Maritime Halophyte Species from Southern Portugal as Sources of Bioactive Molecules

Extracts of five halophytes from southern Portugal (Arthrocnemum macrostachyum, Mesembryanthemum edule, Juncus acutus, Plantago coronopus and Halimione portulacoides), were studied for antioxidant, anti-inflammatory and in vitro antitumor properties. The most active extracts towards the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical were the methanol extracts of M. edule (IC₅₀ = 0.1 mg/mL) and J. acutus (IC₅₀ = 0.4 mg/mL), and the ether extracts of J. acutus (IC₅₀ = 0.2 mg/mL) and A. macrostachyum (IC₅₀ = 0.3 mg/mL). The highest radical scavenging activity (RSA) against the 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical was obtained in the ether extract of J. acutus (IC₅₀ = 0.4 mg/mL) and H. portulacoides (IC₅₀ = 0.9 mg/mL). The maximum total phenolic content (TPC) was found in the methanol extract of M. edule (147 mg gallic acid equivalents (GAE)/g) and in the ether extract of J. acutus (94 mg GAE/g). Significant decreases in nitric oxide (NO) production were observed after incubation of macrophages with lipopolysaccharide (LPS) and the chloroform extract of H. portulacoides (IC₅₀ = 109 µg/mL) and the hexane extract of P. coronopus (IC₅₀ = 98.0 µg/mL). High in vitro cytotoxic activity and selectivity was obtained with the ether extract of J. acutus. Juncunol was identified as the active compound and for the first time was shown to display selective in vitro cytotoxicity towards various human cancer cells.     

茜草不同极性提取物的体外抑菌活性研究

本文采用k-b纸片法研究了茜草不同极性溶剂提取物的抑菌作用,结果表明,石油醚提取物浓度为50mg／ml时,抑菌率为25．03％;乙酸乙酯层提取物浓度为50mg／mL时抑制率最高,为48．64％;丙酮层提取物浓度为50mg／mL时抑制率最高,为61．59％;甲醇层提取物浓度为50mg／mL时抑制率最高,为46．09％。     

Free radical scavenging activity of selected medicinal plants of Eastern Botswana

Water and methanol extracts from roots of Ozoroa paniculosa (Anarcardiaceae); seeds of Colophospermum mopane (Caesalpiniaceae) and Cucumis metuliferus (Cucurbitaceae) ripe fruits were assessed for in vitro antioxidant activity. Free radical scavenging activity was measured spectrophotometrically as maximum fading power of DPPH at 525 nm. Water and methanol extracts of Ozoroa paniculosa exhibited higher scavenging potency than extracts of either Colophospermum mopane or Cucumis metuliferus at all tested concentrations. None of the extracts from Cucumis metuliferus exhibited any recognizable free radical scavenging activity. Above 50 microg mL(-1) both water and methanol extracts of Ozoroa paniculosa exhibited 91% scavenging activity similar to the control compounds L-ascorbic acid (91%) and (-) epicatechin (92%). Between 50-100 microg mL(-1), water and methanol extracts of Colophospermum mopane exhibited scavenging potency of < or = 70%. However, above 100 microg mL(-1), both water and methanolic extracts of C. mopane exhibited scavenging activity > 70%. Chloroform extracts of all the tested plants showed poor scavenging activity (< 30%). The order of scavenging potency for the tested samples was as follows: L-ascorbic acid > or = epicatechin > O. paniculosa (methanolic extract) > O. paniculosa (water extract) > O. paniculosa (ethylacetate extract) > C. mopane (methanolic extract) > C. mopane (water extract) > all extracts of C. metuliferus. These findings lend credence to the use of these plants as anti-inflammatory and antioxidant agents in folk medicine.     

