Enhanced extraction of oleoresin from Piper nigrum by supercritical carbon dioxide using ethanol as a co‐solvent and its bioactivity profile

Pepper oleoresin was extracted using supercritical fluid extraction (SCF) from Piper nigrum berries. The conditions for maximum extraction of piperine were arrived at by calculating the solubility parameters of piperine and carbon dioxide using the Hildebrandt solubility equation. The effect of entrainers such as methanol, ethanol, and acetone was evaluated using theoretical modeling. The results predicted therefrom were validated experimentally. SCF at 250 bar/50 °C/60 min gave maximum relative extraction of 61.7 and 87.61% of oleoresin and piperine, while addition of 30% (vol/wt) ethanol as co‐solvent enhanced the yield to 95.9 and 184.7% under similar conditions of SCF extraction as compared to ethanol extraction for 6 hr. The oleoresins obtained showed comparable antioxidant, anti‐inflammatory, and antimicrobial activities with their respective controls. Hence SCF extraction of pepper using ethanol as a co‐solvent promises a rapid and enhanced recovery of oleoresin and its constituents with a good bioactivity profile.

Practical applications

Although SCF is a promising alternative for extraction of spice oleoresins, high cost of operation limits the use of this technology. In this work, process intensification of SCF extraction of black pepper lead to one step extraction and purification, along with high yield of oleoresin and the principle component piperine, eventually decreasing the cost of production. Thus, the highly concentrated SCF extracted oleoresins can have better replacement value for whole spices and better distribution in food and pharmaceutical sector.     

Antimicrobial and antioxidant activities of carvacrol microencapsulated in hydroxypropyl-beta-cyclodextrin

Carvacrol is a hydrophobic compound that exhibits high antimicrobial and antioxidant activities. Cyclodextrins are used to increase solubility and dissolution through water-insoluble compounds inclusion into their hydrophobic cavities. Hence, this study aimed to characterize carvacrol inclusion complexes with hydroxypropyl-beta-cyclodextrin (HPBCD) by establishing the physico-chemical properties and evaluate their antimicrobial and antioxidant properties. Inclusion complexes were prepared by kneading (KN) and freeze-drying (FD) methods. Entrapment efficiency was 78.09 ± 1.24% for KN, and 83.74 ± 1.15% for FD. Polydispersity index was greater than 0.2 for both methods. Particle size for KN and FD were 0.360 ± 0.003 and 0.377 ± 0.007 μm, respectively. Carvacrol–HPBCD antimicrobial activity was higher (P < 0.05) than for free carvacrol for both bacteria, Escherichia coli K12 and Salmonella enterica serovar Typhimurium LT2, indicating that HPBCD increased water solubility and consequently increased contact between carvacrol and bacteria in medium. Antioxidant activity was lower (P < 0.05) for inclusion complexes indicating HPBCD makes carvacrol less available to react with free radical. The stability study indicated that light did not affect (P > 0.05) degradation, indicating that the microparticles were stable throughout storage. Therefore, carvacrol–HPBCD complexes may have important applications in the food industry as stable antimicrobial systems.     

International Regulations of Propolis Quality: Required Assays do not Necessarily Reflect their Polyphenolic‐Related In Vitro Activities

Propolis has been proposed as a polyphenolic‐rich natural product potentially able to be used for human consumption or even for medicinal proposes. To guarantee a minimum phenolic and flavonoid content and as consequence of their related‐biological activities, international requirements of propolis quality are commonly applied. In this work we assessed phenolic and flavonoid contents of propolis; the antioxidant capacity (toward peroxyl radicals and hypochlorous acid); the ability to generate nitric oxide (NO); and, finally the antimicrobial activity of 6 propolis samples from the VI region of Chile. Our results show that the total phenolic and flavonoid content of propolis samples are not always in agreement with their polyphenolic‐associated in vitro activities. For example, P03 and P06 samples showed the lowest (25 ± 4 GAE/g propolis) and the highest (105 ± 3 GAE/g propolis) total phenolic content, respectively. This was in agreement with flavonoid content and their Oxygen Radical Absorbance Capacity (ORAC) activity. However, this dependence was not observed toward HOCl, NO release and antimicrobial activity. Based on our results, we consider that, in order to guarantee the antioxidant or antimicrobial in vitro effects, the international regulations of propolis quality should contemplate the convenience of incorporating other simple analytical test such as ORAC or antimicrobial tests.     

