Luteolin Glucuronides Research Articles

Mentha piperita leaf extract suppresses the release of ATP from epidermal keratinocytes and reduces dermal thinning as well as wrinkle formation.

To achieve a more beautiful and younger appearance, reducing wrinkles is a key concern. The process of wrinkle formation is complex and the development of truly effective cosmetic ingredients to reduce wrinkles remains a challenge. Recent studies have revealed a close relationship between wrinkles and skin thinning, suggesting that preventing skin thinning could also prevent wrinkle formation. In this study, we examined the role of extracellular adenosine triphosphate (eATP) in the progression of thinning, as eATP reportedly increases skin ageing factors, such as senescence-associated secreted phenotype (SASP) factors in epidermal cells. We determined the effects of Mentha piperita leaf extract on suppressing eATP to reduce thinning and wrinkles. Adenosine triphosphate (ATP) levels were measured in normal human epidermal keratinocytes (NHEK) in the presence of M. piperita leaf extract. Dryness, high pH, and UVB radiation were used as extrinsic ageing factors. Intrinsic skin ageing was evaluated by comparing cells from adults (AD-NHEK) and newborns (NB-NHEK). A placebo-controlled invivo study was carried out with a formulation containing 1% M. piperita leaf extract. The eATP levels were significantly higher in AD-NHEK compared with that in NB-NHEK cells. M. piperita leaf extract significantly decreased eATP levels in adult cells. Extrinsic ageing factors increased eATP levels in NHEK, whereas M. piperita leaf extract significantly suppressed eATP under all conditions. The active components of M. piperita leaf extract, luteolin glucuronide and rosmarinic acid, also decreased eATP. Moreover, compared with placebo lotion, M. piperita leaf extract-formulated lotion markedly increased dermal thickness and reduced wrinkles associated with crow's feet and the neck area. We demonstrated for the first time that M. piperita leaf extract containing rosmarinic acid and luteolin-7-O-glucuronide has the potential to reduce eATP release from epidermal keratinocytes. An increase in eATP was observed not only during inflammation but also during natural ageing. Furthermore, the invivo experiment revealing that 1% M. piperita leaf extract-containing lotion improved dermal thinning and wrinkles across multiple areas is attributed to the amelioration of dermal thinning. Thus, our data suggest the possibility of a novel cosmetic approach for reducing skin ageing by reducing eATP-mediated dermal thinning.

A comprehensive review of phytochemistry and antibacterial action of Tectona grandis

Tectona grandis L. (Teak) is one of the commercial plants which is often used in Southeast Asia. Globally, the total area of planted teaks reaches 3 million hectares, with about 31% of the land located in Indonesia. Preliminary phytochemistry showed bioactive constituents as alkaloids, tannins, flavonoids and phenolic compounds. Isolation and characterization by several methods were reported for quantifying the secondary metabolites found in the various parts of Tectona G, following the ICH guidelines and the following were reported, viz; Protocatechuic acid, Quinic acid, and its derivatives, Apigenin 7-O-diglucuronide, Luteolin, Luteolin 7-Odiglucuronide, Luteolin glucuronide, Diglucuronide, Apigenin glucuronide etc, which are responsible for their antibacterial action. The extractives of Tectona grandis were studied for antibacterial activities by agar well diffusion and disc diffusion method respectively. The test organism were isolates of nutrient broth, hospital strains, decayed food and Mueller Hinton Broth (MHB), antibacterial action of Tectona G were lucid as the extractives showed a maximum inhibitory effect; inhibition zone, evident by a clear zone of inhibition around the discs, minimum inhibitory concentration (MIC) and minimum bacteriocidal concentration (MBC) respectively. Accordingly, this review demonstrates the importance of the herbal plant (Tectona grandis) in the treatment of bacteria thus, preventing its resistance.

