Citrus × paradisi L. Fruit Waste: The Impact of Eco-Friendly Extraction Techniques on the Phytochemical and Antioxidant Potential.

Citrus fruits have been the subject of extensive research over the years due to their impressive antioxidant properties, the health benefits of flavanones, and their potential use in the prevention and treatment of chronic diseases. Grapefruit have been shown in studies to improve overall health, with numerous potential benefits, including improved heart health, reduced risk of certain cancers, improved digestive health, and improved immune system function. The development of cyclodextrin complexes is an exciting approach to increasing the content of flavanones such as naringin and naringenin in the extraction medium while improving the profile of beneficial phenolic compounds and the antioxidant profile. This research aims to optimize the extraction conditions of the flavanones naringin and naringenin with additional compounds to increase their yield from different parts of grapefruit (Citrus × paradisi L.) fruits, such as albedo and segmental membranes. In addition, the total content of phenolic compounds, flavonoids, and the antioxidant activity of ethanolic extracts produced conventionally and with -cyclodextrin was examined and compared. In addition, antioxidant activity was measured using the radical scavenging activity assay (ABTS), radical scavenging activity assay (DPPH), and ferric reducing antioxidant power (FRAP) methods. The yield of naringin increased from 10.53 ± 0.52 mg/g to 45.56 ± 5.06 mg/g to 51.11 ± 7.63 mg/g of the segmental membrane when cyclodextrins (α, ß-CD) were used; naringenin increased from 65.85 ± 10.96 µg/g to 91.19 ± 15.19 µg/g of the segmental membrane when cyclodextrins (α, ß-CD) were used. Furthermore, the results showed that cyclodextrin-assisted extraction had a significant impact in significantly increasing the yield of flavanones from grapefruit. In addition, the process was more efficient and less expensive, resulting in higher yields of flavanones with a lower concentration of ethanol and effort. This shows that cyclodextrin-assisted extraction is an excellent method for extracting valuable compounds from grapefruit.

Anti-viral and Anti-inflammatory Isoflavonoids from Ukrainian Iris aphylla Rhizomes: Structure-Activity Relationship Coupled with ChemGPS-NP Analysis.

Dried Iris rhizomes have been used in Chinese and European traditional medicine for the treatment of various diseases such as bacterial infections, cancer, and inflammation, as well as for being astringent, laxative, and diuretic agents. Eighteen phenolic compounds including some rare secondary metabolites, such as irisolidone, kikkalidone, irigenin, irisolone, germanaism B, kaempferol, and xanthone mangiferin, were isolated for the first time from Iris aphylla rhizomes. The hydroethanolic Iris aphylla extract and some of its isolated constituents showed protective effects against influenza H1N1 and enterovirus D68 and anti-inflammatory activity in human neutrophils. The promising anti-influenza effect of apigenin (13: , almost 100% inhibition at 50 µM), kaempferol (14: , 92%), and quercetin (15: , 48%) were further confirmed by neuraminidase inhibitory assay. Irisolidone (1: , almost 100% inhibition at 50 µM), kikkalidone (5: , 93%), and kaempferol (14: , 83%) showed promising anti-enterovirus D68 activity in vitro. The identified compounds were plotted using ChemGPS-NP to correlate the observed activity of the isolated phenolic compounds with the in-house database of anti-influenza and anti-enterovirus agents. Our results indicated that the hydroethanolic Iris aphylla extract and Iris phenolics hold the potential to be developed for the management of seasonal pandemics of influenza and enterovirus infections.

Fractionation and Characterization of Triterpenoids from Vaccinium vitis-idaea L. Cuticular Waxes and Their Potential as Anticancer Agents.