Jabuticaba (Myrciaria cauliflora) Seeds: Chemical Characterization and Extraction of Antioxidant and Antimicrobial Compounds

This study was aimed to assess the effect of time and temperature on the extraction of antioxidant compounds from jabuticaba seeds (Myrciaria cauliflora cv. Sabará), to optimize the solvent proportion (water, ethyl alcohol, and propanone), and to characterize the extract according to the chemical composition, antioxidant, and antimicrobial properties. Proximal composition, total phenolic content (TPC), antioxidant, and antimicrobial activities were analyzed. The optimized solvent ratio of 60% water and 40% propanone provided a mean TPC of 8.65 g GAE/100 g seeds and the antioxidant activity toward 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) was 82.79% ± 0.50%. Time and temperature parameters did not influence the yield of TPC. The gross seed extract was partially purified and both exhibited a high antioxidant activity and antimicrobial potential toward Gram‐positive and Gram‐negative bacteria. The purified jabuticaba seed lyophilized extract contained a higher (P < 0.05) TPC, o‐diphenols, flavonols, and antioxidant activity measured by the DPPH assay and total reducing capacity as compared to the gross lyophilized extract. Electrospray ionization coupled with tandem mass spectrometry (ESI‐MS/MS) data showed the presence of ellagitannins and ellagic acid in the extracts, which are probably the responsible for the antimicrobial and antioxidant activities.     

ANTIOXIDATIVE AND ANTIBACTERIAL POTENTIALS OF ESSENTIAL OILS AND EXTRACTS ISOLATED FROM VARIOUS SPICE MATERIALS†

The antioxidant and antibacterial potentials of essential oils and acetone extracts of black pepper, cumin, black cumin and mace were carried out by different techniques. The antioxidative capacity of the essential oils and acetone extracts were evaluated against mustard oil by measuring peroxide and thibarbituric acid values at fixed intervals. In addition, their antioxidant potential was evaluated by 2, 2′‐diphenyl‐1‐picrylhydracyl radical and conjugated diene assays. Their reducing power was determined with standards, which proved the strong antioxidant capacity of essential oils and extracts. The antioxidant activity of essential oils and extracts exerted by all the antioxidant assays can be compared with synthetic antioxidants such as butylated hydroxyanisole and butylated hydroxytoluene. The antibacterial activity was studied by disk diffusion and poison food methods. Black cumin essential oil showed complete zone of inhibition (P < 0.05) against tested bacterial strains of Staphylococcus aureus, Bacillus cereus and Bacillus subtilis at 2 and 6 µL level by disk diffusion method. Black cumin and black pepper extracts showed complete reduction of colonies against tested bacterial strains of S. aureus, B. cereus and B. subtilisat 5 and 10 µL level by poison food method. Poison food method exhibited good results for the tested essential oils and extracts. Essential oils of black pepper, cumin, black cumin and mace may be used to stabilize mustard oil after screening.     

Physical,antimicrobial and antioxidant properties of starch‐based film containing ethanolic propolis extract

Propolis is a natural product that meets the requirements as functional additive for food packaging due to its antioxidant and antimicrobial activities. In this work, ethanolic propolis extract (EPE) was incorporated in cassava starch films, and characterisations with respect to their microstructure, mechanical properties, water vapour permeability (WVP), moisture sorption kinetics as well as antimicrobial and antioxidant capacities were performed. The results showed that tensile strength was not affected (P > 0.05) by the presence of EPE but Young's modulus decreased about 50% when compared to control films possibly because of EPE plasticiser effect. When 1% EPE was used, changes in moisture sorption properties were detected by a slightly hydrophobic character at films WVP. When extracted from the films, propolis retained its antioxidant activity. The films exhibited antimicrobial activity against Staphylococcus aureus and Escherichia coli even at low EPE concentrations (0.5%) mainly due to its phenolic compounds.     