A Multi-Approach Study of Phytochemicals and Their Effects on Oxidative Stress and Enzymatic Activity of Essential Oil and Crude Extracts of Rosmarinus officinalis

Rosmarinus officinalis or Rosemary is a highly valued medicinal vegetal, owing to its notable antispasmodic, anti-inflammatory, and antibacterial properties. In the current work, we aimed to identify the chemical components of the essential oil (EO) of R. officinalis and evaluate its biological properties using an in vitro approach. High performance liquid chromatography time-of-flight mass spectrometry (HPLC-TOF-MS) was utilized to analyze of the hydro-methanolic extract (HME), while gas chromatography–mass spectrometry (GC/MS) was considered during the analysis of the EO’s chemical composition. The antioxidant abilities of HME and the EO were assessed using diverse tests (DPPH, ABTS, GOR, CUPRAC, and FRAP). The anti-enzymatic properties were tested by the inhibition of cholinesterases, α-glucosidase, and tyrosinase enzyme. The HPLC-TOF-MS displayed the existence of flavonoids like luteolin glucuronide I and II, and a few known hydroxycinnamic acids. The EO contained three major components, namely, eucalyptol (28.7%), camphor (16.7%), and borneol (13.5%). The HME had a high total polyphenol content, as determined by the Folin–Ciocalteau method (335.37 ± 9.33 µg of gallic acid eq·mg−1). Notably, the analysis of the bioactivities of the HME and EO revealed comparatively that they possessed higher radical scavenging capacity in the DPPH, ABTS, and galvinoxyl assays, while EO exhibited a higher capacity for enzyme inhibition. Overall, our findings suggest that both the EO and HME extract of Algerian’s R. officinalis holds great usefulness in the pharmaceutical and nutraceutical fields due to its elevated polyphenol content and potent bioactivities.

Effect of the main constituents of Pistacia lentiscus leaves against the DPPH radical and xanthine oxidase: experimental and theoretical study

The aim of this work is to study the content of phenolic compounds in P lentiscus leaves and their antioxidant effect. After extracting the phenolic compounds, fractionation by liquid/liquid partition with increasing polarity gives five extracts. Three of them (ButF, AqF and ButA) were found to have good antioxidant activity. Their IC50s for the inhibition of the free radical formation of DPPH are 1.76 µg/mL, 1.307 µg/ml, and 1.77 µg/mL, respectively. These values are very interesting, considering the effect of the powerful flavonoid quercetin, whose IC50 against DPPH is 1.53 µg/mL. These extracts are also active against xanthine oxidase (XO). The IC50s measured are 0.14 mg/mL, 0.186 mg/mL and 0.33 mg/mL for ButF, Aq F and ButAq F extract respectively, in comparison with allopurinol (0.44 mg/mL). A phytochemical analysis by LC/ESI-MS-MS was performed to explain the observed activities. The results show 22 peaks representing: flavanols, namely catechin, d-Gallocatechin, and gallocatechin gallate. The only flavone detected in the studied extracts was luteolin glucuronide and was found to be in higher amounts in butanolic extract (2,71mg/mL). The phenolic acids and derivatives were also identified in the extracts. A theoretical study was performed to deduce the specificity of the binding between the major compounds identified in the P. lentiscus extract and the xanthine oxidase enzyme using Schrödinger software. The docking procedure was validated using the extraction of ligands from the binding site. Their re-anchoring to the xanthine oxidase structure using quercetin and allopurinol was considered reference molecules. After docking, post-docking minimization was performed to achieve the best scoring poses with the MM-GBSA approach. The dGBind energy of MM-GBSA representing the binding energy of the receptor and the ligand was calculated based on molecular mechanics. Results reveal that β-Glucogallin compounds such as Digalloylquinic acid, Gallocatechin, and Myricetin-3-O rhamnoside are more active than allopurinol, with stronger Docking score (Gscore) and MM-GBSA dGBind. Communicated by Ramaswamy H. Sarma

Comparative biotransformation of luteolin and apigenin from the flower extract and the stem-and-leaf extract of Dendranthema morifolium Ramat. Tzvel. in rats.

The flower of Dendranthema morifolium Ramat Tzvel has been widely used as a nutritional health supplement worldwide. However, most of the studies have focused on the flower and the rest of the plant was neglected. Our hypothesis is that similar flavonoids may be present at different parts of D. morifolium, and the flavonoids may undergo a similar biotransformation pathway within the body. To investigate this hypothesis, an in vivo pharmacokinetic experimental model was developed to explore the comparative biotransformation of luteolin and apigenin after administration of D. morifolium extracts (10g kg-1 , p.o.) in freely moving rats. Because luteolin and apigenin mainly underwent phase II metabolism, the metabolic enzymes of β-glucuronidase/sulfatase or β-glucuronidase were used to hydrolyze the plasma sample, depending on the biotransformation pathway involved. The results revealed that luteolin and apigenin mainly went through glucuronide and sulfate conjugations, respectively, in both the extract of flowers and the stem-and-leaf group. In addition, the area under the concentration curve (AUClast ) of luteolin glucuronides and sulfates in the group administered the stem-and-leaf extract was approximately 4.6 times higher than that of the flower extract group. The dominant products of biotransformation for apigenin were sulfates. These findings support our hypothesis that not only the flower parts of D. morifolium, but also the stem-and-leaf parts contain rich flavones, including glycosides and aglycone, and they undergo similar biotransformation pathways. © 2021 Society of Chemical Industry.

