Automated workflow utilizing saponification and improved epoxidation for the sensitive determination of mineral oil saturated and aromatic hydrocarbons in edible oils and fats.

Refined edible oils and fats are known to contain olefins resisting the typical epoxidation used for the sample preparation of mineral oil saturated and aromatic hydrocarbons (MOSH and MOAH). These olefins can be misinterpreted as MOAH and are therefore an important reason for inconsistent results between laboratories. Collaborative trials confirm this assumption for low MOAH contents near the quantitation limits regularly. In the scope of this work, a new epoxidation approach was developed. Persistent olefins in refined oils could be successfully epoxidized with performic acid. The reaction kinetics was investigated using model substances for biogenic olefins and MOAH. It was rationalized why certain olefins resist epoxidation and which MOAH can potentially get lost. A prominent peak cluster in the MOAH fraction of refined palm oils could be identified by means of GC-MS and explained why it cannot be epoxidized. Based upon this, an automated and streamlined workflow for sample preparation and analysis was composed tackling major problems identified in previously published methods. Optimized and miniaturized saponification, extraction, epoxidation, and enrichment paired with online LC-GC-FID led to a robust method that was tested and validated for edible oils and fats (RSDR < 7% for MOSH and MOAH at values of 14.9 and 2.1 mg/kg, respectively). Due to increased sample amount and minimized blank values, quantitation limits below 1 mg/kg for MOSH and MOAH were achieved. The trueness of the method was verified by analyzing collaborative trial samples.

Molecular signatures of organic particulates as tracers of emission sources.

Chemical signature of airborne particulates and deposition dusts is subject of study since decades. Usually, three complementary composition markers are investigated, namely, (i) specific organic compounds; (ii) concentration ratios between congeners, and (iii) percent distributions of homologs. Due to its intrinsic limits (e.g., variability depending on decomposition and gas/particle equilibrium), the identification of pollution sources based on molecular signatures results overall restricted to qualitative purposes. Nevertheless, chemical fingerprints allow drawing preliminary information, suitable for successfully approaching multivariate analysis and valuing the relative importance of sources. Here, the state-of-the-art is presented about the molecular fingerprints of non-polar aliphatic, polyaromatic (PAHs, nitro-PAHs), and polar (fatty acids, organic halides, polysaccharides) compounds in emissions. Special concern was addressed to alkenes and alkanes with carbon numbers ranging from 12 to 23 and ≥ 24, which displayed distinct relative abundances in petrol-derived spills and exhausts, emissions from microorganisms, high vegetation, and sediments. Long-chain alkanes associated with tobacco smoke were characterized by a peculiar iso/anteiso/normal homolog fingerprint and by n-hentriacontane percentages higher than elsewhere. Several concentration ratios of PAHs were identified as diagnostic of the type of emission, and the sources of uncertainty were elucidated. Despite extensive investigations conducted so far, the origin of uncommon molecular fingerprints, e.g., alkane/alkene relationships in deposition dusts and airborne particles, remains quite unclear. Polar organics resulted scarcely investigated for pollution apportioning purposes, though they looked as indicative of the nature of sources. Finally, the role of humans and living organisms as actual emitters of chemicals seems to need concern in the future.

A robust high-throughput fluorescent polarization assay for the evaluation and screening of SARS-CoV-2 fusion inhibitors.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a serious threat to global health. One attractive antiviral target is the membrane fusion mechanism employed by the virus to gain access to the host cell. Here we report a robust protein-based fluorescent polarization assay, that mimicking the formation of the six-helix bundle (6-HB) process during the membrane fusion, for the evaluation and screening of SARS-CoV-2 fusion Inhibitors. The IC50 of known inhibitors, HR2P, EK1, and Salvianolic acid C (Sal-C) were measured to be 6.1 nM, 2.5 nM, and 8.9 µM respectively. In addition, we found Sal-A has a slightly lower IC50 (3.9 µM) than Sal-C. Interestingly, simple caffeic acid can also disrupt the formation of 6-HB with a sub-mM concentration. Pilot high throughput screening (HTS) of a small marine natural product library validates the assay with a Z' factor close to 0.8. We envision the current assay provides a convenient way to screen SARS-CoV-2 fusion inhibitors and assess their binding affinity.