Effect of Extraction Temperature on Pressurized Liquid Extraction of Bioactive Compounds from Fucus vesiculosus

This study was aimed at investigating the effect of low polarity water (LPW) on the extraction of bioactive compounds from Fucus vesiculosus and to examine the influence of temperature on the extraction yield, total phenolic content, crude alginate, fucoidan content, and antioxidant activity. The extractions were performed at the temperature range of 120–200 °C with 10 °C increments, and the extraction yield increased linearly with the increasing extraction temperature, with the highest yields at 170–200 °C and with the maximum extraction yield (25.99 ± 2.22%) at 190 °C. The total phenolic content also increased with increasing temperature. The extracts showed a high antioxidant activity, measured with DPPH (2,2-Diphenyl-1-picrylhydrazyl) radicals scavenging and metal-chelating activities of 0.14 mg/mL and 1.39 mg/mL, respectively. The highest yield of alginate and crude fucoidan were found at 140 °C and 160 °C, respectively. The alginate and crude fucoidan contents of the extract were 2.13% and 22.3%, respectively. This study showed that the extraction of bioactive compounds from seaweed could be selectively maximized by controlling the polarity of an environmentally friendly solvent.     

Antioxidant, antibacterial, and antiviral effects of Lactuca sativa extracts

Antioxidant, antibacterial and antiviral effects of aqueous and methanol extracts of Lactuca sativa var longifolia leaves were investigated. The antioxidant activity was evaluated using the DPPH assay. The effect of the extracts against 5 Gram-positive and 6 Gram-negative bacteria was tested. The antiviral activity was determined against human cytomegalovirus (HCMV) strain AD-169 (ATCC Ref. VR 538) and coxsackie B virus type 3 (CoxB-3) using a cytopathic effect (CPE) reduction assay. The methanol extract had the highest total phenolic contents (235.31 mg CE/g extract). It exhibited a significantly (p < 0.05) greater hydroxyl radical-scavenging activity (IC₅₀ = 3.5 μg/ml) than the aqueous extract (4.1 μg/ml). It was also the most effective extract with the lowest MIC (2.5 mg/ml) against all Gram negative and Gram positive bacteria. Methanol and aqueous extracts exhibited antiviral activity against HCMV and Cox-B3 viruses with IC₅₀ of 200 μg/ml.     

Enzyme inhibitory and antioxidant activity of Melia azedarach L. naturalized in Anatolia and its phenolic acid and fatty acid composition

In the present study, acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and tyrosinase inhibitory and antioxidant activities of the fruit and leaf extracts of Melia azedarach L. (Meliaceae) of Turkish origin were evaluated. Enzyme inhibitory activity of the extracts was tested in vitro using ELISA microplate reader. Antioxidant activity of the extracts was tested using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferrous ion-chelation, and ferric-reducing antioxidant power (FRAP) assays. Phenolic composition of the extracts was elucidated by high performance liquid chromatography (HPLC) and fatty acid compositions of the fatty oils of the fruits and leaves were elucidated by gas chromatography-mass spectrometry (GC-MS). The ethyl acetate extract from the leaves showed the highest inhibition against AChE (33.63 ± 1.40%) and BChE (92.89 ± 3.05%). The methanol extract from the leaves exerted the best antioxidant activity in DPPH radical scavenging and FRAP assays, while the ethyl acetate extracts of the fruits and leaves had the most notable effect in metal-chelation assay.     

地胆头化学成分及其生理活性的研究

研究地胆头提取液中总多酚和类黄酮含量及其生理活性。测定了不同地胆头提取液的总多酚含量、类黄酮含量及其清除DPPH自由基的能力,并比较了不同料液比的地胆头提取液对血管紧张素转化酶(ACE)的抑制率。结果表明:不同提取剂提取的地胆头总多酚和类黄酮含量不同,其中,丙酮提取液中总多酚和类黄酮含量分别为12.27 mg/g、7.50 mg/g,均高于乙醇、甲醇、水提取液中的总多酚和类黄酮含量;地胆头丙酮、甲醇、乙醇提取液对DPPH自由基的清除率分别为94.60%、93.41%、90.87%,均高于人工合成氧化剂BHT(89.15%)和地胆头水提取液(62.75%);地胆头提取液对ACE的抑制效果随着料液比的增加而越明显,其中,1∶10的料液比下,乙醇地胆头提取液对ACE抑制率达到96.67%。通过本研究得出,在地胆头乙醇、甲醇、水、丙酮4种提取液中,丙酮提取液的总多酚和类黄酮含量最高,且其抗氧化性效果最佳,乙醇地胆头提取液的抗高血压能力最强。地胆头提取液具有明显的抗氧化性和抗高血压等生理活性,在功能食品及中草药领域里具有良好的开发应用前景。     

