Method establishment of components identification and content determination of index components in Gantaishu capsules / 中国药房

OBJECTIVE To establish methods to identify the chemical components of Gantaishu capsule， and determine the contents of 6 index components including glycyrrhizic acid. METHODS The chemical components of Gantaishu capsule were determined by HPLC-TOF/MS； the contents of 6 index components including glycyrrhizic acid were determined by UPLC-MS/MS. RESULTS A total of 41 chemical components were identified in Gantaishu capsules. The linear ranges of glycyrrhizic acid， mangiferin， luteolin， costunolide， oleanolic acid and berberine were 200-10 000 ng/mL（r were all greater than 0.999）. The limits of quantification were 200， 20， 10， 1， 10， 0.5 ng/mL， and the limits of detection were 100， 10， 5， 0.5， 5， 0.25 ng/mL， respectively； RSDs of precision， stability （24 h） and reproducibility tests were all less than 5.0% （n＝6 or n＝3）； the recoveries were 99.05%-101.08% （RSD were all less than 2.0%， n＝6）. The contents of them were 2.42-2.66， 0.85-1.16， 0.35-0.46， 6.18- 6.46， 0.99-1.29， 5.22-5.56 mg/g. CONCLUSIONS The established methods for identification and content determination are rapid and simple， and can be used for the identification of chemical components and the content determination of index components in Gantaishu capsule.

High-Performance Liquid Chromatography-Mass Spectrometry Analysis of Glycoalkaloids from Underexploited Solanum Species and Their Acetylcholinesterase Inhibition Activity.

Solanum glycoalkaloids are gaining increased scientific attention due to their bioactive potential in the defense of plants against pests and pathogens. The comprehensive glycoalkaloid profiling from the leaves, stems, and roots of seven underexploited Solanum species (S. caripense, S. melanocerasum, S. muricatum, S. nigrum, S. quitoense, S. retroflexum, and S. sisymbriifolium) was conducted using high-performance liquid chromatography-time-of-flight mass spectrometry. A total of 51 glycoalkaloids were shared among the studied Solanum species, with concentrations ranging from 7 to 5.63 × 105 ng g-1. Based on the glycoalkaloid composition, plants were separated into two clusters, Cluster 1 (S. melanocerasum, S. nigrum, and S. retroflexum) and Cluster 2 (S. caripense, S. muricatum, S. quitoense, and S. sisymbriifolium). The inhibition activity of glycoalkaloid extracts on acetylcholinesterase showed a half-maximal inhibitory concentration (IC50), ranging from 0.4 (S. nigrum stems) to 344.9 µg mL-1 (S. sisymbriifolium leaves), that was not directly correlated to the total glycoalkaloid contents. This suggests that the composition of glycoalkaloids in the plant extract, rather than the total concentration, is a driver of biological activity. The study provides a framework for the bioprospecting of underexploited Solanum species for exploring bioactive glycoalkaloids and other compounds with potential pesticidal activities for the development of green bioformulation. This is the first comprehensive report on the glycoalkaloid profiles of S. retroflexum.

Chemical characterisation, hypoglycaemic and renoprotective effects of aqueous leaf extract of Limoniastrum guyonianum on fructose-induced metabolic syndrome in rats.

In the present study, we chemically characterised the aqueous leaf extract of Limoniastrum guyonianum by HPLC-TOF/MS and evaluated its effects on fructose-induced metabolic syndrome (MetS) in Wistar rats. MetS groups were given (10% w/v) fructose solution to drink ad libitum for 9 weeks, whereas, normal animals received ordinary water. LG extract was administrated to treated groups by gavage for the last 6 weeks of the experimental period. Fructose feeding as a liquid solution increased body weight, reduced insulin sensitivity, raised blood glucose level and provoked atherogenic dyslipidemia associated with renal oxidative stress and structural damage. Treating MetS rats with LG extract at doses of 100, 200 and 300 mg/kg b.w./day considerably ameliorated the fructose-induced alterations. From this study, it was concluded that aqueous leaf extract of L. guyonianum possesses hypoglycaemic, hypolipidemic, antioxidant and renoprotective abilities against fructose-induced metabolic syndrome in rats.