Fruit and leaf cuticular waxes are valuable source materials for the isolation of triterpenoids that can be applied as natural antioxidants and anticancer agents. The present study aimed at the semi-preparative fractionation of triterpenoids from cuticular wax extracts of Vaccinium vitis-idaea L. (lingonberry) leaves and fruits and the evaluation of their cytotoxic potential. Qualitative and quantitative characterization of obtained extracts and triterpenoid fractions was performed using HPLC-PDA method, followed by complementary analysis by GC-MS. For each fraction, cytotoxic activities towards the human colon adenocarcinoma cell line (HT-29), malignant melanoma cell line (IGR39), clear renal carcinoma cell line (CaKi-1), and normal endothelial cells (EC) were determined using MTT assay. Furthermore, the effect of the most promising samples on cancer spheroid growth and viability was examined. This study allowed us to confirm that particular triterpenoid mixtures from lingonberry waxes may possess stronger cytotoxic activities than crude unpurified extracts. Fractions containing triterpenoid acids plus fernenol, complexes of oleanolic:ursolic acids, and erythrodiol:uvaol were found to be the most potent therapeutic candidates in the management of cancer diseases. The specificity of cuticular wax extracts of lingonberry leaves and fruits, leading to different purity and anticancer potential of obtained counterpart fractions, was also enclosed. These findings contribute to the profitable utilization of lingonberry cuticular waxes and provide considerable insights into the anticancer effects of particular triterpenoids and pharmacological interactions.

Phenological Variations in the Content of Polyphenols and Triterpenoids in Epilobium angustifolium Herb Originating from Ukraine.

The composition of secondary metabolites undergoes significant changes in plants depending on the growth phase and the influence of environmental factors. Therefore, it is important to determine the harvesting time of plant material for the optimum secondary metabolite profile and therapeutic activity of the primary material. The shoots of Epilobium angustifolium are used as a healing tea due to the presence of polyphenolic compounds. The aim of this study was to assess the composition of phenolic compounds and triterpenoid saponins in E. angustifolium leaves and flowers and to estimate the dynamics of their content depending on the flowering phase. Qualitative and quantitative characterisation of polyphenols and triterpenoids in E. angustifolium samples from Ukraine of three flowering phases were performed using the high-performance liquid chromatography photo diode array (HPLC-PDA) method. During the present study, 13 polyphenolic compounds and seven triterpenoids were identified in the plant material. It was noted that the largest content and the best polyphenol profile was in late flowering. The most important polyphenolic compounds in the plant material were chlorogenic acid, hyperoside, isoquercitin, and oenothein B. The triterpenoid profile was at its maximum during mass flowering, with corosolic and ursolic acids being the dominant metabolites. The results of the analysis revealed that the quantity of many of the tested metabolites in the raw material of E. angustifolium is dependent on the plant organ and flowering phase. The largest content of most metabolites in the leaves was in late flowering. In the flowers, the quantity of the metabolites studied was more variable, but decreased during mass flowering and increased significantly again in late flowering. The results show that E. angustifolium raw material is a potential source of oenothein B and triterpenoids.

Distribution Patterns of Essential Oil Terpenes in Native and Invasive Solidago Species and Their Comparative Assessment.

The importance of invasive Solidago L. species to the environment creates a new approach to controlling their spread through the use of potentially high value raw materials. The aim of this study was to assess the distribution patterns of volatile compounds in the four Solidago spp., by identifying common and species-specific compounds with their potentials, and to confirm the origin of the spontaneous hybrid Solidago × niederederi on the basis of comparative assessment of essential oil (EO) profiles. Plant material in the flowering phase was collected in mixed populations from six different sites. The EOs were isolated separately from the leaf and the inflorescence samples by hydrodistillation for 3 h. The chemical analysis was performed by gas chromatography-mass spectrometry. Multivariate data analysis was employed to explain the interspecies relationships among Solidago spp. The results revealed the similarity among Solidago spp. EO profiles, which were dominated by monoterpenes and oxygenated compound fractions. Solidago spp. differed in species distinctive terpenes and their distribution between accessions and plant parts. Volatile compound patterns confirmed the origin of Solidago × niederederi between Solidago canadensis and Solidago virgaurea, with the higher contribution of alien species than native ones. Correct taxonomic identification of species is highly essential for the targeted collection of raw material from the wild for different applications. Solidago spp. can be considered to be underutilized sources of bioactive secondary metabolites.

The Effect of Traditional and Cyclodextrin-Assisted Extraction Methods on Trifolium pratense L. (Red Clover) Extracts Antioxidant Potential.

