Phytochemical Profile and Biological Activity of the Ethanolic Extract from the Aerial Part of Crocus alatavicus Regel & Semen Growing Wildly in Southern Kazakhstan.

The composition of the ethanolic extract from the aerial parts of Crocus alatavicus Regel & Semen from southern Kazakhstan spontaneous flora was analyzed together with the determination of its antibacterial, antifungal, antiviral and anticancer activity. The phytochemical profile analysis by high-performance liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry (HPLC/ESI-QTOF-MS) revealed the presence of multiple kaempferol derivatives. High-performance reverse-phase liquid chromatography combined with a photodiode-array detection (RP-HPLC/PDA) found that kaempferol 3-O-dihexoside and kaempferol 3-O-acyltetrahexoside accounted for 70.5% of the kaempferol derivatives. The minimum inhibitory concentration (MIC) values of the extract for all the tested reference microorganisms were high, reaching 10 mg/mL for yeasts and 20 mg/mL for bacteria. In contrast, antiviral activity was observed at 2 mg/mL, resulting in the inhibition of the HSV-1-induced cytopathic effect and the reduction in virus infectious titer by 1.96 log, as well as the viral load by 0.85 log. Among the tested prostate cancer cell lines, significant cytotoxic activity of the extract was noted only on the LNCaP cell line, with an IC50 value of 1.95 mg/mL. The LNCaP cell line treated with 2 mg/mL of the extract showed a noticeably reduced number of spindle-shaped cells with longer cellular projections, a significant increase in the peak corresponding to the population of apoptotic cells in the sub-G1 phase and a decreased intracellular glutathione (GSH) level, suggesting the prooxidative properties of the extract. The obtained data provide novel information about the flavonoids present in the aerial part of C. alatavicus and suggest its potential application as a source of the compounds active against HSV-1 and metastatic, androgen-sensitive prostate cancer.

Clove Essential Oil and Its Main Constituent, Eugenol, as Potential Natural Antifungals against Candida spp. Alone or in Combination with Other Antimycotics Due to Synergistic Interactions.

The occurrence of candidiasis, including superficial infections, has recently increased dramatically, especially in immunocompromised patients. Their treatment is often ineffective due to the resistance of yeasts to antimycotics. Therefore, there is a need to search for new antifungals. The aim of this study was to determine the antifungal effect of clove essential oil (CEO) and eugenol (EUG) towards both reference and clinical Candida spp. strains isolated from the oral cavity of patients with hematological malignancies, and to investigate their mode of action and the interactions in combination with the selected antimycotics. These studies were performed using the broth microdilution method, tests with sorbitol and ergosterol, and a checkerboard technique, respectively. The CEO and EUG showed activity against all Candida strains with a minimal inhibitory concentration (MIC) in the range of 0.25-2 mg/mL. It was also found that both natural products bind to ergosterol in the yeast cell membrane. Moreover, the interactions between CEO and EUG with several antimycotics-cetylpyridinium chloride, chlorhexidine, silver nitrate and triclosan-showed synergistic or additive effects in combination, except nystatin. This study confirms that the studied compounds appear to be a very promising group of phytopharmaceuticals used topically in the treatment of superficial candidiasis. However, this requires further studies in vivo.

UPLC-MS-ESI-QTOF Analysis and Antifungal Activity of Aqueous Extracts of Spondias tuberosa.

This study aimed to identify the chemical composition of the Spondias tuberosa aqueous leaf and root extracts (EALST and EARST) and to evaluate their effect, comparatively, against opportunistic pathogenic fungi. Ultra-Performance Liquid Chromatography Coupled to a Quadrupole/Time of Flight System (UPLC-MS-ESI-QTOF) was employed for chemical analysis. Candida albicans and C. tropicalis standard strains and clinical isolates were used (CA INCQS 40006, CT INCQS 40042, CA URM 5974, and CT URM 4262). The 50% Inhibitory Concentration for the fungal population (IC50) was determined for both the intrinsic action of the extracts and the extract/fluconazole (FCZ) associations. The determination of the Minimum Fungicidal Concentration (MFC) and the verification of effects over fungal morphological transitions were performed by subculture in Petri dishes and humid chambers, respectively, both based on micro-dilution. UPLC-MS-ESI-QTOF analysis revealed the presence of phenolic and flavonoid compounds. The association of the extracts with fluconazole, resulted in IC50 values from 2.62 µg/mL to 308.96 µg/mL. The MFC of the extracts was ≥16,384 µg/mL for all tested strains, while fluconazole obtained an MFC of 8192 µg/mL against C. albicans strains. A reduction in MFC against CA URM 5974 (EALST: 2048 µg/mL and EARST: 1024 µg/mL) occurred in the extract/fluconazole association.

Analytical Assessment of Bioelements in Various Types of Black Teas from Different Geographical Origins in View of Chemometric Approach.

