Trehalose determination in Norway spruce (Picea abies) roots. Analytics matters.

We present concise results of method validation for trehalose quantitation by LC-MS/MS in spruce ectomycorrhizal roots in order to describe spruce health status, mainly in connection to contamination by a pathogenic fungus, Gemmamyces piceae. The procedure is based on Rogatsky et al. (2005) developed for human plasma. We found out that the best extraction yield was achieved with 80% methanol/water (v/v) solution and optimal extraction temperature was set between 50-60°C. In contrast to previous papers, we minimized the activity of trehalase enzyme by putting root samples into liquid N2 immediately after root excavation, followed by freeze-drying in order to stop trehalase activity. Higher content of trehalose was recorded in healthy trees, confirming the idea that ectomycorrhiza plays a significant role in plant-pathogen interactions.

A comparative study of the HPLC-MS profiles and biological efficiency of different solvent leaf extracts of two African plants: Bersama abyssinica and Scoparia dulcis.

In the present study, two medicinal plants from Africa, namely Bersama abyssinica Fresen. and Scoparia dulcis L., were extracted using ethyl acetate, methanol, and water. The antioxidant, enzyme (α-amylase, α-glucosidase, acetyl- and butyrylcholinesterase, lipase, and tyrosinase) inhibitory action, and phytochemical profiles of extracts of Bersama abyssinica and Scoparia dulcis were determined. The aqueous (180.62 and 61.81 mg gallic acid equivalent/g extract, for B. abyssinica and S. dulcis respectively) and methanol (75.21 and 57.81 mg rutin equivalent/g extract, for B. abyssinica and S. dulcis, respectively) extracts contained high concentrations of phenolic and flavonoids, respectively. The ethyl acetate extracts of both plants were potent inhibitors of α-glucosidase and tyrosinase. Several phytochemical groups were determined by HPLC-MS/MS. The study tend to suggest that B. abyssinica and S. dulcis are potential candidates for the development of novel therapeutical agents.

Chemical Constituents and Biologic Activities of Sage Species: A Comparison between Salvia officinalis L., S. glutinosa L. and S. transsylvanica (Schur ex Griseb. & Schenk) Schur.

Even though Salvia genus is one of the most known and studied taxa of Lamiaceae family, the knowledge regarding the chemical composition and health-related benefits of some locally used Salvia species (mostly endemic) is still scarce. In this regard, the present work aims to evaluate the chemical profile and potential bioactivities of 70% (v/v) ethanolic extracts obtained from the less-studied S. transsylvanica and S. glutinosa in comparison with S. officinalis. HPLC-PDA analysis revealed the presence of rutin and catechin as the main compounds in the extracts of the three studied species (using the employed HPLC method), whereas the presence of naringenin was highlighted only in S. glutinosa extract. Chlorogenic acid, rutin and quercetin were identified and quantified for the first time in S. transsylvanica extracts. The in vitro antioxidant capacity of each extract was tested through complementary methods (phosphomolybdenum assay, DPPH, ABTS, CUPRAC and FRAP assays), and correlated with the presence of phenolics (especially flavonoids) in high amounts. The neuroprotective and antidiabetic abilities of S. officinalis (the most active as AChE, BChE and α-glucosidase inhibitor), S. glutinosa (the most active as α-amylase inhibitor) and S. transsylvanica were also studied. For each extract it was determined the antimicrobial, antifungal and cytotoxic effects using in vitro assays. The obtained results confirm the potential of S. transsylvanica and S. glutinosa as promising sources of bioactive compounds and as a starting point for further analyses.

Application of liquid-phase microextraction to the analysis of plant and herbal samples.