The Phenolic Compounds,Metabolites, and Antioxidant Activity of Propolis Extracted by Ultrasound‐Assisted Method

The influences of ultrasound‐assisted, pharmacopeia, and supercritical fluid extraction methods on bioactive compounds and biological activities of propolis were evaluated. Results showed that propolis extracted by ultrasound‐assisted method contained more phenolic compounds, and showed the highest total phenolic content (245.84 ± 6.41 mg GAE/g DW), total flavonoids content (198.82 ± 5.74 mg RE/g DW), and stronger in vitro antioxidant activity (DPPH·: 1.03 ± 0.04 mmol Trolox/g DW, ABTS+·: 2.19 ± 0.05 mmol Trolox/g DW, and FRAP: 1.48 ± 0.12 mmol FeSO₄/g DW) than those of pharmacopoeia and supercritical fluid methods. A total of 36 phenolic compounds were identified in propolis. Among them, quercetin, quercetin‐3‐methyl‐ether, kaempferol, isorhamnetin, luteolin‐methyl‐ether, and quercetin‐7‐methyl‐ether could only be found in ultrasound‐assisted and pharmacopoeia methods. Moreover, the phenolic compounds had the similar metabolic pathways in rats and were mainly metabolized by sulfation and glucuronidation pathways. Additionally, ultrasonic‐treated propolis have good in vivo antioxidant activity and could repair D‐galactose‐induced oxidative damage in rats. Therefore, ultrasound‐assisted method could replace pharmacopeia method to be considered as bioactive compounds extraction from propolis, taking into consideration of yield, short extraction time, and high antioxidant activity.     

Piperine—The Bioactive Compound of Black Pepper: From Isolation to Medicinal Formulations

Piperine is the major bio‐active component of pepper, which imparts pungency and biting taste to it. This naturally occurring alkaloid has numerous demonstrated health effects and beneficial therapeutic properties; nevertheless, its biological applications are limited due to its poor solubility in aqueous environments. This emphasizes an implementation of advanced extraction approaches which could enhance the extraction yield of piperine from pepper and also the development of new formulations containing piperine to improve its in vivo bioavailability. This paper presents a review on the therapeutic and medicinal effects of piperine, its isolation from pepper fruit and the development of new formulations for its medicinal (pharmaceutical) applications. A thorough review on conventional and advanced separation techniques for the extraction of piperine from pepper is presented and an outline of the most significant conditions to improve the extraction yield is provided and discussed. Different methods used to measure and quantify the isolated piperine are also reviewed. An overview of biotechnological advancements for nanoparticle formulations of piperine or its incorporation in lipid formulations, which could enhance its bioavailability, is also presented.     

p‐Coumaric acid and its conjugates: dietary sources,pharmacokinetic properties and biological activities

p‐Coumaric acid (4‐hydroxycinnamic acid) is a phenolic acid that has low toxicity in mice (LD₅₀ = 2850 mg kg^?1 body weight), serves as a precursor of other phenolic compounds, and exists either in free or conjugated form in plants. Conjugates of p‐coumaric acid have been extensively studied in recent years due to their bioactivities. In this review, the occurrence, bioavailability and bioaccessibility of p‐coumaric acid and its conjugates with mono‐, oligo‐ and polysaccharides, alkyl alcohols, organic acids, amine and lignin are discussed. Their biological activities, including antioxidant, anti‐cancer, antimicrobial, antivirus, anti‐inflammatory, antiplatelet aggregation, anxiolytic, antipyretic, analgesic, and anti‐arthritis activities, and their mitigatory effects against diabetes, obesity, hyperlipaemia and gout are compared. Cumulative evidence from multiple studies indicates that conjugation of p‐coumaric acid greatly strengthens its biological activities; however, the high biological activity but low absorption of its conjugates remains a puzzle. © 2015 Society of Chemical Industry     

Correlation between Antimicrobial,Antioxidant Activity,and Polyphenols of Alkalized/Nonalkalized Cocoa Powders