Rapid Screening and Identification of Antioxidant Active Components in Glycyrrhiza uralensis Decoction Pieces / 中国药房

OBJECTIVE：To establish a method for online detection of antioxidant active components in Glycyrrhiza uroalensis decoction pieces ，and to identify it. METHODS ：The free radical scavenging rate of 1，1-diphenyl-2-trinitrobenzene hydrazine （DPPH）was determined to evaluate the antioxidant activity of G. uralensis decoction pieces. HPLC-UV-DPPH method was used to screen the anti oxidant active components of G. uralensis decoction pieces. HPLC-TOF/MS was used to obtain mass spectrum data and Qualitive Analyst B 06.00 Build 6.0.633.0 software was used to analyze data. Through contrast analysis of UV absorption spectrum，online chromatogram ，mass spectrum information of G. uralensis and the retention time of each compound ，accurate molecular weight ，antioxidant active components were identified by referring to relevant literature. Validation test was also conducted. RESULTS ：DPPH free radical scavenging rate in 8 batches of G. uralensis decoction pieces ranged 55.71％-60.17％. Seven antioxidative active compounds ，including avolomotor ，8-isopentenyl naringin ，yellow lupulin weitone ，isoflavone B ，3′， 4′-dimethoxy3-hydroxy-6-methyl flavone ，glycyrrhizin E and glycyrrhizin H ，could be screened from G. uralensis decoction pieces. After validation ，the peak area of inverted peak generated by online reaction was positively correlated with DPPH free radical scavenging rate. CONCLUSIONS ：Established method is simple and accurate ，and can be used to quickly screen and identify the main antioxidant components of G. uralensis decoction pieces ；the peak area of inverted peak can be used to evaluate the antioxidant active components of G. uralensis decoction pieces.

Insights into the metabolic characteristics of aminopropanediol analogues of SYLs as S1P1 modulators: from structure to metabolism.

SYL927 and SYL930, two aminopropanediol analogues, are novel Sphingosine-1-phosphate receptor 1 (S1P1) modulators with higher selectivity and pharmacological activity compared with FTY720. Although the immunosuppressive activity of SYLs has been well demonstrated, information regarding the metabolic fates of the two chemicals is limited except for the CYP-catalyzed hydroxylation of SYL930. In this study, the biotransformation schemes of the two promising chemicals were investigated and compared using liver microsomes, S9 fractions and recombinant enzymes, and relevant molecular mechanism was primarily demonstrated by ligand-enzyme docking analysis (CDOCKER). As a result, the hydroxylation at alkyl chain on oxazole ring by the action of CYPs was found for both SYLs in vivo. The SULT-catalyzed sulfonation of the hydroxide was observed for SYL927 while the ADH/ALDH-catalyzed oxidation was only discovered for SYL930. The docking analysis suggested that specific non-covalent forces and/or bonding conformations of the hydroxides with biomacromolecules might be involved in the disparate metabolism of SYLs. Exploring the metabolic characteristics will help clarify the substance base for efficacy and safety of the two drugs. The uncovered structure-metabolism relationship in this study may provide an implication for the design and optimization for other S1P modulators.

The Molecular Characterization and Biological Assessment of the Leaves Extracts of Loofah Reveal their Nutraceutical Potential.