INTRODUCTION: The analysis of plant and herbal samples is a challenging task for analytical chemists due to the complexity of the matrix combined with the low concentration of analytes. In recent years different liquid-phase microextraction (LPME) techniques coupled with a variety of analytical equipment have been developed for the determination of both organic and inorganic analytes. OBJECTIVE: Over the past few years, the number of research papers in this field has shown a markedly growing tendency. Therefore, the purpose of this review paper is to summarise and critically evaluate research articles focused on the application of LPME techniques for the analysis of plant and herbal samples. RESULTS: Due to the complex nature of the samples, the direct application of LPME techniques to the analysis of plants has not often been done. LPME techniques as well as their modalities have been commonly applied in combination with other pretreatment techniques, including a solid-liquid extraction technique supported by mechanical agitation or auxiliary energies for plant analysis. Applications and the most important parameters are summarised in the tables. CONCLUSION: This review summarises the application of the LPME procedure and shows the major benefits of LPME, such as the low volume of solvents used, high enrichment factor, simplicity of operation and wide selection of applicable detection techniques. We can expect further development of microextraction analytical methods that focus on direct sample analysis with the application of green extraction solvents while fully automating procedures for the analysis of plant materials.

In Vitro Enzyme Inhibitory Properties, Secondary Metabolite Profiles and Multivariate Analysis of Five Seaweeds.

Seaweeds have been exploited as both food products and therapeutics to manage human ailments for centuries. This study investigated the metabolite profile of five seaweeds (Halimeda spp., Spyridia hypnoides (Bory de Saint-Vincent) Papenfuss, Valoniopsis pachynema (G. Martens) Børgesen, Gracilaria fergusonii J. Agardh and Amphiroa anceps (Lamarck) Decaisne using ultra-high-performance liquid chromatography coupled with electrospray ionization mass spectrometry (UHPLC-ESI-MS/MS). Furthermore, these seaweeds were assessed for antioxidant and inhibitory effects against α-amylase, α-glucosidase, acetyl-cholinesterase (AChE), butyryl-cholinesterase (BChE) and tyrosinase. Valoniopsis pachynema and A. anceps yielded the highest flavonoid (4.30 ± 0.29 mg RE/g) and phenolic content (7.83 ± 0.08 mg RE/g), respectively. Additionally, A. anceps exhibited significant antioxidant properties with all assays and significantly depressed BChE (IC50 = 6.68 ± 0.83 mg/mL) and α-amylase activities (IC50 = 5.34 ± 0.14 mg/mL). Interestingly, the five seaweeds revealed potent inhibitory effects against tyrosinase activity. In conclusion, A. anceps might be considered as a key source of phytoantioxidants and a potential candidate to develop nutritional supplements. Besides, the five tested seaweeds warrant further study and may be exploited as promising natural sources for managing hyperpigmentation.

Comprehensive approaches on the chemical constituents and pharmacological properties of flowers and leaves of American basil (Ocimum americanum L).

Ocimum americanum L. (Lamiaceae) is a common food condiment and also used in traditional medicine in the management of several human diseases. Nonetheless, there has been no effort to delineate the biological and phytochemical profiles of leaves and flowers prepared by different extractive solvents (ethyl acetate, methanol (MeOH), and water). The pharmacological potential of O. americanum extracts on pro-oxidant/pro-inflammatory mediators in rat colon specimens treated with lipopolysaccharide was investigated. In parallel, the inhibitory effects of the extracts on fungal and bacterial strains involved in ulcerative colitis were studied. Qualitative phytochemical analysis showed the presence of phenols, flavonoids, and tannins. Water extracts of flowers and leaves showed strong reducing and radicals scavenging potential. Both MeOH and ethyl acetate extracts of the leaves and flowers were able to inhibit acetylcholinesterase, butyrylcholinesterase, and tyrosinase. All the extracts inhibited the selected bacterial and fungal strains, while only ethyl acetate flower extract displayed antioxidant/anti-inflammatory effects in rat colon. The water and MeOH extracts stimulated colon lactate dehydrogenase (LDH) and serotonin (5-HT) and induced spontaneous migration of HCT116 cells. Future investigations should focus on the biological activity of isolated phytochemicals from the leaves and flowers of O. americanum, in order to clarify the mechanism(s) of action substantiating the observed pharmacological properties.