Red clover is the subject of numerous studies because of its antioxidant properties, the positive influence of isoflavones on the health, and its potential use in the prevention and treatment of chronic diseases. The right excipients, such as cyclodextrins, can increase the profile of valuable phenolic compounds in extraction media to obtain rich in antioxidants, extracts that can be used in nutraceuticals production. The aim of this study was to investigate and compare the total phenolic content, flavonoid content, and antioxidant activity of red clover aerial parts, aqueous and ethanolic extracts prepared using traditional and cyclodextrins-assisted methods. The antioxidant activity of the extracts was established using ABTS, DPPH, FRAP, and ABTS-post column methods. It was determined that cyclodextrins significantly increased total phenolic content (compared with control)-using ß-cyclodextrin 20.29% (in aqueous samples); γ-cyclodextrin 22.26% (in ethanolic samples). All the samples prepared with excipients demonstrated a strong relationship between total phenolic content and DPPH assay. Study showed that for extraction with water, the highest amounts of phenolic compounds, flavonoids and antioxidant activity will be achieved with ß-cyclodextrin, but extractions with ethanol will give the best results with γ-cyclodextrin. Therefore, cyclodextrins are a great and safe tool for obtaining rich, red clover flower extracts that are high in antioxidant activity, which can be used in the pharmaceutical industry for nutraceuticals production.

Effect of the Amount of Polysorbate 80 and Oregano Essential Oil on the Emulsion Stability and Characterization Properties of Sodium Alginate Microcapsules.

Essential oils have a high volatility that leads to evaporation and loss of their pharmacological effect when exposed to the environment. The objectives of the present work were to prepare microcapsules with oregano essential oil by extrusion using sodium alginate as a shell material and non-ionic surfactant polysorbate 80 as an emulsifier to stabilize the emulsion. The present study was aimed to evaluate the physical parameters of microcapsules and to compare the influence of the amount of emulsifier and the essential oil-to-emulsifier ratio on the capsules' physical parameters and encapsulation efficiency; to our knowledge, the existing research had not yet revealed whether unstable emulsion affects the encapsulation efficiency of oregano essential oil. This study showed that increasing the emulsifier amount in the formulation significantly influenced encapsulation efficiency and particle size. Moreover, increasing the emulsion stability positively influenced the encapsulation efficiency. The emulsion creaming index depended on the emulsifier amount in the formulation: the highest creaming index (%) was obtained with the highest amount of polysorbate 80. However, the essential oil-to-polysorbate 80 ratio and essential oil amount did not affect the hardness of the microcapsules (p > 0.05). In conclusion, the obtained results could be promising information for production of microcapsules. Despite the fact that microencapsulation of essential oils is a promising and extremely attractive application area for the pharmaceutical industry, further basic research needs to be carried out.

Bio-guided bioactive profiling and HPLC-DAD fingerprinting of Ukrainian saffron (Crocus sativus stigmas): moving from correlation toward causation.

BACKGROUND: Saffron or stigmas of Crocus sativus L. is one of the most valuable food products with interesting health-promoting properties. C. sativus has been widely used as a coloring and flavoring agent. Stigmas secondary metabolites showed potent cytotoxic effects in previous reports. METHODS: The present study investigated the chemical composition and the cytotoxic effect of Ukrainian saffron crude extracts and individual compounds against melanoma IGR39, triple-negative breast cancer MDA-MB-231, and glioblastoma U-87 cell lines in vitro using MTT assay. Several bioactivity in vitro assays were performed. The chemical profile of the water and hydroethanolic (70%, v/v) crude extracts of saffron stigmas was elucidated by HPLC-DAD analysis. RESULTS: Seven compounds were identified including crocin, picrocrocin, safranal, rutin, apigenin, caffeic acid, ferulic acid. Crocin, picrocrocin, safranal, rutin, and apigenin were the major active constituents of Ukrainian C. sativus stigmas. The hydroethanolic extract significantly reduced the viability of MDA-MB-231 and IGR39 cells and the effect was more potent in comparison with the water extract. However, the water extract was almost 5.6 times more active against the U-87 cell line (EC50 of the water extract against U-87 was 0.15 ± 0.02 mg/mL, and EC50 of the hydroethanolic extract was 0.83 ± 0.03 mg/mL). The pure compounds, apigenin, and caffeic acid also showed high cytotoxic activity against breast cancer, melanoma, and glioblastoma cell lines. The screening of the biological activities of stigmas water extract (up to 100 µg/mL) including anti-allergic, anti-virus, anti-neuraminidase, and anti-inflammatory effects revealed its inhibitory activity against neuraminidase enzyme by 41%. CONCLUSIONS: The presented results revealed the qualitative and quantitative chemical composition and biological activity of Crocus sativus stigmas from Ukraine as a source of natural anticancer and neuraminidase inhibitory agents. The results of the extracts' bioactivity suggested future potential applications of saffron as a natural remedy against several cancers.