BACKGROUND: Luffa cylindrica is a plant that is widely distributed in Africa and Asia and can be grown in regions with tropical or subtropical climates. Few patents dealt with Loofah biological properties, including some functional foods formulated from its leaves. OBJECTIVE: This study aimed to structurally and functionally characterize the bioactive compounds of L. cylindrica leaves grown in two different environments. METHODS: The extracts of L. cylindrica leaves collected from two Tunisian locations: Essouasi (LE), a semi-arid region and Medenine (LM), an arid region, were investigated for their phenolic compounds and fatty acids using HPLC/TOF-MS and GC-MS techniques, respectively. Furthermore, the antioxidant capacity was evaluated with DPPH, Chelating effect, Hydroxyl radical and Superoxide anion scavenging activities while the anticancer activity against HeLa cell lines was assessed using xCELLigence real time cell analyzer and lactate dehydrogenase cytotoxicity assay. RESULTS: The antiproliferative capacity of both extracts was time and dose-dependent, with LE presenting the lowest HeLa cell index (CI = 0.035 ± 0.018, 250 µg/ml). LE also showed the best cytotoxic capacity (56.49 ± 0.8%) and antioxidant potential (IC50 = 54.41 ± 1.12 µg/ml for DPPH and 12.12 ± 0.07 µg/ml for chelating effect). 14 phenolic compounds were detected in LE, with ferulic acid being the major compound (5128.5 ± 4.09 µg Phenols/g), while LM had only 6 phenolics. GCMS analysis showed the presence of omega-3 fatty acids in LE. CONCLUSIONS: Our findings suggest that L. cylindrica leaves, especially when collected from semiarid regions, are promising for formulating nutraceuticals of interest.

First determination of extracellular paralytic shellfish poisoning toxins in the culture medium of toxigenic dinoflagellates by HILIC-HRMS.

Paralytic shellfish poisoning (PSP) toxins have received considerable attention in recent years because of their adverse effects on marine breeding industries and human health. In this study, a reliable method for the analysis of extracellular PSP toxins in the culture medium of marine toxic dinoflagellates was developed for the first time using graphitized carbon black-solid-phase extraction and hydrophilic interaction liquid chromatography-high-resolution mass spectrometry. The limit of quantification of typical PSP toxins in algal culture medium ranged from 0.072 µg/L to 0.151 µg/L under optimal conditions. Satisfactory absolute recoveries (87.5%-102.4%), precision (relative standard deviation ≤ 7.6%), and linearity (R2 ≥ 0.9998) were also achieved. In addition, the proposed method was applied to screen and determine the extracellular PSP toxins of two typical toxigenic dinoflagellates, Alexandrium minutum and Alexandrium tamarense. The total concentrations of the extracellular PSP toxins in A. minutum and A. tamarense over the whole growth period were within 2.0-735.5 and 2.0-19.2 µg/L, respectively. The concentrations of extracellular PSP toxins varied remarkably in the different growth stages of A. minutum and A. tamarense, and the contents of some extracellular PSP toxins were substantially higher than those of intracellular PSP toxins. Therefore, the extracellular PSP toxins released by toxigenic red tide algae cannot be ignored, and their environmental fate, bioavailability, and potential harm to aquatic environment need to be investigated in future studies.

Characterization of ethyl acetate and n-butanol extracts of Cymbopogon schoenanthus and Helianthemum lippii and their effect on the smooth muscle of the rat distal colon.