Qualitative Chemical Characterization and Multidirectional Biological Investigation of Leaves and Bark Extracts of Anogeissus leiocarpus (DC.) Guill. & Perr. (Combretaceae).

Anogeissus leiocarpus (DC.) Guill. & Perr. (Combretaceae) has a long history of use by folk populations for the management of multiple human ailments. Based on the published literature, there has been no attempt to conduct a comparative assessment of the biological activity and the phytochemical profiles of the leaves and stem bark of A. leiocarpus extracted using methanol, ethyl acetate, and water. By high-performance liquid chromatography with electrospray ionization mass spectrometric detection (HPLC-ESI-MSn) analysis, quinic, shikimic, gallic, and protocatechuic acids were tentatively identified from all the extracts, while chlorogenic, caffeic, ferulic, and dodecanedioic acids were only characterised from the leaves extracts. Additionally, a pharmacological study was carried out to evaluate potential protective effects that are induced by the extracts in rat colon and colon cancer HCT116 cell line. In general, the methanol and water extracts of A. leiocarpus leaves and stem bark showed potent radical scavenging and reducing properties. It was noted that the stem bark extracts were more potent antioxidants as compared to the leaves extracts. The methanol extract of A. leiocarpus leaves showed the highest acetyl (4.68 mg galantamine equivalent/g) and butyryl (4.0 mg galantamine equivalent/g) cholinesterase inhibition. Among ethyl acetate extracts, the pharmacological investigation suggested stem bark ethyl acetate extracts to be the most promising. This extract revealed ability to protect rat colon from lipopolysaccharide-induced oxidative stress, without exerting promoting effects on HCT116 cell line viability and migration. As a conclusion, A. leiocarpus represents a potential source of bioactive compounds in the development of novel therapeutic agents.

New insights into the chemical profiling, cytotoxicity and bioactivity of four Bunium species.

Bunium species have been reported to be used both as food and in traditional medicines. The scientific community has attempted to probe into the pharmacological and chemical profiles of this genus. Nonetheless, many species have not been investigated fully to date. In this study, we determined the phenolic components, antimicrobial, antioxidant, and enzyme inhibitory activities of aerial parts of four Bunium species (B. sayai, B. pinnatifolium, B. brachyactis and B. macrocarpum). Results showed that B. microcarpum and B. pinnatifolium were strong antioxidants as evidenced in the DPPH, ABTS, CUPRAC, and FRAP assays. B. brachyactis was the most effective metal chelator, and displayed high enzyme inhibition against cholinesterase, tyrosinase, amylase, glucosidase, and lipase. The four species showed varied antimicrobial activity against each microorganism. Overall, they showed high activity against P. mirabilis and E. coli (MIC and MBC <1 mg mL-1). B. brachyactis was more effective against Aspergillus versicolor compared to the standard drug ketoconazole. B. brachyactis was also more effective than both ketoconazole and bifonazole against Trichoderma viride. B. sayai was more effective than ketoconazole in inhibiting A. fumigatus. B. sayai was most non-toxic to HEK 293 (cellular viability = 117%) and HepG2 (cellular viability = 104%). The highest level of TPC was observed in B. pinnatifolium (35.94 mg GAE g-1) while B. microcarpum possessed the highest TFC (39.21 mg RE g-1). Seventy four compounds were detected in B. microcarpum, 70 in B. brachyactis, 66 in B. sayai, and 51 in B. pinnatifolium. Quinic acid, chlorogenic acid, pantothenic acid, esculin, isoquercitrin, rutin, apigenin, and scopoletin were present in all the four species. This study showed that the four Bunium species are good sources of biologically active compounds with pharmaceutical and nutraceutical potential.

Investigation of chemical profile, biological properties of Lotus corniculatus L. extracts and their apoptotic-autophagic effects on breast cancer cells.