Ophthalmic In Situ Gels with Balsam Poplar Buds Extract: Formulation, Rheological Characterization, and Quality Evaluation.

Balsam poplar buds are a raw material with a high content of polyphenols. Various polyphenols are known for their anti-inflammatory and antioxidant properties. In this study, an aqueous extract of balsam poplar buds was prepared in order to use environmentally friendly and non-aggressive solvents. The aqueous extract was lyophilized, and a 1% aqueous solution of lyophilized balsam poplar buds extract (L1) was prepared. L1 solution was used as a source of polyphenols for the production of ophthalmic in situ gels, so as to develop a product featuring antioxidant properties. Poloxamer 407 (P407) and hydroxypropyl methylcellulose (HPMC) were selected as gelling agents for the in situ gels. In order to select the formulations with the best conditions of use, formulations of different polymer concentrations (P407-10%, 12%, 15%; HPMC-0.5%, 0.75%) were prepared, choosing the same amount of the active polyphenol source L1. The physicochemical properties, rheological parameters, stability, and irritant effect on the rabbit corneal cell line (SIRC) were evaluated. Formulations in which P407 and HMPC concentrations were 10/0.75% and 12%/0.75% reached a gelation point close to the ocular surface temperature; the gels remained stable for 30 days and did not cause an irritant effect on the SIRC cell line.

Pharmacological Potential and Chemical Composition of Crocus sativus Leaf Extracts.

Crocus sativus L. (saffron) has been traditionally used as a food coloring or flavoring agent, but recent research has shown its potent pharmacological activity to tackle several health-related conditions. Crocus sp. leaves, and petals are the by-products of saffron production and are not usually used in the medicine or food industries. The present study was designed to determine the chemical composition of the water and ethanolic extracts of C. sativus leaves and test their cytotoxic activity against melanoma (IGR39) and triple-negative breast cancer (MDA-MB-231) cell lines by MTT assay. We also determined their anti-allergic, anti-inflammatory, and anti-viral activities. HPLC fingerprint analysis showed the presence of 16 compounds, including hydroxycinnamic acids, xanthones, flavonoids, and isoflavonoids, which could contribute to the extracts' biological activities. For the first time, compounds such as tectoridin, iristectorigenin B, nigricin, and irigenin were identified in Crocus leaf extracts. The results showed that mangiferin (up to 2 mg/g dry weight) and isoorientin (8.5 mg/g dry weight) were the major active ingredients in the leaf extracts. The ethanolic extract reduced the viability of IGR39 and MDA-MB-231 cancer cells with EC50 = 410 ± 100 and 330 ± 40 µg/mL, respectively. It was more active than the aqueous extract. Kaempferol and quercetin were identified as the most active compounds. Our results showed that Crocus leaves contain secondary metabolites with potent cytotoxic and antioxidant activities.

Qualitative and Quantitative Analysis of Ukrainian Iris Species: A Fresh Look on Their Antioxidant Content and Biological Activities.