Importance of OATP1B1 and 1B3 in the Liver Uptake of Luteolin and Its Consequent Glucuronidation Metabolites

Luteolin is a typical flavonoid and broadly distributed in the plants. Oral bioavailability of luteolin is low owing to extensive metabolism. Regioselective glucuronidation by UDP-glucuronosyltransferases (UGTs) and liver uptake by organic anion transporting polypeptides (OATPs) of luteolin and consequent glucuronidation metabolites were studied. Luteolin-3'-O-glucuronide (L-3'-G) and luteolin-7-O-glucuronide (L-7-G) were the major metabolites in human liver microsomes. Further study demonstrated that UGT1A9 played a predominant role in the glucuronidation of luteolin. Transporter study showed that OATP1B1- and 1B3-transfected cells selectively uptake L-3'-G into cells but not luteolin or L-7-G. After intravenous administration of luteolin to mice, the area under the curve of L-3'-G in the plasma was the highest among luteolin, L-3'-G, and L-7-G. In the liver, the concentration of L-3'-G was significantly greater than L-7-G. In conclusion, OATP1B1 and OATP1B3 play an important role in the liver disposition of luteolin and its glucuronidation metabolites.

Assessment of selected Saudi and Yemeni plants for mosquitocidal activities against the yellow fever mosquito Aedes aegypti

As part of our continuing investigation for interesting biological activities of native medicinal plants, thirty-nine plants, obtained from diverse areas in Saudi Arabia and Yemen, were screened for insecticidal activity against yellow fever mosquito Aedes aegypti (L.). Out of the 57 organic extracts, Saussurea lappa, Ocimum tenuiflorum, Taraxacum officinale, Nigella sativa, and Hyssopus officinalis exhibited over 80% mortality against adult female Ae. aegypti at 5 μg/mosquito. In the larvicidal bioassay, the petroleum ether extract of Aloe perryi flowers showed 100% mortality at 31.25 ppm against 1st instar Ae. aegypti larvae. The ethanol extract of Saussurea lappa roots was the second most active displaying 100% mortality at 125 and 62.5 ppm. Polar active extracts were processed using LC-MS/MS to identify bioactive compounds. The apolar A. perryi flower extract was analyzed by headspace SPME-GC/MS analysis. Careful examination of the mass spectra and detailed interpretation of the fragmentation pattern allowed the identification of various biologically active secondary metabolites. Some compounds such as caffeic and quinic acid and their glycosides were detected in most of the analyzed fractions. Additionally, luteolin, luteolin glucoside, luteolin glucuronide and diglucuronide were also identified as bioactive compounds in several HPLC fractions. The volatile ketone, 6-methyl-5-hepten-2-one was identified from A. perryi petroleum ether fraction as a major compound.

Infusions and decoctions of dehydrated fruits of Actinidia arguta and Actinidia deliciosa: Bioactivity, radical scavenging activity and effects on cells viability

Actinidia deliciosa and A. arguta fruits (kiwifruit and kiwiberry, respectively) are an excellent source of bioactive compounds. The aim of this paper is to valorize the fruits that are not commercialized (e.g. due to inadequate size or physical damage) in infusions and decoctions. The antioxidant activity, the scavenging activity against reactive species, the phenolic profile and the intestinal effects of infusions and decoctions of dehydrated fruits were evaluated and compared. Decoctions presented the highest antioxidant activity and a good ability to capture HOCl and NO. The phenolic composition of A. arguta present quinic acid, cis-caftaric acid and its derivatives, caffeoyl hexoside, luteolin glucuronide, quercetin derivatives and myristin, while A. deliciosa extracts were characterized by the presence of quinic acid, caffeic acid and its derivatives and caffeoyl hexoside. No adverse effects were observed on Caco-2 and HT29-MTX cells. Kiwiberry decoctions showed to be the best option to keep the fruits benefits.