Bioactive properties and chemical constituents of methanolic extract and its fractions from Jatropha curcas oil

The present work is designed to evaluate the bioactive properties of the crude methanolic extract of Jatropha curcas oil and its solvent fractions. The crude methanolic extract obtained was fractionated using a hydrophilic lipophilic balanced (HLB) cartridge and then eluted with different solvents in the order of hexane (F1), dichloromethane (F2), chloroform (F3), ethyl acetate (F4) and methanol (F5), respectively. Total phenolic content of the crude methanolic extract and its fractions was in the range of 0.19-4.5 mg/g as gallic acid equivalent. Antioxidant activity of the crude methanolic extract and its fractions were determined by two complementary test methods, namely, phenanthroline method and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging method. All samples demonstrated weak antioxidant activity (150-851 μmol Fe/100 g of the extract and IC₅₀ of 1.05-13.5 mg/mL). When compared to butylated hydroxytoluene (BHT), a reference synthetic antioxidant, both showed weaker antioxidative potential. The evaluation of antimicrobial activity of the extracts was performed using a disc diffusion method and a micro-well dilution method against six economic plant disease bacteria. The results showed that all extracts possessed strong to moderate antibacterial activity with varying degrees of growth inhibition against the test bacteria. The minimum inhibitory concentrations (MIC) were in the range of 14.92-428.6 μg/mL. In addition, the chemical constituents in each fraction of the extract were subjected to analyze by gas chromatography-mass spectrometry (GC-MS). The eleven constituents were identified. Among them, 2,4-di-tert-butylphenol, methyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionate and linoleic acid may be the main cause of its strong antibacterial activity. Therefore, this oil present in the methanolic extract had great potential as effective antibacterial sources.     

椰子纳塔培养液的抗氧化活性

以新鲜和发酵后椰子纳塔培养液提取物为试验材料,在测定提取物总酚含量的基础上(总酚含量新鲜纳塔培养液提取物1.86mg/mL±0.01 mg/mL,发酵纳塔培养液提取物1.36mg/mL±0.01 mg/mL),研究了2种培养液提取物对脂质过氧化的抑制作用、DPPH自由基和羟基自由基的清除作用、Fe~(2+)的络合能力以及Fe~(3+)的还原能力,同时以BHT、没食子酸为对照.结果表明,2种培养液提取物均具有明显的抗氧化性.新鲜培养液对脂质过氧化的抑制作用、DPPH自由基和羟基自由基清除作用、Fe~(2+)络合能力、Fe~(3+)的还原能力均高于发酵后培养液提取物的其作用.     

Characterization of Protein Hydrolysates from Fish Discards and By-Products from the North-West Spain Fishing Fleet as Potential Sources of Bioactive Peptides

Fish discards and by-products can be transformed into high value-added products such as fish protein hydrolysates (FPH) containing bioactive peptides. Protein hydrolysates were prepared from different parts (whole fish, skin and head) of several discarded species of the North-West Spain fishing fleet using Alcalase. All hydrolysates had moisture and ash contents lower than 10% and 15%, respectively. The fat content of FPH varied between 1.5% and 9.4% and had high protein content (69.8–76.6%). The amino acids profiles of FPH are quite similar and the most abundant amino acids were glutamic and aspartic acids. All FPH exhibited antioxidant activity and those obtained from Atlantic horse mackerel heads presented the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, reducing power and Cu²⁺ chelating activity. On the other hand, hydrolysates from gurnard heads showed the highest ABTS radical scavenging activity and Fe²⁺ chelating activity. In what concerns the α-amylase inhibitory activity, the IC₅₀ values recorded for FPH ranged between 5.70 and 84.37 mg/mL for blue whiting heads and whole Atlantic horse mackerel, respectively. α-Glucosidase inhibitory activity of FPH was relatively low but all FPH had high Angiotensin Converting Enzyme (ACE) inhibitory activity. Considering the biological activities, these FPH are potential natural additives for functional foods or nutraceuticals.     