ETHNOPHARMACOLOGY RELEVANCE: Cymbopogon schoenanthus (C. schoenanthus) and Helianthemum lippii (H. lippii) are Saharan species found in the South West of Algeria, in the region of Bechar. Both plants are used in traditional medicine to treat gastrointestinal disorders. OBJECTIVE: The aim of our study was to characterize the composition of the ethyl acetate (EtOAc) and n-Butanol (n-BuOH) extracts of C. schoenanthus and H. lippii, and to elucidate and compare their effect on the reactivity of the rat distal colon. MAIN METHODS: The plants were macerated in a hydroalcoholic solution. After concentration, the aqueous solutions of the residues were submitted to liquid-liquid extractions to obtain EtOAc and n-BuOH extracts. The phenolic and flavonoid content of the extracts was determined by high performance liquid chromatography coupled with mass spectrometry with a time of flight analyzer (HPLC-TOF/MS). The effect of the extracts was tested on the rat distal colon, namely on the basal tone and on KCl- and Ach-induced precontracted preparations. RESULTS: HPLC-TOF/MS identified 32 phenols and flavonoids in the extracts. The four extracts relaxed the rat distal colon, the effect being noticed on the basal tone and on the KCl- and Ach-induced precontractions. The EtOAc and the n-BuOH extracts of H. lippii decreased the basal tone of the rat distal colon more markedly than the correspondent extracts of C. schoenanthus. Moreover, the n-BuOH extract of C. schoenanthus decreased the basal tone more markedly than the EtOAc extract of this plant but there was no difference between extracts of H. lippii. The EtOAc extracts of both C. schoenanthus and H. lippii totally reverted both the KCl- and the Ach-induced precontraction of the rat distal colon. However, the n-BuOH extracts of the two plants reverted the Ach-precontracted colon but not the colon that has been precontracted with KCl. CONCLUSION: Extracts of H. lippii contain a higher level of phenols compared to the extracts of C. schoenanthus. All extracts of C. schoenanthus and H. lippii caused marked relaxation of the isolated rat distal colon, either when applied directly or when tested over KCl- and Ach-induced precontraction. These results give support to the use of C. shoenanthus and H. lippii in traditional medicine, namely for gastrointestinal diseases.

Identification of key metabolites during cisplatin-induced acute kidney injury using an HPLC-TOF/MS-based non-targeted urine and kidney metabolomics approach in rats.

Kidney injury is a major adverse effect of cisplatin use. Metabolomics has been used to characterize physiological or pathological conditions through identification of metabolites and characterization of the metabolic pathway. Metabolomics profiling could allow for identification of nephrotoxic mechanisms of cisplatin and identification of biomarkers of cisplatin-induced injury. In this study, we performed metabolomics analysis to characterize key changes in metabolite levels during cisplatin-induced acute kidney injury (AKI) in rats, and screened for sensitive biomarkers for early diagnosis using HPLC-TOF/MS. Rats were intraperitoneally injected with 7.5 mg/kg or 15 mg/kg of cisplatin, or normal saline, and 12 h urine and kidney samples were collected after 72 h. Serum biochemical parameters and kidney histological evaluations showed dose-dependent AKI in response to cisplatin. Metabolomics analysis showed that 37 and 35 endogenous metabolite levels changed in rat urine and kidneys, respectively. Seven key metabolic pathways were disrupted, including the tricarboxylic acid cycle (TCA cycle), phenylalanine, tyrosine, and tryptophan biosynthesis, phenylalanine metabolism, glycerophospholipid metabolism, taurine and hypotaurine metabolism, d-glutamine and d-glutamate metabolism, and nicotinate and nicotinamide metabolism. These pathways are involved in energy generation, and amino acid and lipid metabolism, and disruption of these pathways could contribute to oxidative stress injury, inflammation, and cell membrane damage. Furthermore, 11 sensitive metabolites in urine were screened as potential biomarkers of AKI. To validate these biomarkers, we quantified 4 off these biomarkers, and confirmed that levels of these metabolites were altered in urine of rats treated with CDDP.

Qualitative and quantitative assessment of related substances in the Compound Ketoconazole and Clobetasol Propionate Cream by HPLC-TOF-MS and HPLC.

Related substances in pharmaceutical formulations are associated with their safety, efficacy and stability. However, there is no overall study already published on the assessment of related substances in the Compound Ketoconazole and Clobetasol Propionate Cream. In this work, a reliable HPLC-TOF-MS qualitative method was developed for the analysis of related substances in this preparation with a quick and easy extraction procedure. Besides the active pharmaceutical ingredients, two compounds named ketoconazole impurity B' optical isomer and ketoconazole impurity E were identified. Furthermore, a new HPLC method for qualitative and quantitative assessment on related substances and degradation products, which were found in the stability test, was established and validated. The single standard to determine multi-components method was applied in the quantitative analysis, which was an effective way for reducing cost and improving accuracy. This study can provide a creative idea for routine analysis of quality control of the Compound Ketoconazole and Clobetasol Propionate Cream.