Many factors can influence antioxidative and antimicrobial characteristics of plant materials. The quality of cocoa as functional food ingredient is influenced through its processing. The main aim of this study was to test if there is difference in polyphenol content, antioxidant capacity, and antimicrobial activity between nonalkalized and alkalized cocoa powders. To estimate polyphenol and flavonoid content in cocoa samples the spectrophotometric microassays were used. Flavan‐3ols were determined with reversed‐phase high‐performance liquid chromatography (RP‐HPLC). Antimicrobial activity against 3 Gram positive bacteria, 4 Gram negative bacteria and 1 strain of yeast was determined using broth microdilution method. Total polyphenol content was 1.8 times lower in alkalized cocoa samples than in natural ones. Epicatechin/catechin ratio was changed due to the process of alkalization in favor of catechin (2.21 in natural and 1.45 in alkalized cocoa powders). Combined results of 3 antioxidative tests (DPPH, FRAP, ABTS) were used for calculation of RACI (Relative Antioxidant Capacity Index) and GAS (Global Antioxidant Score) values that were consistently higher in natural than in alkalized cocoa extracts. Obtained results have shown significant correlations between these values and phenolic content (0.929 ≤ r ≤ 0.957, P < 0.01). Antimicrobial activity varied from 5.0 to 25.0 mg/ml (MICs), while Candida albicans was the most sensitive tested microorganism. Cocoa powders subjected to alkalization had significantly reduced content of total and specific phenolic compounds and reduced antioxidant capacity (P < 0.05), but their antimicrobial activity was equal for Gram‐positive bacteria or even significantly enhanced for Gram‐negative bacteria.     

Effect of natural biostimulants on yield and nutritional quality: an example of sweet yellow pepper (Capsicum annuum L.) plants

BACKGROUND: Modifications in growing techniques can affect the yield and nutritional quality of various cultivated plant species. Owing to its high nutritional value, pepper (Capsicum annuum L.) was used in this study as a model plant to investigate the effect of natural biostimulants on yield and fruit quality parameters under conditions of reduced fertilisation. RESULTS: A positive influence of biostimulant treatment on yield parameters was observed. The overall increase in the pigment content of leaves after biostimulant application agreed well with the higher total and commercial yields of treated pepper cultivars compared with their controls. The results showed that natural biostimulants had a positive effect on the vitamin C and total phenolic contents in pepper fruits during the hot summer season. The 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) and 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonate) (ABTS) antioxidant activities were also significantly higher (P < 0.05) in treated plants and correlated strongly with all measured quality parameters except total phenolic content. CONCLUSION: Generally, biostimulants improved the antioxidant activity, vitamin C and phenolic contents in fruits as well as the pigment content in leaves of treated compared with non‐treated pepper plants grown hydroponically. Thus the application of biostimulants could be considered as a good production strategy for obtaining high yields of nutritionally valuable vegetables with lower impact on the environment. Copyright © 2011 Society of Chemical Industry     

Microencapsulation of black pepper oleoresin

Despite of solvent extracted spice oleoresin having many advantages over ground spices, its sensitivity to light, heat and oxygen is a disadvantage. One approach to overcome this is microencapsulation. The present work reports on microencapsulation of black pepper oleoresin by spray-drying, using gum arabic and modified starch as wall materials. The microcapsules were evaluated for the content and stability of volatiles, non-volatiles, total piperine and entrapped piperine for six weeks. Gum arabic offered greater protection to the pepper oleoresin than modified starch, as seen from the t_1/2, time required for a constituent to be reduced to 50% of its initial value.     

Drying kinetics,physico‐chemical properties,antioxidant activity and phenolic composition of foam‐mat dried germinated rice bean (Vigna umbellata) hydrolysate

This research was aimed to study physico‐chemical properties and antioxidant activities of foam‐mat dried germinated rice bean (Vigna umbellata) hydrolysate. Germination led to an increase in released phenolic content and antioxidant activity (DPPH radical scavenging activity and FRAP) of rice bean hydrolysate. The hydrolysate obtained from germinated rice bean (GRB) and non‐germinated rice beans (NGRBs) was foam‐mat dried at 60 and 70 °C. Semi‐theoretical and empirical model could suitably describe the drying characteristic of foamed bean hydrolysate. Total phenolic content and antioxidant activities of foam‐mat dried samples decreased with increasing air‐drying temperature (P ≤ 0.05). Gallic acid, catechol and epicatechin were major phenolic compounds in foam‐mat dried samples prepared from both GRB and NGRB. The higher phenolic content and antioxidant activities were found in foam‐mat dried hydrolysate of GRB. Electron spin resonance spectrometry revealed that foam‐mat dried rice bean hydrolysate showed a strong ability to scavenge free radicals, especially carbon‐centred radicals.     