Effect of Leaf Maturity on Host Habitat Location by the Egg-Larval Parasitoid Ascogaster reticulata.

Adoxophyes honmai, a serious pest of tea plants, prefers to lay eggs on mature tea leaves rather than young leaves. Here, we examined a hypothesis that Ascogaster reticulata, an egg-larval parasitoid of A. honmai, increases the likelihood of encountering host egg masses by searching mature tea leaves when host-derived cues are not available. In a dual-choice bioassay using a four-arm olfactometer, A. reticulata preferred odor from intact, mature leaves versus young leaves. Based on volatile analysis with gas chromatography-mass spectrometry (GC-MS), we identified 5 and 10 compounds from mature and young leaf volatiles, respectively. The 5 components in the extract from intact mature leaves included (Z)-3-hexenyl acetate, (E)-ß-ocimene, linalool, (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), and methyl salicylate. When each individual compound, or quaternary and quintenary blends of them, ratios of which were adjusted to match those of mature leaf volatiles, were provided, parasitoids preferred the full mixture and the quaternary blend devoid of DMNT to the solvent control. Methyl salicylate, one of the components of preferred blends, was not detected among young leaf volatiles. We concluded that the volatile composition of tea leaves changes, depending on their maturity, and that this composition affects foraging behavior of the parasitoid, which is closely related to the host herbivore's oviposition preference.

Chemical composition of essential oils from the underground parts of Glycyrrhiza echinata L. accessions growing wild in Northern Iran.

Glycyrrhiza echinata L. is a perennial plant of considerable commercial importance in medicine, pharmaceuticals, cosmetics, and tobacco industries and the production of food additives as flavoring and sweetening agent. This study, variation on composition and yield of essential oil from the underground parts of five accessions of G. echinata growing wild in northern Iran was investigated. Results showed high variability for the main constituents of essential oil among accessions of G. echinata. The presence of α-pinene and myrcene in two accessions suggests that these volatiles could serve as chemotaxonomic markers and also might be considered as potentially relevant for taste. The presence of ß-caryophyllene and α- caryophyllene in three accessions could be strong potential for being used in medical applications (anticancer and analgesic properties). Generally, the essential oils from G. echinata and other species could be very competitive targets for phytochemical and food studies.

Differential distribution of characteristic constituents in root, stem and leaf tissues of Salvia miltiorrhiza using MALDI mass spectrometry imaging.

Segmentation-quantification is the most commonly used method for studying the tissue distribution of bioactive constituents in plant, but this method would bring uncontrollable pollution, compound migration and denaturation. Mass spectrometry imaging (MSI), as a new method developed in the past 20 years, has high sensitivity, high spatial resolution, high degree of visualization, and low risk of contamination and degeneration when studying tissue distribution of compounds. For the first time we applied matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) to tissue distribution of characteristic constituents of the medicinal plant Salvia miltiorrhiza. From the collected data, we found the regional differences in root, stem, and leaf tissues, and the ion information with differential distribution characteristics. We also identified 18 bioactive constituents in S. miltiorrhiza with their spatial distribution information. In addition, the plant was divided into five parts, and the identified compounds were analyzed for differences between tissues using LC-MS, which results verified those found from the MSI. It is figured out that MALDI-MSI can be reliably applied to the differential distribution of salvianolic acids and tanshinones.

Polyphenol composition and biological activity of Thymus citriodorus and Thymus vulgaris: Comparison with endemic Iberian Thymus species.