A new nutraceutical resource from a rare native plant growing in Turkey and for its spectro-chemical and biological insights: Endemic Diplotaenia bingolensis (Apiaceae).

The present study designed to investigate the quantitative distributions in the secondary metabolites and biological activity of sub-fractions obtained successively from water and methanol: dichloromethane (rate 1:1; v:v) solvent decoction of Diplotenia bingolensis aerial parts. The crude extracts were obtained from the aerial parts of the endemic D. bingolensis species refluxing with water and organic solvents. Sub-fractions of water extract were obtained by successive fractionation of the water extract with hexane (WH), dichloromethane (WD), ethyl acetate (WE) and n-butanol (WB), respectively. Sub-fractions of organic solvent were obtained by fractionation of the organic crude extract with hexane (OH), dichloromethane (OD), ethyl acetate (OE), n-butanol (OB) and water (OW), respectively. The total amount of phenols and flavonoids contained in each sub-fraction was analyzed by UV-VIS spectrophotometer, analysis of lipophilic components by GC-MS spectrometer, and quantitative analysis of hydrophilic components by HPLC-TOF/MS spectrophotometer. Furthermore, the biological activity of each sub-fraction was compared with different antioxidant activities such as DPPH radical scavenging activity and ferric ion reduction capacity. Sub-fraction WD (137.1 ±â€¯2.1 µg QE/mg DI) and OE (127.1 ±â€¯5.2 µg QE/mg DI) in terms of flavonoid content, sub-fraction WD (665.8 ±â€¯47.6 µg GAE/mg DI) and OE (724.6 ±â€¯43.6 µg GAE/mg DI) were the richest isolates in terms of total phenol content. Sub-fractions OH and OD contained linoleic acid (17.0 and 11.0%, respectively) and linolenic acid (22.1 and 18.5%, respectively). It was revealed that sub-fractions were rich in terms of rutin (1.2-47.2 µg HC/mg DI) and chlorogenic acid (0.1-12.1 µg HC/mg DI). Sub-fractions WD and OE were showed the highest DPPH radical scavenging activity with 46.4 ±â€¯1.4 and 47.6 ±â€¯10.0 µg/mL EC50 values, respectively. This study is the first to demonstrate biological insight of the potential antioxidant activity of D. bingolensis. These findings warrant the popular use of the endemic D. bingolensis and highlight the potential of its active constituents in the development of new antioxidative drugs.

Mycotoxins in herbal teas marketed in Latvia and dietary exposure assessment.

The occurrence of 12 mycotoxins has been analysed by liquid chromatography - time of flight mass spectrometry in the batch of 60 herbal teas purchased from drugstores in Latvia. Among the dry tea samples, 90% were positive for one to eight mycotoxins. Enniatin B and deoxynivalenol (DON) were the most frequently detected mycotoxins in 55% and 45% of the samples, respectively. DON reached the highest level, from 129 µg kg-1 in herbal blend to 5,463 µg kg-1 in wormwood tea. Ochratoxin A (OTA) and aflatoxin B1 (AFB1) were found in 10% and 20% of the samples at the concentrations ranged between 2.99-30.3 µg kg-1 and 3.40-23.7 µg kg-1. Studies of the tea infusion process indicated that 32-100% of DON and zearalenone present in dry teas were extracted into the infusions. Dietary exposure assessment was performed, using the determined mycotoxin levels and the available consumption data.

Fructus Gardeniae-induced gastrointestinal injury was associated with the inflammatory response mediated by the disturbance of vitamin B6, phenylalanine, arachidonic acid, taurine and hypotaurine metabolism.