Absorption and Metabolism of Luteolin in Rats and Humans in Relation to in Vitro Anti-inflammatory Effects

Luteolin is a flavonoid present in plants in the form of aglycone or glucosides. In this study, luteolin glucosides (i.e., luteolin-7- O-β-d-glucoside, luteolin-7- O-[2-(β-d-apiosyl)-β-d-glucoside], and luteolin-7- O-[2-(β-d-apiosyl)-6-malonyl-β-d-glucoside]) prepared from green pepper leaves as well as luteolin aglycone were orally administered to rats. Regardless of the administered luteolin form, luteolin glucuronides were mainly detected from plasma and organs. Subsequently, luteolin aglycone, the most absorbed form of luteolin in rats, was orally administered to humans. As a result, luteolin-3'- O-sulfate was mainly identified from plasma, suggesting that not only luteolin form but also animal species affect the absorption and metabolism of luteolin. When LPS-treated RAW264.7 cells were treated with luteolin glucuronides and luteolin sulfate (the characteristic metabolites identified from rats and humans, respectively), the different luteolin conjugates were metabolized in different ways, suggesting that such difference in metabolism results in their difference in anti-inflammatory effects.

Comparison of phenolic acids and flavonoids contents in various cultivars and parts of common lavender (Lavandula angustifolia) derived from Poland

The aim of study was to compare the content of phenolic acids and flavonoids in two cultivars of Lavandula angustifolia: ‘Blue River’ and ‘Ellagance Purple’, including flowers and leafy stalks. Total phenolics and total flavonoids contents were determined by UV–Vis spectroscopy. The contents of total phenolics in leafy stalks (3.71–4.06 mg g−1 d.m.) were higher than in flowers (1.13–1.14 mg g−1 d.m.). Similarly, higher total contents of flavonoids were determined in leafy stalks (3.41–3.51 mg g−1 d.m.), as compared with flowers (0.86–0.91 mg g−1 d.m.). Phenolic acids and flavonoids were identified and quantified using HPLC and UPLC methods. Three phenolic acids were determined: rosmarinic, ferulic and caffeic acid. Lavender extracts contained also flavonoids from group of apigenin, luteolin and quercetin. Higher amounts of luteolin diglucuronide and luteolin glucuronide were found in leafy stalks in comparison to flowers. Obtained results indicate that leafy stalks of lavender can be also valuable source of antioxidant compounds.

Metabolic Disposition of Luteolin Is Mediated by the Interplay of UDP-Glucuronosyltransferases and Catechol-O-Methyltransferases in Rats.

Luteolin partially exerts its biologic effects via its metabolites catalyzed by UDP-glucuronosyltransferases (UGTs) and catechol-O-methyltransferases (COMTs). However, the interplay of UGTs and COMTs in mediating luteolin disposition has not been well clarified. In this study, we investigated the glucuronidation and methylation pathways of luteolin mediated by the interplay of UGTs and COMTs in vivo and in vitro. A total of nine luteolin metabolites was detected in rat plasma and bile by liquid chromatography-tandem mass spectrometry, namely, three glucuronides, two methylated metabolites, and four methylated glucuronides. Luteolin-3'-glucuronide (Lut-3'-G) exhibited the highest systemic exposure among these metabolites. Kinetics studies in rat liver S9 fractions suggested two pathways, as follows: 1) Luteolin was glucuronidated to luteolin-7-glucuronide, luteolin-4'-glucuronide, and Lut-3'-G by UGTs, and then Lut-7-G was methylated to chrysoeriol-7-glucuronide and diosmetin-7-glucuronide by COMTs. 2) Alternatively, luteolin was methylated to chrysoeriol and diosmetin by COMTs, and then chrysoeriol and diosmetin were glucuronidated by UGTs to their respective glucuronides. The methylation rate of luteolin was significantly increased by the absence of glucuronidation, whereas the glucuronidation rate was increased by the absence of methylation, but to a lesser extent. In conclusion, two pathways mediated by the interplay of UGTs and COMTs are probably involved in the metabolic disposition of luteolin. The glucuronidation and methylation of luteolin compensate for each other, although glucuronidation is the predominant pathway.