The polyphenol compositions of Thymus × citriodorus and Thymus vulgaris extracts as obtained by exhaustive hydroethanolic (HE) extraction and aqueous decoction (AD) were compared. In addition, their compositions and bioactivities were compared to those of Thymus pulegioides and Thymus mastichina, grown under the same edaphoclimatic conditions, and Thymus carnosus. Rosmarinic acid was the most abundant polyphenol followed by luteolin-hexuronide, salvianolic acids I and K. Cluster analysis suggests a similarity of the polyphenol composition of T. citriodorus and T. vulgaris. A significant antioxidant activity was observed and correlated with their polyphenol levels. The same being observed for the higher anti-proliferative activity/cytotoxicity of HE extracts on Caco-2 and HepG2 cells as compared to AD extracts. Significant association between the total phenolic compounds with the anti-proliferative activity, for both cell lines, was observed. These results support the importance of salvianolic acids levels in Thymus extracts and their in vitro anti-proliferative/cytotoxic activities.

Polymethoxy-1-Alkenes Screening of Chlorella and Spirulina Food Supplements Coupled with In Vivo Toxicity Studies.

Selected species of cyanobacteria and green algae have been reported to produce lipophilic polymethoxy-1-alkenes (PMAs) which were shown to exhibit in vivo teratogenicity. Considering that information on PMAs in Arthospira sp. (known commercially as Spirulina) and Chlorella sp. cultivated for food supplement production was essentially lacking, the present study screened Chlorella (n = 10) and Spirulina (n = 13) food supplements registered in the European Union. Mass spectrometry analysis of column fractionated extracts was performed. None of the four variants previously reported in some cyanobacteria and green algae, nor any potentially related structures were detected in the studied samples. Since the isolated lipophilic fractions contained various compounds, they were further screened for in vivo teratogenicity in Danio rerio embryo, and for the potential to induce oxidative stress and genotoxicity in the liver and neurotoxicity in the brain of adult zebrafish. None of the tested food supplements had detectable levels of PMAs or any potentially related structures. No teratogenicity was revealed except for spinal curvature induced by fractions obtained from two Chlorella products. Selected fractions revealed cytotoxicity as indicated by an increased level of reactive oxygen species, catalase activity, lipid peroxidation and increased frequency of DNA strand breaks in hepatic tissue. The majority (60%) of Chlorella fractions induced an increase in cholinesterase activity in zebrafish brain homogenate while exposure to 61.5% of Spirulina fractions was associated with its decrease. The present study confirms that Chlorella and Spirulina food supplements are free of teratogenic PMAs, although the observed in vivo toxicities raise questions regarding the quality of selected products.

Chemical and pharmacological research on the polyphenol acids isolated from Danshen: A review of salvianolic acids.

Danshen, the dried root of Salvia miltiorrhiza Bge, is a common medicinal herb in Traditional Chinese Medicine, which has been used for the treatment of a number of diseases for thousands of years. More than 2000 years ago, the Chinese early pharmacy monograph "Shennong Materia Medica" recorded that Danshen could be used for the treatment of gastrointestinal diseases, cardiovascular diseases, certain gynecological diseases, etc. Since then, Danshen has been widely used clinically in many different prescriptions for many different diseases, especially for the treatment of cardiovascular diseases. Nowadays, many pharmacological studies about the water-soluble components from Danshen have been reported, especially salvianolic acids. It turned out that salvianolic acids showed strong anti-lipid peroxidation and anti-thrombic activities, and among them, SalAA and SalAB were the most potent. This review focused on the achievements in research of salvianolic acids regarding their bioactivities and pharmacological effects. These studies not only shed light on the water-soluble active components of Danshen and their mechanisms at the molecular level, but also provided theoretical information for the development of new medicines from Danshen for the treatment of cardiovascular and cerebrovascular diseases, inflammatory diseases, metabolic diseases, etc.

Chemical composition of essential oils from different parts of Protium heptaphyllum (Aubl.) Marchand and their in vitro antibacterial activity.