几种野生稻甲醇提取物对害虫的行为干扰及对酯酶同工酶的影响

采用索氏回流法,经甲醇提取,获得大颖野生稻（E6 3/6 4、E6 1）、小粒野生稻（E13 9、E13 13）、药用野生稻（E15 8、E15 13）、斑点野生稻（E16 1、E16 3、E16 13）、阔叶野生稻（101392、E9 1、E9 10）和颗粒野生稻（E7 4）甲醇提取物。室内生物活性测定表明这些野生稻甲醇提取物对柑橘全爪螨和绣线菊蚜具有一定的忌避活性,其中药用野生稻E15 8的甲醇抽提物效果更为显著,5×104 μg/mL的浓度处理,测得24 h和48 h对柑橘全爪螨的忌避率分别为83.26%和87.95%;用同样的浓度处理,对绣线菊蚜的忌避率分别为87.86%和82.43%。经聚丙烯酰胺凝胶电泳检测,药用野生稻E15 8的甲醇抽提物对两种害虫的酯酶同工酶具有明显的抑制效果。     

Effect of Boiling in Water of Barley and Buckwheat Groats on the Antioxidant Properties and Dietary Fiber Composition

In recent years, there has been an ever-increasing interest in the research of polyphenols obtained from dietary sources, and their antioxidative properties. The purpose of this study was to determine the effect of boiling buckwheat and barley groats on the antioxidant properties and dietary fiber composition. Antioxidative properties were investigated using methyl linoleate model system, by assessing the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity and metal chelating activity. The results were compared with butylated hydroxytoluene (BHT). Raw barley and buckwheat groats extracts showed higher DPPH scavenging ability compared to boiled barley and buckwheat groats extracts. Raw barley groats extract exhibited higher antioxidant activity than boiled groats extract in the methyl linoleate emulsion. Higher chelating ability in relation to Fe (II) ions was observed for boiled groats extracts as compared to raw groats extracts. BHT showed small antiradical activity and metal chelating activity, while showing higher antioxidative activity in emulsion system. The analysis of groats extracts using HPLC method showed the presence of rutin, catechin, quercetin, gallic, p-hydroxybenzoic, p-coumaric, o-coumaric, vanillic, sinapic, and ferulic acids. Differences in the content of dietary fiber and its fractions were observed in the examined products. The highest total dietary fiber content was detected in boiled buckwheat groats, while the lowest - in boiled barley groats. The scientific achievements of this research could help consumers to choose those cereal products available on the market, such as barley and buckwheat groats, which are a rich source of antioxidative compounds and dietary fiber.     

Comparative Study of Onion (Allium cepa) and Leek (Allium ampeloprasum): Identification of Organosulphur Compounds by UPLC-QTOF/MS and Anticancer Effect on MCF-7 Cells

Onion (Allium cepa) and leek (Allium ampeloprasum var. porrum) are common herbs and vegetables found in our daily life. It belongs to the genus Allium, which is usually known for their high antioxidant and anticancer properties. Medical researchers highly recommend the exploitation of herbs and plants as alternative ways in the treatment of cancer. This research was designed to study the anticancer effects of onion and leek extracts on MCF-7 human breast cancer cell. Crude extracts of ethanol, methanol, and water of onion and leek were obtained by maceration. MCF-7 cells were cultured in complete media at 37 °C and subjected to different treatments that involved varying concentrations (10, 50, and 100 μg/mL) of onion and leek extracts for 24, 48, and 72 h of incubation. The percentage of cell viability and the concentration of extracts on MCF-7 cells were determined using MTT assay. The water leek extract proved to be the most effective extract at 50 μg/mL, whereby it showed a significant inhibition ability due to the presence of entadamide A-β-D-glucopyranoside as identified by ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (UPLC-QTOF/MS). Further studies about the mechanism of both extracts in causing cell death and the determination of the presence of other bioactive compounds in the extracts are needed.

     

Comparative assessment of antioxidant and cholinesterase inhibitory properties of the marigold extracts from Calendula arvensis L. and Calendula officinalis L.