ETHNOPHARMACOLOGICAL RELEVANCE: Fructus Gardenia (FG) is a widely used bitter and cold herb for clearing heat and detoxicating. Currently, toxicity of FG and its relative formula has been reported in many clinical and animal studies. However, no systematic research has been carried out on FG-related gastrointestinal (GI) injury which has been emphasized in China since the Ming Dynasty. AIM OF THE STUDY: The purpose of this article is to investigate whether FG could damage GI and explore the mechanisms involved. MATERIAL AND METHODS: FG was given to male mice by 7-day intragastric administration at average doses of 0.90 g (L group), 1.50 g (M group), and 3.00 g (H group) crude drug/kg FG. Comprehensive understanding of changes in weight, diarrhea degree, stool routine, histomorphology and inflammatory factors of stomach, small intestine, and colon for evaluating the effect of different doses of FG on GI injury. Moreover, metabolomics-based mechanisms exploration of FG on GI injury was carried out via HPLC-Q-TOF/MS analysis on mice urine. RESULTS: High dose FG caused GI injury with serious diarrhea, decreased weight, abnormal stool routine, sever alteration in histomorphology of small intestine and colon (mild change in stomach), and significant change in inflammatory factors. The results of metabolomics suggested that 55 endogenous metabolites dispersed in 21 significantly altered metabolic pathways in 3.00 g/kg crude FG treated mice. The hub metabolites of GI injury were mainly related with vitamin B6 metabolism, phenylalanine metabolism, arachidonic acid metabolism, and taurine and hypotaurine metabolism via correlated network analysis. CONCLUSION: FG affected the normal functions of GI via the regulating a variety of metabolic pathways to an abnormal state, and our results provided a research paradigm for the GI-injury of the relative bitter and cold traditional Chinese medicines.

Qualitative and quantitative assessment of related substances in the Compound Ketoconazole and Clobetasol Propionate Cream by HPLC-TOF-MS and HPLC / 药物分析学报

Related substances in pharmaceutical formulations are associated with their safety, efficacy and stability. However, there is no overall study already published on the assessment of related substances in the Compound Ketoconazole and Clobetasol Propionate Cream. In this work, a reliable HPLC-TOF-MS qua-litative method was developed for the analysis of related substances in this preparation with a quick and easy extraction procedure. Besides the active pharmaceutical ingredients, two compounds named ke-toconazole impurity B′ optical isomer and ketoconazole impurity E were identified. Furthermore, a new HPLC method for qualitative and quantitative assessment on related substances and degradation pro-ducts, which were found in the stability test, was established and validated. The single standard to determine multi-components method was applied in the quantitative analysis, which was an effective way for reducing cost and improving accuracy. This study can provide a creative idea for routine analysis of quality control of the Compound Ketoconazole and Clobetasol Propionate Cream.

Phenolic Profile, Essential Oil Composition and Bioactivity of Lasia spinosa (L) Thwaites

Abstract Lasia spinosa (L.) Thwaites is a widely used ethnomedicinal plant in Bangladesh. In this study, we investigated phenolic contents, volatile compounds and fatty acids, and essential oil components of extracts prepared from aerial parts of the plant. The main volatile compounds were methyl ester of oleic acid, palmitic acid and stearic acid as determined by GC/MS. Phenolic contents of the extracts were determined qualitatively and quantitatively by HPLC/TOF-MS. Six phenolic compounds (syringic acid, morin, gentistic acid, 4-hydroxybenzoic acid, cinnamic acid, and apigenin) were found in the extracts. GC/MS analysis of steam distilled essential oil showed camphor, α-pinene and δ-3-carene as the main constituents. In DPPH radical scavenging assay, the highest free radical scavenging activity was observed for the methanol extract with an IC50 value of 0.48 ± 0.04 mg/mL, whereas, in metal chelating activity on ferrous ions (Fe2+) assay, the highest chelating activity was observed for hexane extract (IC50 = 0.55 ± 0.08 mg/mL). The extracts and essential oil were tested against five severe human pathogenic bacteria using disc diffusion assay and subsequent MIC values were also determined. All the extracts (except methanol extract) and the essential oil were found to possess potential antimicrobial activity with corresponding inhibition zone and minimum inhibitory concentration (MIC) ranging from 9-23 mm and 62.5-500 µg/mL. This study has been explored the plant Lasia spinosa can be seen as a potential source of biologically active compounds.