A comprehensive approach to the mineral composition of black teas of different origins was studied using the Flame Atomic Absorption Spectrometry (FAAS) method, supported by chemometric tools including Principal Component Analysis PCA) and Classification and Regression Trees (CART). Significant differences between the teas from seven countries (Japan, Nepal, Kenya, Iran, Sri Lanka, India, and China) were shown. K was the main element determined in all teas, with an average concentration of 11,649 mg/kg, followed by Ca, Mg and Mn. In general, regarding all investigated black teas, the element content was ranked in the following order: K > Ca > Mg > Mn > Fe > Na > Zn > Cu. The applied chemometric methods allowed us to recognize black tea clusters based on their mineral composition and place of cultivation, and allowed us to find correlations between particular elements in black teas. The performed analyses revealed interesting correlations between the concentration of various elements in black teas: K was negatively correlated with Na, Fe, Mn and Cu; K was positively correlated with the content of Ca and Mg. Significant positive correlations between Mn and Fe and Mn and Zn in the studied black tea samples were also revealed. It was shown that mineral composition may be a significant factor regarding the origin of the black tea, not only considering the country, but also the region or province.

Phytochemical Fingerprinting and In Vitro Antimicrobial and Antioxidant Activity of the Aerial Parts of Thymus marschallianus Willd. and Thymus seravschanicus Klokov Growing Widely in Southern Kazakhstan.

The chemical composition of the hydroethanolic extracts (60% v/v) from the aerial parts of Thymus marschallianus Willd (TM) and Thymus seravschanicus Klokov (TS) from Southern Kazakhstan flora was analyzed together with their hexane fractions. Determination of antibacterial, antifungal and antioxidant activities of both extracts was also performed. RP-HPLC/PDA and HPLC/ESI-QTOF-MS showed that there were some differences between the composition of both extracts. The most characteristic components of TM were rosmarinic acid, protocatechuic acid, luteolin 7-O-glucoside, and apigenin 7-O-glucuronide, while protocatechuic acid, luteolin 7-O-glucoside, luteolin 7-O-glucuronide, and eriodictyol predominated in TS. The content of polyplenols was higher in TS than in TM. The GC-MS analysis of the volatile fraction of both examined extracts revealed the presence of thymol and carvacrol. Additionally, sesquiterpenoids, fatty acids, and their ethyl esters were found in TM, and fatty acid methyl esters in TS. The antioxidant activity of both extracts was similar. The antibacterial activity of TS extract was somewhat higher than TM, while antifungal activity was the same. TS extract was the most active against Helicobacter pylori ATCC 43504 with MIC (minimal inhibitory concentration) = 0.625 mg/mL, exerting a bactericidal effect. The obtained data provide novel information about the phytochemistry of both thyme species and suggest new potential application of TS as a source of bioactive compounds, especially with anti-H. pylori activity.

Production of enriched in B vitamins biomass of Yarrowia lipolytica grown in biofuel waste.

Yarrowia lipolytica as an oleaginous yeast is capable of growing in various non-conventional hydrophobic substrate types, especially industrial wastes. In this study, the content of thiamine (vitamin B1), riboflavin (vitamin B2), pyridoxine (vitamin B6), biotin (vitamin B7) and folic acid (vitamin B9) in the wet biomass of Y. lipolytica strains cultivated in biofuel waste (SK medium), compared to the standard laboratory YPD medium, was assessed. Additionally, the biomass of Y. lipolytica A-101 grown in biofuel waste (SK medium) was dried and examined for B vitamins concentration according to the recommended microbial methods by AOAC Official Methods. The mean values of these vitamins per 100 g of dry weight of Y. lipolytica grown in biofuel waste (SK medium) were as follows: thiamine 1.3 mg/100 g, riboflavin 5.3 mg/100 g, pyridoxine 4.9 mg/100 g, biotin 20.0 µg/100 g, and folic acid 249 µg/100 g. We have demonstrated that the dried biomass is a good source of B vitamins which can be used as nutraceuticals to supplement human diet, especially for people at risk of B vitamin deficiencies in developed countries. Moreover, the biodegradation of biofuel waste by Y. lipolytica is desired for environmental protection.

Chemical and Nutritional Compounds of Different Parts of Lemongrass (Cymbopogon citratus (DC) Stapf.) Cultivated in Temperate Climate of Poland.