Antimicrobial Activities of Phenolic Extracts Derived from Seed Coats of Selected Soybean Varieties

Soybean hulls or seed coats consist of complex carbohydrates, proteins, lipids, and polyphenols such as anthocyanidins, proanthocyanidins, and isoflavones. The polyphenolics in the seed coats give them various colors such as black, brown, green, yellow, or even a mottled appearance. In this study, the antimicrobial effects of phenolic extracts from the seed coats of different colored soybeans (yellow, dark brown, brown, and black) were evaluated against foodborne pathogens such as Salmonella Typhimurium, Escherichia coli O157:H7, and Campylobacter jejuni in broth‐cultures as well as on chicken skin. The highest total phenolic content was observed for the phenolic extract from soybean variety (R07‐1927) with black colored seed coat (74.1 ± 2.1 mg chlorogenic acid equivalent [CAE]/g extract) and was significantly different (P <0.0001) from the extract of the conventional soybean variety (R08‐4004) with yellow colored seed coat (7.4 ± 1.2 mg CAE/g extract). The extract from black colored soybean produced reductions of 2.10 ± 0.08 to 2.20 ± 0.08‐log CFU/mL for both E. coli O157:H7 and C. jejuni after 3 d when incubated in broth‐culture having 4‐log CFU/mL of bacteria, whereas a 6 d incubation was found to reduce S. Typhimurium and E. coli O157:H7 at 2.03 ± 0.05 and 3.3 ± 0.08‐log CFU/mL, respectively. The extract also reduced S. Typhimurium and E. coli O157:H7 attached to chicken skin by 1.39 ± 0.03 and 1.24 ± 0.06‐log CFU/g, respectively, upon incubation for 6 d. Soybean seed coat extracts may have a potency as antimicrobial agents to reduce foodborne bacteria contaminating poultry products.     

Antioxidant and antimicrobial activities of Polygonum cognatum Meissn extracts

The antioxidant activities, reducing powers, 2,2‐diphenyl‐l‐picrylhydrazyl (DPPH) radical‐scavenging activities, total phenolic compound contents and antimicrobial activities of ether, ethanol and hot water extracts of Polygonum cognatum Meissn were studied in vitro. The highest antioxidant activity was found in the water extract. However, there were no statistically significant differences among 15 µg ml^?1 extract‐containing samples in linoleic acid emulsion (0.02 M , pH 7.0) during 120 h of incubation (P > 0.05). The reducing power of the water extract was the highest, but its reducing power was markedly lower than that of ascorbic acid. The highest DPPH radical‐scavenging activity was found in the water extract, with 50% DPPH radical scavenging at a concentration of 100 µg ml^?1 dried water extract, while at the same concentration of dried ethanol extract the value was 12%. Surprisingly, no DPPH radical‐scavenging activity was observed in the ether extract. The concentrations of phenolic compounds found were 0.48, 0.50 and 0.01 µg ml^?1 gallic acid equivalent in 10 µg ml^?1 water, ethanol and ether extracts respectively. The ether and ethanol extracts showed antimicrobial activity against Staphylococcus aureus and Bacillus subtilis. The water extract did not show antimicrobial activity against the studied micro‐organisms. © 2002 Society of Chemical Industry     

Physicochemical properties,antioxidant activity and inhibition of α‐glucosidase of a novel fermented pepper (Capsiccum annuum L.) leaves‐based vinegar

The aim of this study was to utilise by‐product pepper leaves for vinegar production due to its nutritional value. Changes in physicochemical and functional properties in four main process from raw material (leaves) to lactic acid (juice), alcohol (wine) and acetic acid (vinegar) fermentations were evaluated and compared. Their nutritional compositions were significantly different and particularly the wine and vinegar had higher total phenol content and their expanded diversity than the juice and then leaves (P < 0.05). Difference in values for phenolic compound was highly correlated with their α‐glucosidase inhibitory (AGI) and antioxidant capacities in DPPH and superoxide radical scavenging and reducing power assays (r = 0.919–0.981); vinegar ≥ wine > juice > leaves. Especially, AGI activity was greatly enhanced depending on fermentation steps beyond expectation. Thus, these data confirm that pepper leaves is a good functional source and the vinegar fermentation improved the functionality in stages.     