Characterization of bioactive compounds of Annona cherimola L. leaves using a combined approach based on HPLC-ESI-TOF-MS and NMR.

Annona cherimola Mill. (cherimoya) has widely been used as food crop. The leaves of this tree possess several health benefits, which are, in general, attributed mainly to its bioactive composition. However, literature concerning a comprehensive characterization based on a combined approach, which consists of nuclear magnetic resonance (NMR) and high-performance liquid chromatography coupled with time-of-flight mass spectrometry (HPLC-TOF-MS), from these leaves is scarce. Thus, the aim of this work was to study the polar profile of full extracts of cherimoya leaves by using these tools. Thus, a total of 77 compounds have been characterized, 12 of which were identified by both techniques. Briefly, 23 compounds were classified as amino acids, organic acids, carbohydrates, cholines, phenolic acid derivatives, and flavonoids by NMR, while 66 metabolites were divided into sugars, amino acids, phenolic acids and derivatives, flavonoids, phenylpropanoids, and other polar compounds by HPLC-TOF-MS. It is worth mentioning that different solvent mixtures were tested and the total phenolic content in the extracts quantified (TPC via HPLC-TOF-MS). The tendency observed was EtOH/water 80/20 (v/v) (17.0 ± 0.2 mg TPC/g leaf dry weight (d.w.)) ≥ acetone/water 70/30 (v/v) (16.1 ± 0.7 mg TPC/g leaf d.w.) > EtOH/water 70/30 (v/v) (14.0 ± 0.3 mg TPC/g leaf d.w.) > acetone/water 80/20 (v/v) (13.5 ± 0.4 mg TPC/g leaf d.w.). Importantly, flavonoids derivatives were between 63 and 76% of the TPC in those extracts. Major compounds were sucrose, glucose (α and ß), and proline, and chlorogenic acid and rutin for NMR and HPLC-TOF-MS, respectively. Graphical abstract The combined use of LC-HRMS and NMR is a potential synergic combination for a comprehensive metabolite composition of cherimoya leaves.

The effects of boiling and fermentation on betalain profiles and antioxidant capacities of red beetroot products.

The aim of study was investigation the impact of boiling and spontaneous fermentation on profile and content of betalains and antioxidant capacity of red beetroot. Betalains were analyzed by micro-HPLC-TOF-MS/MS method, while antioxidant capacity by three in vitro assays. Red beet products were abounded in betalains, with betanin, isobetanin, betanidin and vulgaxanthin I predominating among twenty two pigments identified. Boiling and fermentation of red beet reduced the content of betalains by 51-61% and 61-88%, respectively, however, this decline was limited when the peel was present. Microbial activity and softening of the matrix induced by the fermentation process led to the release of betalains responsible for strong antioxidant capacity of the juice formed. Generally, the treatment applied maintained and/or lowered antioxidant capacity of red beet materials obtained. Our findings indicate that red beet-derived products are valuable source of betalains, with their profile, content and antioxidant capacity being modulated by processes applied.

Phytochemical Screening, Antiproliferative and Antioxidant Properties of Various Extracts from Endemic Origanum acutidens.