In Brazilian folk medicine, Protium heptaphyllum is used to treat inflammatory conditions and to hasten wound repair. This paper aims to investigate the chemical composition and the in vitro antibacterial effects of the essential oils (EOs) obtained from P. heptaphyllum leaves and ripe and unripe fruits against a representative panel of oral pathogens. The GC-FID and GC-MS analysis revealed that the major components determined in P. heptaphyllum essential oils were myrcene (59.0%), ß-elemene (17.2%), limonene (12.9%), spathulenol (12.6%), α-cubebene (11.6%), germacrene D (10.6%), trans-nerolidol (9.8%), and α-cadinol (8.8%). The essential oils of the ripe and unripe fruits showed the strongest antibacterial activity against the anaerobic bacteria Prevotella nigrescens (MIC = 50 µg/mL). The leaf essential oil displayed very promising activity against Streptococcus mutans (MIC = 50 µg/mL) and Streptococcus mitis (MIC = 62.5 µg/mL). The antibacterial activity of EOs against oral pathogens is also described for the first time. [Formula: see text].

Chemical composition of the essential oils and headspace volatiles of Ferulago sylvatica (Besser) Reichenb. from Serbia.

Chemical composition of the headspace volatiles and essential oils isolated from different parts of Ferulago sylvatica was determined by GC and GC/MS analyses. The results showed that headspace volatiles obtained from the aerial parts and roots were similar regarding the number of identified compounds and main components. However, essential oils obtained from different plant organs showed significant differences in chemical composition. Myrcene was the most abundant component of the inflorescences and shoots volatiles, while α-pinene make up over 50% of the root volatiles. Only three components were identified in the root essential oil with 2,3,6-trimethyl benzaldehyde (92.7%) as the main component. In the shoots sample the terpenoid fractions represented 56% of the oil, unevenly distributed between monoterpenoids and sesquiterpenoids with germacrene D (32.5%) recognized as the main constituent. On the other hand, more than 94% of the inflorescences oils were monoterpenoids with myrcene as the most abundant contributor (29.2%).

Effect of essential oil from Ageratum fastigiatum on beta-integrin (CD18) expression on human lymphocytes stimulated with phorbol myristate acetate in vitro.

Agareratum fastigiatum is a Brazilian medicinal plant used as anti-inflammaroty and for wound healing by the folk medicine. In vitro and in vivo studies involving A. fastigiatum essential oil (EOAF) showed indications of anti-inflammatory activity, however, its effect on membrane integrins involved on cell migration is still unclear. Hence, it was evaluated in the present study the effect of EOAF on CD18 frequency on human lymphocytes. By using gas chromatography/mass spectrometry it was identified 9 compounds on EOAF: α-pinene; ß-pinene; ß-myrcene; d-limonene; ß-ocimene; sesquiterpenes; α-copaene; 4,8-ß-epóxi-caryophyllene; germacrene and bicyclogermacrene. On in vitro tests, 6.25 × 10-3 and 12.5 × 10-3 µL/mL EOAF reduced CD18 frequency on phorbol-12-myristate-13-acetate (PMA)-stimulated lymphocytes. Such cells were obtained from peripheral blood of healthy volunteers, and were treated or not with EOAF. They were stained with fluorescent anti-CD18 monoclonal antibodies, after 24 hours incubation. Our data corroborates previous findings, indicating a possible anti-inflammatory activity of EOAF.

[Study on application of salvianolic acids reference extract in quality control of Salvia miltiorrhiza and salvianolic acids for injection].

The purpose of this study was to investigate the feasibility of the salvianolic acids reference extract for quality control for Salvia miltiorrhiza and salvianolic acids for injection. An Agilent ZORBAX SB-C18( 4. 6 mm×250 mm,5 µm) column was used with mobile phase consisting of 0. 1% formic acid-water and 0. 1% formic acid-acetonitrile in gradient elution procedure. The column temperature was 30 ℃; the flow rate was 1 m L·min-1; and the detection wavelength was 288 nm. The content of rosmarinic acid,lithospermic acid and salvianolic acid B in S. miltiorrhiza was determined by using the salvianolic acids reference extract as control substance. The content of caffeic acid,salvianolic acid E,rosmarinic acid,lithospermic acid,salvianolic acid B,and salvianolic acid Y in the salvianolic acids for injection was also determined. The linear relationship between chemicals was good( r>0. 998 9),and the injection precision RSD was 0. 30%-0. 90%. The sexual RSD is between 1. 4% and 3. 0%,and the RSD of the reproducibility of the extract is between 2. 1% and 5. 2%. The recovery rate of the three components in S. miltiorrhiza was 96. 80%-99. 20%,and the recovery rate of the six components in salvianolic acids for injection was 88. 90%-107. 5%. The solution of S. miltiorrhiza and salvianolic acids for injection were stable within 48 h. A total of 8 batches of S. miltiorrhiza and injection were determined by the reference extract,and the difference was smaller than that measured by the monomer control. This study preliminarily verified that the salvianolic acids reference extract can be used as a substitute for the monomer control for the quality control of S. miltiorrhiza and salvianolic acids for injection.