The major groups of antioxidant compounds (isoflavonoids, xanthones, hydroxycinnamic acids) in the rhizome methanol extracts of four Ukrainian Iris sp. (Iris pallida, Iris hungarica, Iris sibirica, and Iris variegata) were qualitatively and quantitatively analyzed using HPLC-DAD and UPLC-MS/MS. Gallic acid, caffeic acid, mangiferin, tectoridin, irigenin, iristectorigenin B, irisolidone, 5,6-dihydroxy-7,8,3',5'-tetramethoxyisoflavone, irisolidone-7-O-ß-d-glucopyranoside, germanaism B, and nigricin were recognized by comparing their UV/MS spectra, chromatographic retention time (tR) with those of standard reference compounds. I. hungarica and I. variegata showed the highest total amount of phenolic compounds. Germanaism B was the most abundant component in the rhizomes of I. variegata (7.089 ± 0.032 mg/g) and I. hungarica (6.285 ± 0.030 mg/g). The compound analyses showed good calibration curve linearity (r2 > 0.999) and low detection and quantifications limit. These results validated the method for its use in the simultaneous quantitative evaluation of phenolic compounds in the studied Iris sp. I. hungarica and I. variegata rhizomes exhibited antioxidant activity, as demonstrated by the HPLC-ABTS system and NRF2 expression assay and anti-inflammatory activity on respiratory burst in human neutrophils. Moreover, the extracts showed anti-allergic and cytotoxic effects against cancer cells. Anti-coronavirus 229E and lipid formation activities were also evaluated. In summary, potent antioxidant marker compounds were identified in the examined Iris sp.

Biologically active compounds and pharmacological activities of species of the genus Crocus: A review.

The present article is the first comprehensive review on the chemical composition and pharmacological activities of the raw materials of Crocus species. In the present review, data on chemical constituents and pharmacological profile of Crocus sativus stigmas, as well as of other plant parts (perianth, stamens, leaves, corms) of different Crocus spp. are given. This review discusses all the classes of compounds (carotenoids, flavonoids, anthocyanins, terpenoids, phenol carboxylic acids, etc.) detected in raw materials of Crocus plants providing information on the current state of knowledge on phytochemicals of Crocus species. Almost all structural formulas of the compounds identified and isolated from Crocus species are given; all compounds are presented in accordance with the types of the studied raw materials. The latest hypotheses relating to the biosynthesis pathways of the main biologically active compounds of saffron (crocin, picrocrocin, safranal), as well as chemotaxonomy of Crocus genus are briefly summarized. The present review discusses the most thoroughly studied pharmacological activities (namely, antioxidant, antiparasitic, hypolipidemic, antihypertensive, immunomodulatory, antimicrobial, antitumor, cytotoxic, antidepressant) of saffron stigmas extracts, of its individual phytochemicals (safranal, crocin, crocetin), as well as pharmacological activities of raw materials of other Crocus species. This comprehensive review will be informative for scientists searching for new properties of saffron stigmas, as well as for saffron producers, since the present review highlights the prospects for the use of waste products in the production of the expensive spice. In addition, the present review provides information on pharmacological properties and composition of other Crocus species as promising medicinal and food plants. In the present review the emphasis will be put on the chemical constituents of Crocus species and the intraspecies variation in phytochemicals and pharmacological activities.

GC-MS Analysis of the Composition of the Extracts and Essential Oil from Myristica fragrans Seeds Using Magnesium Aluminometasilicate as Excipient.

Myristica fragrans (f. Myristicaceae) seeds are better known as a spice, but their chemical compounds may have a pharmacological effect. The yield of their composition of extracts and essential oils differs due to different methodologies. The aim of this study was to evaluate an excipient material-magnesium aluminometasilicate-and to determine its influence on the qualitative composition of nutmeg extracts and essential oils. Furthermore, we wanted to compare the yield of essential oil. The extracts were prepared by maceration (M) and ultrasound bath-assisted extraction (UAE), and the essential oil-by hydrodistillation (HD). Conventional methods (UAE, HD) were modified with magnesium aluminometasilicate. The samples were analyzed by gas chromatography-mass spectrometry (GC-MS) method. From 16 to 19 chemical compounds were obtained using UAE with magnesium aluminometasilicate, while only 8 to 13 compounds were obtained using UAE without an excipient. Using our conditions and plant material, for the first time eight new chemical compounds in nutmeg essential oil were identified. Two of these compounds (γ-amorphene and cis-α-bergamotene) were obtained with the use of excipient, the other six (ß-copaene, bergamotene, citronellyl decanoate, cubebol, cubenene, orthodene) by conventional hydrodistillation. Magnesium aluminometasilicate significantly increased the quantity of sabinene (from 6.53% to 61.42%) and limonene (from 0% to 5.62%) in essential oil. The yield of the essential oil from nutmeg seeds was significantly higher using magnesium aluminometasilicate; it increased from 5.25 ± 0.04% to 10.43 ± 0.09%.