Chemical,antioxidant and antibacterial study of Brazilian fruits

In this study, the antioxidant capability, total phenolic content and antimicrobial activity of ethanolic extracts of seven fruits from the Brazilian Atlantic Forest were evaluated. The conditions for the extraction of crude phenolics from the fruits were determined using an experimental factorial design. Total phenolic content, 1,1‐diphenyl‐2‐picrylhydrazyl radical (DPPH^?) scavenging activity and β‐carotene‐linoleic acid couple oxidation assays were used to evaluate the antioxidant properties of the extracts. In addition, antimicrobial activity was screened using two Gram‐negative bacteria (Escherichia coli and Klebsiella pneumoniae) and one Gram‐positive bacterium (Staphylococcus aureus). All native fruits assayed in this study have high potential as natural antioxidant sources. Among the seven fruits evaluated, Jabuticaba and Uvaia had the highest antioxidant activity in the DPPH^? and of β‐carotene‐linoleic acid coupled oxidation assays. In the biological assay, K. pneumoniae was the most sensitive microorganism to the fruit extracts, and the Jabuticaba extract had a slight inhibitory effect against this Gram‐positive bacterium.     

Evaluation of the Phenolic Content,Antiradical, Antioxidant,and Antimicrobial Activity of Different Floral Sources of Honey

Thirty-five honeys were evaluated for total phenolic content by Folin-Ciocalteu method, for potential antioxidant activity using phosphomolibdenum assay and by the 1,1-diphenyl-2-picrylhydrazyl method for antiradical activity. The antimicrobial activity was studied by the agar diffusion method, using 12 bacteria and 2 yeasts. The means of the total phenolic contents of chestnut, honeydew, multifloral, thyme, and astragalus were 47 ± 18, 24.2 ± 0.6, 14 ± 11, 11 ± 6, and 9 ± 7 mg/100 g honey as gallic acid equivalent, respectively. The lowest antioxidant activity was observed in honeydew 70 ± 5 mg ascorbic acid equivalent/g honey while the highest content was observed in astragalus honey 86 ± 16 mg ascorbic acid equivalent/g honey. Correlation between the phenolic content and antioxidant activity were found to be statistically significant. Chestnut honeys (n = 5) exhibited maximum free radical scavenging activity with an average 68 ± 9%. The honey samples showed the highest antimicrobial activity against some microorganisms, especially Escherichia coli O157:H7, Salmonella typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Proteus mirabilis. On the other side, Aeromonas hydrophila, Bacillus subtilis, and E. coli were the most resistant microorganisms. The results revealed that the Turkish honeys studied proved to be a good source of antioxidant and antimicrobial agents that might serve to protect human health.     

Supercritical CO₂extraction of oleoresin from marigold (Tagetes erecta L.) flowers and determination of its antioxidant components with online HPLC-ABTS(·+) assay

BACKGROUND: Marigold is a traditional medicine herb which shows good pharmacological activity in many aspects. It is very important to obtain and investigate the specific bioactive compounds from marigold. The objective of the study was to extract the oleoresin from marigold with supercritical CO₂ (SC‐CO₂) at different pressures and temperatures, detect the fatty acid composition by gas chromatography–mass spectrometry and investigate the antioxidative components in the extracts by combined online high‐performance liquid chromatography‐2,2‐azinobis‐(3‐ethylbenzothiazolin‐6‐sulfonic acid (HPLC‐ABTS^?+) post‐column assay and HPLC‐tandem mass spectrometry. RESULTS: For the pressure range (20–40 MPa) and temperature range (30–70 °C), 30 MPa/70 °C gave the highest yield of oleoresin (58.9 g kg^?1). The dominant fatty acids of marigold flower oleoresin were linoleic acid (>26.41%), palmitic acid (>24.22%) and oleinic acid (>20.12%). Significant effects of the extraction pressure and temperature on the antioxidant activity were observed (P < 0.05). Lutein esters, α‐tocopherol, β‐tocopherol, γ‐tocopherol and δ‐tocopherol were the dominant antioxidant compounds in the extracts. CONCLUSION: The study has shown that the yield and total antioxidant activity of the marigold extracts were affected by the pressure and temperature of SC‐CO₂, and that online HPLC technique could be used as an efficient and rapid method for separation and identification of bioactive compounds from a complex mixture. Copyright © 2011 Society of Chemical Industry