AIM AND OBJECTIVE: Origanum acutidens (Hand.-Mazz.) Ietsw. is an endemic and perennial plant grown mainly in East Anatolia. Recently, natural plant products have attracted interest due to their safety and therapeutic effects. Therefore, the aim of this study was to investigate phytochemical contents and biological effects of Origanum acutidens. MATERIALS AND METHODS: The aerial parts of O. acutidens were extracted with water, ethyl acetate, nbutanol, and methanol/chloroform solvents. Phenolic compounds and other constituents of the extracts were analyzed by HPLC/TOF-MS. The Ethyl Acetate extract (EA) was fractionated by flash chromatography. The extracts and fractions were investigated for their antiproliferative activities on human cervical adenocarcinoma (HeLa) cell line by using BrdU ELISA assay. Antioxidant activities of the extracts and fractions were evaluated by complementary test systems, namely determination of total phenolic contents, metal chelating ability and DPPH radical scavenging assay. RESULTS: Among the extracts, Ethyl Acetate extract (EA) exhibited the highest antiproliferative activity (IC50 = 15.71 ± 0.04 µg/mL) on HeLa cells. It was therefore fractionated by flash chromatography to obtain 10 fractions which were investigated for their phenolic compounds and bioactivities. Rosmarinic acid was determined as the major component of EA and its fractions. EA exhibited higher antiproliferative activity against HeLa cell line than its fractions and 5-fluorouracil (5-FU) at the concentration of 100 µg/mL. EA and its fractions (F10, F6, F4, F7, F3, and F2) displayed higher radical scavenging activity compared to Butylated Hydroxytoluene (BHT). These effects may be attributed to the presence of rosmarinic acid in EA and its active fractions. CONCLUSION: This study demonstrated that O. acutidens is an essential natural source of polyphenols and a potent natural antioxidant and antiproliferative agent for food and pharmaceutical industries.

Analysis of constituents from different parts of Rhodomyrtus tomentosa by HPLC-TOF-MS / 中草药

Objective To compare chemical composition in the different parts (leaf, branch, and fruit) of Rhodomyrtus tomentosa, and to study the chemical constituents from fruits of R. tomentosa. Methods High performance liquid chromatography/time-of-flight mass spectrometry (HPLC-TOF-MS) method was executed to analyze the samples. Principle component analysis (PCA) and partial minimum variance discriminant analysis (OPLS-DA) in MassLynx XS software were used to analyze the obtained data. The chemical constituents of fruits were isolated and purified by column chromatography, including silica gel, Sephedex LH-20 and re-HPLC, and the structures were elucidated based on their NMR and MS data. Results The PCA results indicated that the constituents existed in leaf were significantly different from those in branch and fruit, while constituents in branch and fruit were similar. Furthermore, based on OPLS-DA, combined with chromatographic retention regulation, accurate molecular mass, isotopic matching and literature searching, four marker compounds from leaves had been found and identified as myricitrin (1), myricitrin-3-O-L-furanoarabinoside (2), iridin (3), and 3,3’-didemethyl-9-oxo-pinoresinol (4). Besides, five compounds were isolated from fruits and identified as maslinic acid (5), ethyl gallate (6), gallic acid (7), resveratrol (8), and piceatannol (9). Conclusion This research provides an effective strategy for analyzing chemical difference from different parts of R. tomentosa, which can be applied to study the chemical difference from different parts of other species.

Development of a comprehensive screening method for more than 300 organic chemicals in water samples using a combination of solid-phase extraction and liquid chromatography-time-of-flight-mass spectrometry.

A comprehensive screening method for 311 organic compounds with a wide range of physicochemical properties (log Pow -2.2-8.53) in water samples was developed by combining solid-phase extraction with liquid chromatography-high-resolution time-of-flight mass spectrometry. Method optimization using 128 pesticides revealed that tandem extraction with styrene-divinylbenzene polymer and activated carbon solid-phase extraction cartridges at pH 7.0 was optimal. The developed screening method was able to extract 190 model compounds with average recovery of 80.8% and average relative standard deviations (RSD) of 13.5% from spiked reagent water at 0.20 µg L-1, and 87.1% recovery and 10.8% RSD at 0.05 µg L-1. Spike-recovery testing (0.20 µg L-1) using real sewage treatment plant effluents resulted in an average recovery and average RSD of 190 model compounds of 77.4 and 13.1%, respectively. The method was applied to the influent and effluent of five sewage treatment plants in Kitakyushu, Japan, with 29 out of 311 analytes being observed at least once. The results showed that this method can screen for a large number of chemicals with a wide range of physicochemical properties quickly and at low operational cost, something that is difficult to achieve using conventional analytical methods. This method will find utility in target screening of hazardous chemicals with a high risk in environmental waters, and for confirming the safety of water after environmental incidents.