This study aimed to reveal chemical profiles and biological activities of ethyl acetate (EA), methanol (MeOH), and water extracts of Lotus corniculatus. Ethnobotanical reports have indicated the importance of phytochemical properties of the genus Lotus. In this study, the effects of medicinal plant extracts on antioxidant (DPPH, ABTS, CUPRAC, FRAP, phosphomolybdenum, and metal chelating assays), enzyme inhibitory (on cholinesterase, tyrosinase, a-amylase and a-glucosidase), DNA protection and anticancer properties (including anti-proliferative, cell death and telomerase activity marker gene analysis, apoptotic DNA fragmentation analysis, cell migration test) were evaluated. According to chemical analysis, quercetin derivatives geraldol, isorhamnetin and kaempferol-O-coumaroylhexoside-O-deoxyhexoside isomers were dominant in the extracts. MeOH extracts showed the highest total flavonoids capacity with 21.13 mg RE/g. EA extract showed the strongest anti-amylase activity among the tested extracts. Water extract had the most protective activity against plasmid DNA. To indicate cell survival, MTT test was performed against human MCF-7 and MDA-MB-231 breast cancer cells. Half-maximal inhibitory concentration for cells were calculated and used for detection of mechanisms behind the cancer cell death. EA extract showed up-regulation of Bax proapoptotic gene and apoptotic DNA fragmentation activity on highly invasive MDA-MB-231 cells. Beclin-1 and LC3-II autophagy genes were higly expressed after treatment of MCF-7 cells with EA extracts. EA and MeOH extracts inhibited cell migration ability of both cancer cells. Linoleamide, was dominant component in EA extract and caused apoptosis on MDA-MB-231 breast cancer cells via increasing intranuclear Ca²+. The detailed mechanism behind the anticancer properties should be further investigated.

Chemical fingerprints, antioxidant, enzyme inhibitory, and cell assays of three extracts obtained from Sideritis ozturkii Aytaç & Aksoy: An endemic plant from Turkey.

This study was geared towards assessing the possible antioxidant, enzyme inhibitory, and cytotoxic activities of ethyl acetate, methanol, and water extracts of Sideritis ozturkii Aytaç & Aksoy. The phytochemical profiles of the studied extracts were characterised by HPLC-MS/MS. The methanol extract, rich in phenolics (78.04 mg gallic acid equivalent/g), exhibited the strongest antioxidant activities. However, the ethyl acetate extract was the most active extract in the enzyme inhibitory assays. The water extract of S. ozturkii (1 mg/ml, 48 h incubation) slightly inhibited (22%) growth of human breast cancer cell line (MDA-MB-231 cells). On the other hand, the ethyl acetate and methanol extracts showed strong inhibition (98% and 97%, respectively) of MDA-MB-231 cells and caused apoptotic cell death. Scientific data generated from this study further appraises the multiple biological activities of plants belonging to the Sideritis genus. In addition, preliminary evidence gathered from the current investigation advocates for further studies geared towards the preparation of therapeutic formulations from S. ozturkii.

Liquid Phase and Microwave-Assisted Extractions for Multicomponent Phenolic Pattern Determination of Five Romanian Galium Species Coupled with Bioassays.

Background: Galium is a plant rich in iridoid glycosides, flavonoids, anthraquinones, and small amounts of essential oils and vitamin C. Recent works showed the antibacterial, antifungal, antiparasitic, and antioxidant activity of this plant genus. Methods: For the determination of the multicomponent phenolic pattern, liquid phase microextraction procedures were applied, combined with HPLC-PDA instrument configuration in five Galium species aerial parts (G. verum, G. album, G. rivale, G. pseudoaristatum, and G. purpureum). Dispersive Liquid⁻Liquid MicroExtraction (DLLME) with NaCl and NAtural Deep Eutectic Solvent (NADES) medium and Ultrasound-Assisted (UA)-DLLME with ß-cyclodextrin medium were optimized. Results: The optimal DLLME conditions were found to be: 10 mg of the sample, 10% NaCl, 15% NADES or 1% ß-cyclodextrin as extraction solvent-400 µL of ethyl acetate as dispersive solvent-300 µL of ethanol, vortex time-30 s, extraction time-1 min, centrifugation at 12000× g for 5 min. Conclusions: These results were compared with microwave-assisted extraction procedures. G. purpureum and G. verum extracts showed the highest total phenolic and flavonoid content, respectively. The most potent extract in terms of antioxidant capacity was obtained from G. purpureum, whereas the extract obtained from G. album exhibited the strongest inhibitory effect against tyrosinase.