Lemon balm extract (Melissa officinalis, L.) promotes melanogenesis and prevents UVB-induced oxidative stress and DNA damage in a skin cell model

BackgroundSolar ultraviolet (UV) radiation is one of the main causes of a variety of cutaneous disorders, including photoaging and skin cancer. Its UVB component (280–315nm) leads to oxidative stress and causes inflammation, DNA damage, p53 induction and lipid and protein oxidation. Recently, an increase in the use of plant polyphenols with antioxidant and anti-inflammatory properties has emerged to protect human skin against the deleterious effects of sunlight. ObjectiveThis study evaluates the protective effects of lemon balm extract (LBE) (Melissa Officinalis, L) and its main phenolic compound rosmarinic acid (RA) against UVB-induced damage in human keratinocytes. MethodsThe LBE composition was determined by HPLC analysis coupled to photodiode array detector and ion trap mass spectrometry with electrospray ionization (HPLC-DAD-ESI-IT-MS/MS). Cell survival, ROS generation and DNA damage were determined upon UVB irradiation in the presence of LBE. The melanogenic capacity of LBE was also determined. ResultsRA and salvianolic acid derivatives were the major compounds, but caffeic acid and luteolin glucuronide were also found in LBE. LBE and RA significantly increased the survival of human keratinocytes upon UVB radiation, but LBE showed a stronger effect. LBE significantly decreased UVB-induced intracellular ROS production. Moreover, LBE reduced UV-induced DNA damage and the DNA damage response (DDR), which were measured as DNA strand breaks in the comet assay and histone H2AX activation, respectively. Finally, LBE promoted melanogenesis in the cell model. ConclusionsThese results suggest that LBE may be considered as a candidate for the development of oral/topical photoprotective ingredients against UVB-induced skin damage.

Metabolic Fate of Luteolin in Rats: Its Relationship to Anti-inflammatory Effect.

Luteolin is a naturally occurring flavone that reportedly has anti-inflammatory effects. Because most luteolin is conjugated following intestinal absorption, free luteolin is likely present at low levels in the body. Therefore, luteolin metabolites are presumably responsible for luteolin bioactivity. Here we confirmed that luteolin glucuronides, especially luteolin-3'-O-glucuronide, are the major metabolites found in plasma after oral administration of luteolin (aglycone) or luteolin glucoside (luteolin-7-O-glucoside) to rats. Luteolin-4'-O-glucuronide and luteolin-7-O-glucuronide were also detectable together with luteolin-3'-O-glucuronide in the liver, kidney, and small intestine. Next, we prepared these luteolin glucuronides and compared the anti-inflammatory effects of luteolin and luteolin glucuronides on gene expression in lipopolysaccharide-treated RAW264.7 cells. Luteolin glucuronides, especially luteolin-7-O-glucuronide, reduced expression of inflammatory genes in the cells, although their effects were weaker than those of luteolin. These results indicate that the active compound responsible for the anti-inflammatory effect of luteolin in vivo would be luteolin glucuronide and/or residual luteolin.

Bioactivity of herbal tea of Hungarian thyme based on the composition of volatiles and polyphenolics

Hungarian thyme (Thymus pannonicus All., Lamiaceae) is an aromatic herb used as traditional remedy, a refreshing beverage and a food aromatizer. Herbal teas, i.e., infusions, of Hungarian thyme from eight localities in Serbia were analyzed regarding their polyphenolic and volatile composition, and tested for their antioxidant and antimicrobial activity. The total polyphenolics content, determined by the Folin-Ciocalteu method, ranged from 1122.25 to 1979.93mg gallic acid/L. HPLC analysis revealed rosmarinic acid (367.42–1199.47mg/L) and luteolin glucuronides as the main polyphenolics. The volatile fractions of the infusions, analyzed by static headspace extraction coupled with GC and GC–MS analyses, contained citral, 3-octanone, 1-octen-3-ol, linalool and 1,8-cineole as the dominant constituents. The antioxidant activity of the infusions was examined through the ferric reducing antioxidant power (FRAP) (68.09–124.58mmol Fe2+/L) and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical tests (SC50 1.32–2.96μL/mL). The antimicrobial activity was tested by the broth microdilution method against standard strains of Gram(+) Staphylococcus aureus, Staphylococcus epidermidis, Bacillus subtilis and Enterococcus faecalis and Gram(−) bacteria Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa, and one strain of yeast Candida albicans. The infusions inhibited microbial growth in the tested concentration range (31.25–500.00μL/mL) and the strongest activity was exhibited against the strain of C. albicans (MIC 31.25–62.50μL/mL). The antioxidant and antimicrobial properties of the infusions were, to some extent, in correlation with the composition and content of the polyphenolic compounds, whereas the volatiles noticeably influenced the exhibited antimicrobial activity.