Hilic MS/MS determination of amino acids in herbs of Fumaria schleicheri L., Ocimum basilicum L., and leaves of Corylus avellana L.

The aim of research was to study the content of amino acids using in extracts of Fumaria schleicheri L., Ocimum basilicum L., and Corylus avellana L. by HILIC MS/MS method. Separation of amino acids in the samples was carried out with Acquity H-class UPLC system (Waters, Milford, USA) equipped with SeQuant ZIC-Hilic collumn (2.1 × 150 mm, 3.5 µm) (Merck Millipore, Darmstadt, Germany). The MS/MS fragment ion chromatograms of the test solutions established the presence of 19 amino acids. The obtained results have shown that O. basilicum L. characterized the highest concentrations of different neurogenic amino acids (128.1 mg/kg), comparing with F. schleicheri L. and C. avellana L. (57.72 and 52.91 mg/kg, respectively).

Development of antioxidant food packaging materials containing eugenol for extending display life of fresh beef.

In this study, clove essential oil (CL) or eugenol (EU) containing cellulose acetate (CA) or acrylic component/hydrophobically modified starch (AC/S) coatings on corona treated oriented polypropylene film (OPP) were designed and investigated for their possible applications as antioxidant packaging materials for fresh meat. The antioxidant properties of the coatings were investigated by Vapour Phase-DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. The CA coatings containing CL or EU showed 43-92% and 43-94% inhibition against DPPH free radicals through the vapour phase, respectively, whereas AC/S/CL and AC/S/EU coatings resulted in DPPH inhibition of 21-65% and 25-84%, respectively. AC/S/EU and CA/EU coatings on OPP containing from 0.32 ±â€¯0.03 to 6.40 ±â€¯0.14 g/m2 of EU were used to prepare packaging for fresh beef (Longissimus thoracis). After 14 days, the lipid oxidation in beef steaks kept in control and antioxidant packages was 3.33 and 1.00-1.22 mg of malondialdehyde per kilogram of meat, respectively. Moreover, red colour of beef in antioxidant packages was retained.

Evaluation of phenolic antioxidant content in organically and conventionally grown buckwheat herb crop and its regrowth.

BACKGROUND: Buckwheat herb is known in the pharmaceutical industry as a material rich in phenolics. Buckwheat is also capable of regrowing and producing an additional harvest in the same year. Although buckwheat herb is a popular material, it is not known whether the regrowth has features of the same quality as the first harvest. Therefore, using the herb of 15 varieties of buckwheat cultivated in Lithuania, the present study aimed to examine biometric properties, phenolic content and antioxidant activity as material quality indicators for buckwheat herb and its regrowth under organic and conventional farming conditions. RESULTS: The highest amount of rutin was indicated in the organically cultivated regrowth of 2015. Buckwheat material accumulated significantly (P < 0.05) lower amounts of other phenolics. A significant correlation between phenolic content and antioxidant activity was observed in the herb and its regrowth in both farming systems. CONCLUSION: Regrowth was of similar quality to the first harvest in terms of phenolic content and antioxidant activity, although a downward trend in several biometric features was observed: the height of the regrowth plants was up to two-fold smaller and biomass was five- to nine-fold lower compared to that of the first harvest plants. Organic and industrial farming systems were determined to produce plants of similar quality. © 2016 Society of Chemical Industry.

Changes in the content of bioactive substances among Hypericum montbretii populations from Turkey

In the present study, we investigated the variation in the content of seventeen secondary metabolites among Hypericum montbretii Spach., Hypericaceae, populations from five different growing zones in Turkey for the first time.The plants were collected at full flowering, and after they were dried at room temperature, they were assayed for chemical contents by HPLC. Chemical constituents of plants varied significantly among populations except for 2,4-dihydroxybenzoic acid which was accumulated at similar levels. Plants from population - 1 yielded the highest amount of hypericin and pseudohypericin (1.27 and 2.97 mg/g, respectively) while hyperforin and adhyperforin accumulations were the highest in plants from population - 2 (6.64 and 1.24 mg/g, respectively). (+)-Catechin and (-)-epicatechin were accumulated at significantly higher levels by plants of population - 4 (1.54 and 4.35 mg/g, respectively). The highest accumulation level of the rest compounds namely, chlorogenic and neochlorogenic acids, amentoflavone, hyperoside, isoquercitrin, quercitrin, quercetin, avicularin and rutin was reached in plants from population-5 (2.64, 4.37, 2.35, 10.26, 3.52, 4.37, 1.55, 1.56 and 20.54 mg/g, respectively). The pronounced chemical diversity between populations is discussed to possibly be the result of different environmental, morphological and genetic factors.