A multidirectional investigation of stem bark extracts of four African plants: HPLC-MS/MS profiling and biological potentials.

The practice of traditional medicine, especially herbal medicine, is still prevalent across the African continent. Yet, their in-depth pharmacological and chemical exploitation by the scientific community remain a necessity. The aim of the present study was to investigate into the phenolic components, antioxidant, and enzyme-inhibitory activities of three solvent extracts (ethyl acetate, methanol, and water) of the stem bark of four African plant species (Senna siamea, Distemonanthus benthamianus, Harrisonia abyssinica, and Pycnanthus angolensis). It was found that D. benthamianus followed by P. angolensis, displayed the highest DPPH and ABTS scavenging, ferric and cupric reducing, and total antioxidant capacity in the phosphomolybdenum assay. A similar result was observed for AChE, BChE, and tyrosinase inhibition. The two plants also showed comparable α-amylase inhibitory effect. On the other hand, H. abyssinica showed high metal chelating and α-glucosidase inhibition. Among the solvents used, the methanol extract seemed to be the most bioactive. In addition, TPC was highest in D. benthamianus (135.33-192.29 mg GAE/g) while P. angolensis was richest in TFC (7.68-12.48 mg RE/g). A range of bioactive compounds were identified in the extracts, with variations observed among the plants. Senna siamea stem bark showed the presence of nine compounds; where flavonoids (e.g. naringenin, kaempferol, dihydroquercetin) were recorded. Genistein (m/z 271.06), procyanidin B (m/z 577.13) and C (m/z 865.19) isomers were common in stem barks extracts of D. benthamianus and P. Angolensis. To conclude, D. benthamianus and P. angolensis can be considered as potential pharmaceutical agents or functional food components that could reduce the risks of oxidative stress-related disorders.

A salting-out assisted liquid-liquid microextraction procedure for determination of cysteine followed by spectrophotometric detection.

A new analytical method for sensitive determination of cysteine based on its interaction with phenazine methosulfate was developed using salting-out liquid-liquid microextraction followed by spectrophotometric detection. The mechanism of the reaction was studied and confirmed by Fourier transform infrared and mass spectroscopy. Experimental parameters affecting the extraction efficiency were investigated and under the optimal conditions, good linearity was observed in the range 0.2 - 6.0 µg mL-1 with a correlation coefficient of 0.9972. The limit of detection and limit of quantification were found to be 0.07 and 0.21 µg mL -1, respectively. The enrichment factor was 25. The developed methodology was applied for analysis of cysteine in food supplements. The obtained data were in good agreement with LC-MS/MS analysis.

HPLC-MS/MS chemical characterization and biological properties of Origanum onites extracts: a recent insight.