Breast Cancer Resistance Protein-Mediated Efflux of Luteolin Glucuronides in HeLa Cells Overexpressing UDP-Glucuronosyltransferase 1A9

UDP-glucuronosyltransferases (UGTs) are responsible for the formation of glucuronides of polyphenolic flavonoids. This study investigated the UGT1A9-mediated glucuronidation of luteolin and the kinetics of luteolin glucuronide efflux. HeLa cells overexpressing UGT1A9 (HeLa-UGT1A9) were used to determine the kinetics of breast cancer resistance protein (BCRP)-mediated transport of luteolin glucuronides. Human UGT isoforms were used to determine glucuronidation rates. UGT1A9 was found to catalyze the production of four luteolin glucuronides, including three known monoglucuronides and a novel 3', 4'-diglucuronide. Ko143, a potent specific inhibitor of BCRP, significantly inhibited efflux of luteolin monoglucuronides from HeLa1A9 cells and increased their intracellular levels in a dose-dependent manner. The formation of luteolin diglucuronide was observed when intracellular concentration of total monoglucuronides went above 0.07nM. Intracellular accumulation of diglucuronide was detected at high monoglucuronide concentrations (>0.07nM). Diglucuronide production is speculated to be a compensatory pathway for luteolin disposition.

Antioxidant properties and polyphenolics composition of common hedge hyssop (Gratiola officinalis L.)

The antioxidant potential of Gratiola officinalis was evaluated by the off-line and on-line HPLC/UV/DPPH radical scavenging assays, phytochemical composition was analyzed by LC/MS. On-line method was validated by using reference antioxidants and linear dependence was found between their concentration and radical scavenging peak area. Radical scavenging capacity (RSC) of methanol and acetone extracts expressed in their concentration required to scavenge 50% of DPPH• was 0.10% and 0.13%, respectively; the RSC in ABTS•+ assay was 1093±104 and 746±18μM of trolox equivalents in 1g, respectively. Good correlation was observed between total amount of phenolic compounds and RSC. Preliminary HPLC/UV/MS analysis revealed that the main compounds possessing antioxidant activity in the extracts might be phenylpropanoid glycosides; UPLC/UV/ESI-QTOF-MS analysis suggested 15 structures: 2,5-dihydroxy-p-benzenediacetic and caffeic acids, apigenin 6,8-di-C-β-d-glucopyranoside (vicenin-2), apigenin 8-C-α-l-arabinoside 6-C-β-d-glucoside, (shaftoside), forsythoside B, arenarioside, verbascoside (acteoside), amioside, quercetin-6-O-(2-O-acetyl-glucopyranosyl)-glucopyranoside, isoverbascoside, quercetin glucuronide, linariifolioside, methoxy luteolin-7-O-(6-O-acetyl-glucopyranosyl)-glucopyranoside, methoxy luteolin-glucuronide and luteolin glucuronide.

Determination of Polyphenols in Mentha longifolia and M. piperita Field-Grown and In Vitro Plant Samples Using UPLC-TQ-MS

Nine polyphenols in the aerial parts of Mentha longifolia have been separated by chromatographic techniques. Their structures have been confirmed by HPLC/electrospray ionization-MS/MS. The compounds identified included rosmarinic acid, salvianolic acid L, dedihydro-salvianolic acid, luteolin-glucuronide, luteolin-diglucuronide, luteolin-glucopyranosyl-rhamnopyranoside, and eriodictyol-glucopyranosyl-rhamnopyranoside. The extracts of M. longifolia and M. piperita field plants, in vitro plants, callus tissues, and cell suspension cultures were profiled, and their polyphenol composition was compared in different tissues and quantified using ultra-performance column liquid chromatography (UPLC)/triple-quadrupole-MS in the selected-ion recording detection mode. Determination of desired compounds was based on calibration curves obtained for standards, which were previously isolated from M. longifolia aerial parts. The UPLC profiles revealed considerable differences in the synthesis of secondary metabolites among samples coming from field plants, in vitro plants, callus tissues, and cell suspension cultures. Plant tissues coming from field cultivation (for both M. piperita and M. longifolia) contained several phenolic compounds (flavonoids and phenolic acids), whereas plants from in vitro conditions, callus tissues, and suspension cultures contained only a few of them. Rosmarinic acid dominated in all of these samples. These results show that under in vitro conditions, the metabolism of phenolics undergoes a fundamental change.