Calendula sp. is an important medicinal and industrial plant with various bioactivities. In this study, we examined enzyme inhibitory effects of the n-hexane, dichloromethane, acetone, ethyl acetate, methanol, and water extracts of the leaf and flowers of Calendula arvensis L. and C. officinalis L. against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The extracts were screened for their antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric ion-chelating capacity and ferric-reducing antioxidant power (FRAP) assays at 250, 500, and 1000 μg mL⁻¹. Total phenol and flavonoid quantification of the extracts was achieved using Folin-Ciocalteau and AlCl₃ reagents, respectively. The ethyl acetate extract of C. arvensis flowers was the most active in AChE inhibition assay (31.24 ± 1.29%), while the n-hexane extract of C. officinalis leaves exerted the highest ferric ion-chelating capacity (74.27 ± 2.25%). Thin layer chromatographic analysis indicated presence of flavonoid and triterpene derivatives mainly in the extracts.     

Anti-Proliferative Properties,Biocompatibility, and Chemical Composition of Different Extracts of Plantago major Medicinal Plant

Background:To study the anticancer activity of Plantago major, we assessed the effect of ethanolic, methanolic and acetonic extracts of this plant on HCT-116, SW-480, and HEK-293 cell lines as control. Methods:The cytotoxic activity, biocompatibility, and toxicity were evaluated by MTT assay, hemolysis, and Artemia salina-LD₅₀ (on mice) tests, respectively. The analysis of the extracts was performed by GC-MS analysis. Results:The results showed that all the extracts had the most antiproliferative properties on the HCT-116 cell line. The P. major root extract was more effective than the aerial parts, and IC₅₀ values for ethanolic, methanolic and acetonic root extracts were 405.59, 470.16, and 82.26 µg/mL, respectively on HCT-116 cell line at 72 h. Hemolysis degree of the ethanolic extract of aerial and root parts were approximately 1% at 400 μg/mL.. Using the ethanolic extracts, the Artemia survived every concentration, and no toxicity was observed. One week after the oral administration of different parts of P. major extracts, none of the mice died, even those were administered 2000 mg/kg. The results of GC/MS analysis showed that P. major extracts contain potential anticancer compounds, such as stearic acid (8.61%) in aerial parts of methanolic extract and 1,2- Benzenedicarboxylic acid, mono(2-ethylhexyl)ester (88.07% and 40.63%) in aerial and root parts of acetonic extract of P. major. Conclusions:Our findings suggest that the P. major is a source of potential compounds with antiproliferative properties. Key Words: Gas chromatography-mass spectrometry, HCT-116 cells, Hemolysis, Lethal dose 50     

Inhibitory Potential of Acroptilon repens against Key Enzymes involved in Alzheimer and Diabetes,Phytochemical Profile,Radical Scavenging,and Antibacterial Activity

Background:This study was devoted to assessing the inhibitory potential of acetone, methanol, and ethanol extracts of Acroptilon repens against disease-associated enzymes, as well as their antioxidant/antibacterial activity and phytochemical composition. Methods:Comparative assessment using various antioxidant evaluation methods, including FRAP, scavenging ability on DPPH radical and hydrogen peroxide, and RP, indicated that the acetone extract presented the highest antioxidant activity, due to its highest total antioxidant content. Results:The TPC and TFC of these extracts were 3.44 ± 0.32 mg GAE/g DW and 2.09 ± 0.2 mg QE/g DW, respectively. The hydrodistillation essential oil from A. repens was analyzed by GC/MS, and 17 compounds were identified. All extracts showed good inhibitory activities against disease-related enzyme acetylcholinesterase and α-amylase, with the lowest IC₅₀ for acetonic extract. Extracts of A. repens exhibited inhibiting activities against the Gram-positive bacteria, with the most effect of acetone extract. Conclusion:Our findings suggest A. repens as a promising source of natural antioxidant, antimicrobial, anti-cholinesterase and anti-amylase agents for the management of oxidative damage, and pharmaceutical, food, and cosmeceutical purposes. Key Words: Acroptilon repens, Antioxidants, Phytochemicals     

Chemical Composition and Antioxidant/Antimicrobial Activities in Supercritical Carbon Dioxide Fluid Extract of Gloiopeltis tenax