This study investigated into the phytochemical profile and biological properties of extracts (methanol and aqueous) of Origanum onites based on the antioxidant, enzyme inhibitory, and antibacterial activities. The aqueous extract exhibited higher antioxidant activities in the 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS), ferric reducing antioxidant power, cupric reducing antioxidant capacity, phosphomolybdenum, and metal chelating assays, compared to the methanol extract. In contrast, the methanol extract was the most effective inhibitor of acetylcholinesterase, butyrylcholinesterase, tyrosinase, α-amylase, and α-glucosidase. The methanol extract also showed higher antibacterial activity with highest inhibition against Escherichia coli (MIC = 6.25 mg/mL). The total phenolic content was higher in the aqueous extract while the methanol extract possessed higher total flavonoid content. A total of 28 and 18 compounds (belonging to polyphenols, flavonoids, terpenoids, and ester classes) were identified from the methanol and water extracts, respectively. These findings suggest that O. onites could be helpful in the management of oxidative stress-associated diseases including diabetes and neurodegenerative complications. Abbreviations: ABTS: 2,2'-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid; ACAE: acarbose equivalent; AChE: acetylcholinesterase; AD: Alzheimer's disease; BChE: butyrylcholinesterase; CUPRAC: cupric reducing antioxidant capacity; DPPH: 1,1-diphenyl-2-picrylhydrazyl; EDTAE: EDTA equivalent; FRAP: ferric reducing antioxidant power; GAE: gallic acid equivalent; GALAE: galatamine equivalent; HPLC: high performance liquid chromatography; KAE: kojic acid equivalent; RE: rutin equivalents; TE: trolox equivalent; TPC: total phenolic content; TFC: total flavonoid content.

A comprehensive appraisal on Crocus chrysanthus (Herb.) Herb. flower extracts with HPLC-MS/MS profiles, antioxidant and enzyme inhibitory properties.

In the quest for new drugs of herbal origin, the ethyl acetate (EAE), methanol (ME), and water (WE) extracts of Crocus chrysanthus (Herb.) Herb. flowers were analyzed for their polyphenolic composition, antioxidant, and enzyme inhibitory potential. WE showed the highest antioxidant activities in all assays including metal chelating, phosphomolybdenum, FRAP, CUPRAC, ABTS and DPPH. EAE was the most effective enzyme inhibitor, exhibiting the highest inhibition against some enzymes linked to Alzheimer's disease (cholinesterases), diabetes mellitus (α-glucosidase and α-amylase) and hyperpigmentation problems (tyrosinase). The highest total phenolics (34.99 mg GAE/g) and flavonoids content (77.58 mg RE/g) were observed in WE and ME, respectively. Eight compounds were identified in EAE, 24 in ME, and 15 in WE. Kaempferol 3-O glucoside was found in all extracts. In conclusion, C. chrysanthus flowers can be suggested as a source of bioactive components with potential use against chronic disorders caused by oxidative stress. Future in-depth studies are recommended to determine the biological effects of isolated compounds from C. chrysanthus to identify the main compounds modulating the observed activities.

Characterization of phytochemical components of Ferula halophila extracts using HPLC-MS/MS and their pharmacological potentials: a multi-functional insight.

The inhibitory action of F. halophila extracts (acetone, chloroform, and methanol) against key enzymes linked to diabetes (α-amylase, α-glucosidase), cognitive functions (acetyl cholinesterase (AChE), butyryl cholinesterase (BChE)), and hyperpigmentation (tyrosinase) was assessed. The mutagenic/antimutagenic activities were assessed and the phytochemical profile established by HPLC-MS/MS. The acetone extract showed the highest phenolic (55.22 mg GAE/g extract) and flavonoid (34.52 mg RE/g extract) contents. The chloroform extract was a potent inhibitor of cholinesterases (4.86 and 6.13 mg GALAE/g extract, against AChE and BChE, respectively). Cinnamic acid derivatives (methyl cinnamate, ferulic acid, methoxycinnamic acid isomer) were identified in the chloroform extract. Methanol extract showed potent inhibitory action against tyrosinase (137.63 mg KAE/g extract) and glucosidase (43.02 mmol ACAE/g extract). The chloroform extract (32.07 mg EDTAE/g extract) showed potent metal chelating potential. The neuroprotective action of the chloroform extract might be attributed to the metal chelating action coupled by the cholinesterase inhibitory potential. F. halophila showed no mutagenic capacity. When combined with 2-aminoflouren and 2-aminoanthracene, the acetone and chloroform extracts revealed excellent antimutagenicity in the presence of metabolic activation enzymes for Salmonella typhimurium TA98 and TA100 strains. The observed inhibitory effects of F. halophila against the studied enzyme suggest that this plant could be a promising source of bioactive phytochemicals for the management of clinical conditions.