Lemongrass (Cymbopogon citratus (DC) Stapf.) is a perennial plant indigenous to semi-tropical regions of Asia and cultivated in other semi-tropical countries. The present study aimed to examine the key chemical constituents of various parts of lemongrass cultivated in the temperate climate of Poland. The content of essential oil and its composition were determined in 4 plant parts: leaves (part C), overground shoots (part B), underground shoots (part A), and roots (part R). Moreover, the content of dry weight, chlorophyll, polyphenols and macro- and microelements was determined in the edible parts (excluding roots). The essential oil from the aerial part predominantly contained neral (> 30%) and geranial (> 40%), which is consistent with the data reported in literature; the main component of essential oil (EO) from the underground part was elemol (65%); interestingly, such a high concentration of it was found for the first time. The concentration of chlorophyll was found to be higher in leaves, as compared to parts B and A. The highest level of potassium, magnesium, zinc and sodium was found in part A while of calcium and copper in leaves. The quality of lemongrass raw materials grown in temperate climates did not differ significantly from those obtained in warmer regions. The study findings confirmed the usefulness of leaves as a raw material for the preparation of infusions (higher concentration of pigments, polyphenols and EO) and of near-ground parts of a plant as a culinary material (a higher content of macroelements at lower concentrations of green pigments and dry weight).

The In Vitro Activity of Essential Oils against Helicobacter Pylori Growth and Urease Activity.

The anti-H. pylori properties of 26 different commercial essential oils were examined in vitro by MIC (minimal inhibitory concentration) determination for the reference strain H. pylori ATCC 43504. We selected 9 essential oils with different anti-Helicobacter activities and established their phytochemical composition and urease inhibition activities. Phytochemical analysis of the selected essential oils by GC-MS method and antioxidant activity were performed. The phenol red method was used to screen the effect of essential oils on urease activity expressed as IC50 (the half of maximal inhibitory concentration). The most active essential oils, with MIC = 15.6 mg/L, were thyme, lemongrass, cedarwood and lemon balm oils; MIC = 31.3 mg/L-oregano oil; MIC = 62.5 mg/L-tea tree oil; MIC = 125 mg/L-pine needle, lemon and silver fir oils with bactericidal effect. Urease activity was inhibited by these oils with IC50 ranged from 5.3 to > 1049.9 mg/L. The most active was cedarwood oil (IC50 = 5.3 mg/L), inhibiting urease at sub-MIC concentrations (MIC = 15.6 mg/L). The statistical principal component analysis allowed for the division of the oils into three phytochemical groups differing in their anti-H. pylori activity. To summarize, the activity in vitro of the five essential oils silver fir, pine needle, tea tree, lemongrass, and cedarwood oils against H. pylori was found in this paper for the first time. The most active against clinical strains of H. pylori were cedar wood and oregano oils. Moreover, cedarwood oil inhibited the urease activity at subinhibitory concentrations. This essential oil can be regarded as a useful component of the plant preparations supporting the eradication H. pylori therapy.

Phytochemical composition of wormwood (Artemisia gmelinii) extracts in respect of their antimicrobial activity.

BACKGROUND: Extracts from medicinal plants with phytochemicals with known antimicrobial properties can be an effective adjunct in the complex treatment of infectious diseases. This study aimed to evaluate the antimicrobial activity of wormwood extracts collected in Kazakhstan (Artemisia gmelinii Weber ex Stechm.), along with their phytochemical analysis. METHODS: The ethanolic and chloroform extracts were subjected to HPLC combined with quadrupole time-of-flight mass spectrometry method. For quantitative assessment of antimicrobial activity, minimal inhibitory concentration (MIC) of the tested extracts was determined by micro-dilution broth method for the panel of the reference microorganisms. Minimal bactericidal concentration (MBC) or minimal fungicidal concentration (MFC) were also determined. RESULTS: LC/MS analysis showed the presence of 13 compounds in the tested extracts, including flavonoids: apigenin, luteolin, rutin, two O-methylated flavonols (isorhamnetin, rhamnazine), coumarin compounds (umbelliferone, scopoletin and scopolin (scopoletin 7-glucoside), 3-hydroxycoumarin and 4-hydroxycoumarin), chlorogenic acid and two dicaffeoylquinic acid isomers. Quantitative HPLC analysis showed that umbelliferone was dominant in the chloroform extract while chlorogenic acid was identified as a main compound in the ethanolic extract. The antibacterial and antifungal activity of chloroform and ethanolic extracts was comparable. The most sensitive were the Gram-positive bacteria represented by staphylococci, Micrococcus luteus and Bacillus spp. (MIC = 1.25-5 mg/ml) and yeasts represented by Candida spp. (MIC = 2.5-5 mg/ml), irrespective of the assayed extract. CONCLUSIONS: Extracts of wormwood Artemisia gmelinii have shown a wide spectrum of antibacterial and antifungal activity. Luteolin, rutin, isorhamnetin and scopolin were identified in A. gmelinii species for the first time. The determining of the most potential compounds of Artemisia gmelinii can be used to develop effective antibacterial and antifungal agents.

LC-QTOF-MS Analysis and Activity Profiles of Popular Antioxidant Dietary Supplements in Terms of Quality Control.