Gloiopeltis tenax (G. tenax) is widely distributed along the Chinese coastal areas and is commonly used in the treatment of diarrhea and colitis. This study aimed at investigating the bioactivities of the volatile constituents in G. tenax. We extracted the essential constituents of G. tenax by supercritical carbon dioxide extraction (CO₂-SFE), then identified and analyzed the constituents by gas chromatography-mass spectrometry (GC-MS). In total, 30 components were identified in the G. tenax extract. The components showed remarkable antioxidant activity (radical scavenging activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH)), lipid peroxidation inhibition capacity (in a β-carotene/linoleic acid-coupled oxidation reaction), and hydroxyl radical-scavenging activity (by deoxyribose degradation by iron-dependent hydroxyl radical), compared to butylated hydroxytoluene. In microdilution assays, G. tenax extracts showed a moderate inhibitory effects on Staphyloccocus aureus (minimum inhibitory concentration (MIC) = 3.9 mg/mL), Enterococcus faecalis (7.8 mg/mL), Pseudomonas aeruginosa (15.6 mg/mL), and Escherichia coli (3.9 mg/mL). Antioxidant and antimicrobial activities of G. tenax were related to the active chemical composition. These results suggest that the CO₂-SFE extract from G. tenax has potential to be used as a natural antioxidant and antimicrobial agent in food processing.     

Potato disc bioassay and cytotoxic effect of Leptadenia pyrotechnica: comparative study of diverse extracts

Comparative acute toxicity studies of the latex and sequential extracts of Leptadenia pyrotechnica (Forsk.) Decne (Asclepiadaceae) were recorded using brine shrimp. The higher toxicities were exhibited in latex; methanol, methanol/dichloromethane (1:1), defatted methanol/dichloromethane (1:1), defatted methanol and dichloromethane extracts. The other extracts; aqueous, alkaloids, ethyl acetate and n-butanol exhibited less toxicities compared with the other extracts. The estimated LC50 and its 95% confidence limits for these extracts expressed in ppm were: methanol, latex 18.84 (11.22-31.61), methanol/dichloromethane 19.95 (7.76-53.70), defatted methanol/dichloromethane 21.38 (7.24-63.10), defatted methanol 28.19 (16.27-48.81) and dichloromethane 30.90 (11.75-79.43). The anti-tumor activities; potato disc assays of methanol, ethyl acetate and alkaloids extracts showed good activities as anti-tumor agent which represented-49.30,-43.20 an -33.60%, respectively. While latex and aqueous extract represented-30.80 and-28.17%, respectively.     

Antioxidant Activity and Phenolic Content of Betalain Extracts from Intact Plants and Hairy Root Cultures of the Red Beetroot Beta vulgaris cv. Detroit Dark Red

Betalains are water-soluble plant pigments that are widely used as food colorants, and have a wide range of desirable biological activities, including antioxidant, anti-inflammatory, hepatoprotective, anti-cancer properties. They can be produced from various plants, notably beetroot, but betalain products obtained in this way also have some undesirable properties and are difficult to standardize. A potentially attractive alternative is to use hairy root cultures. In the study reported here, we found that betalain extracts obtained from hairy root cultures of the red beetroot B. vulgaris cv. Detroit Dark Red also had higher antioxidant activity than extracts obtained from mature beetroots: six-fold higher 2,2-dyphenyl-1-picrylhydrazyl radical scavenging ability (90.7% inhibition, EC₅₀ = 0.11 mg, vs 14.2% inhibition, EC₅₀ = 0.70 mg) and 3.28-fold higher oxygen radical absorbance capacity (4,100 μM TE/g dry extract, vs 1,250 μM TE/g dry extract). The high antioxidant activity of the hairy root extracts was associated with increased concentrations (more than 20-fold) of total phenolic concomitant compounds, which may have synergistic effects with betalains. The presence of 4-hydroxybenzoic acid, caffeic acid, catechin hydrate, and epicatechin were detected in both types of extract, but at different concentrations. Rutin was only present at high concentration (1.096 mg.g⁻¹ dry extract) in betalain extracts from the hairy root cultures, whereas chlorogenic acid was only detected at measurable concentrations in extracts from intact plants.