Application of liquid-liquid microextraction for the effective separation and simultaneous determination of 11 pharmaceuticals in wastewater samples using high-performance liquid chromatography with tandem mass spectrometry.

A dispersive liquid-liquid microextraction method for the simultaneous determination of 11 pharmaceuticals has been developed. The method is based on a microextraction procedure applied to wastewater samples from different regions of Hungary followed by high-performance liquid chromatography with mass spectrometry. The effect of the nature of the extractant, dispersive solvent, different additives, and extraction time were examined on the extraction efficiently of the dispersive liquid-liquid microextraction method. Under optimal conditions, the linearity for determining the pharmaceuticals was in the range of 1-500 ng/mL, with the correlation coefficients ranging from 0.9922 to 0.9995. The limits of detection and limits of quantification were in the range of 0.31-6.65 and 0.93-22.18 ng/mL, respectively.

Development of novel techniques to extract phenolic compounds from Romanian cultivars of Prunus domestica L. and their biological properties.

In the present work, fourteen cultivars of Prunus domestica were analyzed to investigate their phenolic pattern with the purpose of using the leaves as potential resources of bioactive compounds in the pharmaceutical and food industry. Microwave-assisted extraction (MAE), dispersive liquid-liquid microextraction and sugaring-out liquid-liquid extraction techniques were optimized in order to obtain an exhaustive multi-component panel of phenolic compounds. The best phenolic-enriched recovery was achieved using MAE in water:methanol (30:70), and this procedure was further applied for quantitative analysis of phenolic compounds in real samples. In order to prove the safeness of these extracts, the biological potential of the Prunus cultivars was tested by several in vitro antioxidant and enzyme inhibitory assays. Moreover, their cytotoxicity was evaluated on human gingival fibroblasts (HGFs), and in most of the cases the treatment with different concentrations of extracts didn't show cytotoxicity up to 500 µg/mL. Only 'Carpatin' and 'Minerva' cultivars, at 250 and 500 µg/mL, reduced partially cell viability of HGFs population. Noteworthy, Centenar cultivar was the most active for the α-glucosidase inhibition (6.77 mmolACAE/g extract), whereas Ialomița cultivar showed the best antityrosinase activity (23.07 mgKAE/g extract). Overall, leaves of P. domestica represent a rich alternative source of bioactive compounds.

Use of Innovative (Micro)Extraction Techniques to Characterise Harpagophytum procumbens Root and its Commercial Food Supplements.

INTRODUCTION: For the determination of harpagoside and the wide phenolic pattern in Harpagophytum procumbens root and its commercial food supplements, dispersive liquid-liquid microextraction (DLLME), ultrasound-assisted DLLME (UA-DLLME), and sugaring-out liquid-liquid extraction (SULLE) were tested and compared. OBJECTIVES: In order to optimise the extraction efficiency, DLLME and UA-DLLME were performed in different solvents (water and aqueous solutions of glucose, ß-cyclodextrin, (2-hydroxypropyl)-ß-cyclodextrin, sodium chloride, natural deep eutectic solvent, and ionic liquid). MATERIAL AND METHODS: The plant material was ground and sieved to obtain a uniform granulometry before extraction. Commercial food supplements, containing H. procumbens are commercially available in Italy. RESULTS: The most effective sodium chloride-aided-DLLME was then optimised and applied for analyses followed by HPLC-PDA. For comparison, microwave-assisted extraction was performed using the same solvents and the best results were obtained using 1% of ß-cyclodextrin or 15% of sodium chloride. CONCLUSION: All commercial samples respected the European Pharmacopoeia monograph for this plant material, showing a harpagoside content ≥ 1.2%. Copyright © 2017 John Wiley & Sons, Ltd.