Purification and characterization of three neutral extracellular isoperoxidases from rye leaves

Rye ( Secale cereale L.) seedlings; contain two major flavone glucuronides, luteolin 7- O-diglucuronyl-4′- O-glucuronide (L3GlcUA) ( 1) and luteolin 7- O-diglucuronide (L2GlcUA) ( 2) in abundance in the apoplast of primary leaves; express a large number of peroxidase isoenzymes; and release H 2O 2 into the apoplast during primary leaf development. We purified and characterized three neutral extracellular peroxidase isoenzymes (rPOXs N1, N2, and N3) that can oxidize L2GlcUA as a natural substrate. The isoelectric points and molecular weights of rPOXs N1, N2, and N3 were 6.1, 7.2, and 6.3, and 42, 37, and 51 kDa, respectively. The optimum pH of the rPOXs N1, N2, and N3 were 5.5, 5.5, and 8.5, respectively, and their optimum temperatures ranged from 45 to 50 °C for all isoenzymes. rPOXs N1, N2, and N3 recognized flavonoids with 3′, 4′-OH groups as potential substrates, but not flavonoids with a glycosylated 4′-OH group or those without a 3′-OH group. The activities on phenol-type substrates were high in the order of guaiacol > catechol > o-cresol for all isoenzymes. rPOXs N1, N2, and N3 exhibited broad reactivity with endogenous hydrogen donors including luteolin glucuronides derived from the apoplast of rye primary leaves.

Identification of phenolic compounds in artichoke waste by high-performance liquid chromatography–tandem mass spectrometry

A new fast and efficient method combining liquid chromatography coupled to ionspray mass spectrometry in tandem mode with negative ion detection is described for the qualitative analysis of artichoke waste. Forty-five phenolic compounds were identified on the basis of their mass spectra in full scan mode, mass spectra in different MS–MS modes, and retention times compared with those of available reference substances. The major compounds were found to be both caffeoylquinic and dicaffeoylquinic acids, luteolin glucuronide, luteolin galactoside, quercetin, and some quercetin glycosides.

Regioselectivity of Phase II Metabolism of Luteolin and Quercetin by UDP-Glucuronosyl Transferases

The regioselectivity of phase II conjugation of flavonoids is expected to be of importance for their biological activity. In the present study, the regioselectivity of phase II biotransformation of the model flavonoids luteolin and quercetin by UDP-glucuronosyltransferases was investigated. Identification of the metabolites formed in microsomal incubations with luteolin or quercetin was done using HPLC, LC-MS, and (1)H NMR. The results obtained demonstrate the major sites for glucuronidation to be the 7-, 3-, 3'-, or 4'-hydroxyl moiety. Using these unequivocal identifications, the regioselectivity of the glucuronidation of luteolin and quercetin by microsomal samples from different origin, i.e., rat and human intestine and liver, as well as by various individual human UDP-glucuronosyltransferase isoenzymes was characterized. The results obtained reveal that regioselectivity is dependent on the model flavonoid of interest, glucuronidation of luteolin and quercetin not following the same pattern, depending on the isoenzyme of UDP-glucuronosyltransferases (UGT) involved. Human UGT1A1, UGT1A8, and UGT1A9 were shown to be especially active in conjugation of both flavonoids, whereas UGT1A4 and UGT1A10 and the isoenzymes from the UGTB family, UGT2B7 and UGT2B15, were less efficient. Due to the different regioselectivity and activity displayed by the various UDP-glucuronosyltransferases, regioselectivity and rate of flavonoid conjugation varies with species and organ. Qualitative comparison of the regioselectivities of glucuronidation obtained with human intestine and liver microsomes to those obtained with human UGT isoenzymes indicates that, in human liver, especially UGT1A9 and, in intestine, UGT1A1 and UGT1A8 are involved in glucuronidation of quercetin and luteolin. Taking into account the fact that the anti-oxidant action as well as the pro-oxidant toxicity of these catechol-type flavonoids is especially related to their 3',4'-dihydroxyl moiety, it is of interest to note that the human intestine UGT's appear to be especially effective in conjugating this 3',4' catechol unit. This would imply that upon glucuronidation along the transport across the intestinal border, the flavonoids loose a significant part of these biological activities.