Antioxidant effects of Camellia sinensis L. extract in patients with type 2 diabetes.

The prevalence of diabetes mellitus (DM) has dramatically increased in the past decade. Furthermore, increasing evidence from research shows that oxidative stress (OS) plays a crucial role in the pathogenesis of diabetes and in its complications. A search for ways to reduce oxidative damage has become the focus of interest for the majority of scientists. In this study, we determined the radical scavenging activity of single green tea constituents by using an on-line high performance liquid chromatography (HPLC)-2,2-diphenyl-1-picrylhydrazyl (DPPH) method and evaluated the antioxidant effects on type 2 diabetic patients by performing a double-blind, placebo-controlled study. Epigallocatechin gallate was identified as the most potent antioxidant, contributing approximately 50% of the total antioxidant capacity of green tea extract. We also found a statistically significant decrement of lipid peroxidation markers in patients treated with green tea extract after 9 months or after 18 months of follow-up. Overall, these findings are attractive for diabetic patients, helping them to keep a high level of performance and well-being, which ultimately may delay the time of disability and reduce mortality.

Chemical constituents of Hypericum adenotrichum Spach, an endemic Turkish species.

The present study was conducted out to determine hyperforin, hypericin, pseudohypericin, chlorogenic acid, rutin, hyperoside, quercitrin, quercetin, kaempferol, apigenin-7-O-glucoside and amentoflavone contents of Hypericum adenotrichum, an endemic plant species to Turkey. The aerial parts representing a total of 30 individuals were collected at full flowering, dried at room temperature and assayed for secondary metabolite concentrations by HPLC. All of the chemicals were detected at various levels except for hyperforin. This is the first report on polar chemistry of this endemic species.

HPLC with UV or mass spectrometric detection for quantifying endogenous uracil and dihydrouracil in human plasma.

BACKGROUND: We developed and compared 2 different methods for quantifying uracil (U) and dihydrouracil (UH(2)) in BSA and human plasma. Special attention was paid to the selectivity/specificity and the absence of a matrix effect. The UH(2)/U ratio is intended as a biomarker to identify patients with deficiency in 5-fluorouracil metabolism. METHODS: We quantified U and UH(2) with 2 liquid chromatography methods after solid-phase extraction, one with UV detection (LC-UV) and the other with mass spectrometric detection (LC-MS). We selected 2 internal standards to prevent the risk of interferences. Separation was achieved with a Waters Atlantis dC18 column (LC-MS) or a Waters SymmetryShield RP18 column connected with an Atlantis dC18 (LC-UV). Mass spectrometric data were acquired in single-ion monitoring mode. RESULTS: Assay imprecision in BSA solution was <15% (LC-UV) and <12% (LC-MS); in plasma, assay imprecision was <9.5% and <9.0%, respectively. Recoveries were 88.2%-110% (LC-UV) and 94.8%-107% (LC-MS). Extraction efficiencies were >or=89.0%. In BSA, the lower limits of quantification for U and UH(2) were 2.5 microg/L and 6.25 microg/L, respectively, for the LC-UV method and 2.5 microg/L and 3.1 microg/L for LC-MS. The corresponding values in plasma were 11.6 microg/L and 21.5 microg/L, and 4.1 microg/L and 12.1 microg/L. CONCLUSIONS: To estimate endogenous U and UH(2) concentrations and their ratio, we recommend the use of a drug-free human plasma pool in which baseline U and UH(2) concentrations have previously been measured with the standard-addition method. Our LC-MS method, which has the better test performance and is useful for measuring UH(2)/U ratios in cancer patients, is preferred when this equipment is available.