrum reflects only the C(1), galloyl radical, whereas in case of corilagin and geraniin, thin most reasonable interpretation is that of a composite spectrum of both a C(1),galloyl radical and a hexahydroxydiphenic acid-derived semiquinone radical.

Some polyphenol-containing extracts (Pelargonium sidoides, Phyllanthus amarus) and representatives of simple phenols (shikimic acid 3- and 5-O-gallate), flavan-3-ols (epigallocatechin 3-gallate), proanthocyanidins (a hexamer) and hydrolysable tannins (corilagin, casuariin, geraniin) were studied for gene expressions (iNOS, IL-1, IL-10, IL-12, IL-18, TNF-alpha, IFN-alpha/gamma) by RT-PCR. All extracts and compounds were capable of enhancing the iNOS and cytokine mRNA levels in parasitised cells when compared with those in non-infected conditions.

Caffeic acid, rosmarinic acid and oligomers of caffeic acid with multiple catechol groups are all constituents of Salvia officinalis. Their antioxidant potential was investigated with regard to their radical scavenging activity and the stability and structure of the intermediate radicals. Pulse-radiolytic studies revealed very high rate constants with hydroxyl radicals. Evidence from kinetic modeling calculations suggested an unusual complex behavior due to the presence of both O4- and O3-semiquinones and formation and decay of a hydroxyl radical adduct at the vinyl side chain. The radical structures observed by EPR spectroscopy after autoxidation in slightly alkaline solutions were only partially identified due to their instability and generally represented dissociated O4-semiquinones. Hybrid density-functional calculations of the potential radical structures showed distinct differences between the resonance stabilization of the O4- and O3-semiquinones of caffeic and dihydrocaffeic acids, reflected also in the considerably faster decay of the O3-semiquinone observed by pulse radiolysis. No evidence was found for dimerization reactions via Cbeta radicals typical for lignin biosynthesis.

The chemical characteristics of the purified procyanidin polymers of the flowers of the forage legume red clover (Trifolium pratense L.) were studied by (13)C NMR, acid-catalyzed degradation with benzyl mercaptan, and electrospray ionization mass spectrometry (ESI-MS). The (13)C NMR showed that the fraction consisted of predominantly procyanidin polymers. The thiolysis reaction products indicated a mean degree of polymerization (mDP) of 9.3 with epicatechin (81%) as the abundant flavan-3-ol extension unit and the terminating units dominated by catechin (95%). ESI-MS showed a range of oligomeric procyanidin ions (DP of 2-11). The white clover floral prodelphinidins consist of terminal units with nearly equal proportions of epigallocatechin (52%) and gallocatechin (48%) and extender units showing epigallocatechin (56%) and gallocatechin (39%). The dramatic difference in the stereochemistry of the terminal and extender units observed for the red clover floral procyanidins contrasts with the mixture of cis and trans stereochemistry observed for white clover floral prodelphinidins.

Seven representative compounds isolated from Salvia officinalis, among them caffeic acid, the dimer rosmarinic acid and oligomers of caffeic acid, were investigated with regard to their antioxidant potential both expressed by the radical scavenging activity and the stability and structure of the intermediate radicals. Pulse-radiolytic investigation revealed very high rate constants with both hydroxyl and azide radicals. Evidence from kinetic modelling calculations suggested unusual complex behavior due to the presence of both O(4)- and O(3)-semiquinones and - in two cases - formation and decay of a hydroxyl radical adduct at the vinyl side chain. EPR spectroscopy studies, which included dihydrocaffeic acid as a model for the saturated side chains of the oligomers, confirmed that the radical structures after oxidation in slightly alkaline solutions are representing dissociated O(4)-semiquinones. While according to calculations based on hybrid density-functional theory the other radical structures are valid intermediates, they cannot be observed except by pulse radiolysis due to their fast decay.