The dietary supplements with claimed antioxidant activity constitute a substantial part of the dietary supplement market. In this study, we performed the LC-QTOF-MS analysis and investigated the activity profiles of popular antioxidant dietary supplements from different chemical groups in terms of quality control. The commonly used antioxidant tests and statistical analysis revealed that substantial part of the results was comparable if 1 g sample was considered, but while comparing single and daily doses, significant differences in antioxidant values were noticed in all assays. The best antioxidant activity was obtained in ORAC assay (from 142 to 13814 µM of Trolox equivalents per 1 g of sample), and the strongest correlation occurred between TPC and ORAC. The LC-QTOF-MS analysis revealed that catechins were present in samples having the best antioxidant activity and that dietary supplements showing the weakest activity contained very small amount of any chemical constituents.

Estimation of Potential Availability of Essential Oil in Some Brands of Herbal Teas and Herbal Dietary Supplements.

INTRODUCTION: The aim of the study was to estimate potential availability of essential oil in some brands of herbal products. METHODS: A comparison was performed on the basis of the essential oil yield in the unprocessed raw materials such as leaves of peppermint and lemon balm and inflorescence of chamomile as well as herbal tea bags and in dietary supplements. The yield of essential oil was determined by distillation. Essential oil was analyzed by GC-FID and GC-MS. RESULTS: It was found that the average potential availability of essential oils in the products such as dietary supplements for the doses recommended by the producers is lower than in the corresponding tea infusions: for peppermint formulations approximately 6-fold lower, for the formulations with lemon balm about 4-fold lower, and for the chamomile preparations about 3-fold lower. It was found that essential oils extracted from herbal teas have a similar chemical profile with characteristic deviations in the amount of individual components, which arise from the origin of the raw material. DISCUSSION: In contrast to homogenous pharmaceutical herbal mixtures consistent with, the Pharmacopoeia requirements, herbal teas (available in grocery stores) and dietary supplements are often out of control in terms of the yield and composition of the essential oil, which is primarily responsible for the health benefits and aromatic qualities of these products. Analysis of the composition of the dietary supplements showed that they contain on average significantly lower amounts of plant material compared to the herbal teas.

GC-MS analysis of essential oils from Salvia officinalis L.: comparison of extraction methods of the volatile components.

In this paper, comparison of the volatile components composition in the samples obtained by hydrodistillation and solid-phase microextraction of Salvia officinalis was described. Different sample preparation techniques showed considerable differences in volatiles composition, especially with respect to sesqui- and diterpenoids. The comparison of the sage essential oil obtained by hydrodistillation in the Deryng and Clevenger type apparatus, according to the pharmacopoeial methods (FP VI and VII), showed the presence of the same terpenoids in both essential oils, however, the relative percentage composition of the components were different. These differences are caused by the different extraction times used in both methods. Since each essential oil to be admitted to medicinal use should meet requirements regarding the composition of major chemical components, the minimum time for the hydrodistillation of the essential oils from sage should be 1 h.

Phenolic acids content, antioxidant and antimicrobial activity of Ligusticum mutellina L.

A simple HPLC method has been used for separation and quantitative analysis of the phenolic acids in the methanolic extracts of Ligusticum mutellina aerial parts. Chlorogenic acid was the predominant phenolic acid. Additionally, gallic, p-OH-benzoic, caffeic, p-coumaric and ferulic acids were identified. Moderate antibacterial and antifungal activity (MIC = 1.25-2.5 mg mL(-1)) was observed for the methanol extract of L. mutellina herb received from plants in flowering stage against a broad spectrum of bacteria. Micrococcus luteus, Pseudomonas aeruginosa and Candida spp. were the most sensitive to this plant material. Total phenolic content for the methanol extract of L. mutellina herb received from plants in flowering stage was 1.56 g of chlorogenic acid equivalents/100 g dry weight. The methanol extract of L. mutellina herb received from plants in flowering stage showed antioxidant activity with DPPH (IC50 value of 0.40 mg mL(-1)) and with ABTS (IC50 value of 8.65 mg mL(-1)).

Cytotoxicity, antioxidant activity and an effect on CYP3A4 and CYP2D6 of Mutellina purpurea L. extracts.

Mutellina purpurea is an aromatic Apiaceae plant known as Alpine lovage. Its polar extracts consist of phenolic acids, tannins and flavonoids. The cytotoxic effect of methanolic and aqueous extracts from M. purpurea was studied on the most frequently used cell lines: HeLa and BHK-21. Taking into account that the natural products are often used with other medicines there is a risk of reciprocal interaction on the metabolic level. Thus, the influence of M. purpurea extracts was investigated on the activity of CYP2D6 and CYP3A4, which are the most important P450 isoenzymes from the pharmacological and toxicological points of view. Additionally, because M. purpurea contains phenolic compounds, the antioxidative properties of this plant extracts were also studied and compared.