The Pattern of Straight Chain Hydrocarbons Released by Yucca Flowers (Asparagaceae).

The hydrocarbon pattern in the floral scent of Yucca species was found to comprise a group of unbranched, mid-chain alkanes, alkenes, and an alkadiene. In Y. reverchonii, highly dominant (Z)-8-heptadecene is accompanied by (6Z,9Z)-6,9-heptadecadiene and heptadecane as minor components and by traces of other saturated and unsaturated hydrocarbons with similar chain length. Some of these volatiles proved to be perceived by the antennae of Tegeticula cassandra (pollinating seed-eater of Yucca) and Prodoxus decipiens (herbivore on Yucca). The possible biosynthesis of the compounds is discussed.

Dual-mixed/CMC model for screening target components from traditional Chinese medicines simultaneously acting on EGFR & FGFR4 receptors.

Radix Salviae Miltiorrhiae (also known as DanShen (DS) in China), a popular herbal drug in traditional Chinese medicine (TCM) for promoting blood circulation and treating blood stasis, has been reported to possess potential anti-tumor effects. The aim of the study was to develop an effective and practical method for screening and identifying bioactive compounds from Radix Salviae Miltiorrhiae. In this work, the epidermal growth factor receptor (EGFR) and fibroblast growth factor receptors 4 (FGFR4) dual-mixed/cell membrane chromatography (CMC) coupled with high performance liquid chromatography-electrospray ionization-ion trap-time of flight-multistage mass spectrum (HPLC-ESI-IT-TOF-MSn) was established and successfully used to identify the active components from Radix Salviae Miltiorrhiae. Salvianolic acid C (SAC), tanshinone I (Tan-I), tanshinone IIA (Tan-IIA), and cryptotanshinone (C-Tan) were identified as bioactive components with EGFR and FGFR4 activities. MTT and kinase assay were performed to investigate inhibitory effects of these compounds against EGFR and FGFR4 cells growth in vitro. Both cell viability and kinase activity showed that cryptotanshinone acting on EGFR receptor and tanshinone IIA acting on FGFR4 receptor. In conclusion, the EGFR & FGFR4 dual-mixed/CMC can simultaneously screen the bioactive components from TCMs that act on both EGFR and FGFR4 receptors, which significantly improve the efficiency of specific bioactive components identification from a complex system.

Schinus molle essential oil as a potential source of bioactive compounds: antifungal and antibacterial properties.

AIMS: The study was focused on the evaluation of antimicrobial activity in vitro of the essential oil (EO) of leaves from Schinus molle against bacteria and fungi of clinical importance in the search for the discovery of new active compounds. METHODS AND RESULTS: The chemical composition of the S. molle EO was determined by gas chromatography/mass spectrometry and its antimicrobial effect was verified by broth microdilution method. The major compounds found were ß-pinene (25·23%), epi-α-cadinol (21·29%), α-pinene (18·72%), myrcene (11·54%) and sabinene (5·02%). The EO showed significant antifungal activity against Paracoccidioides brasiliensis (39·06 µg ml-1 ), weak action against Cryptococcus neoformans (625 µg ml-1 ) and Trichophyton quinckeanum (625 µg ml-1 ) and was inactive against Candida sp. In the analysis of the antibacterial action, the micro-organisms tested did not show sensitivity. CONCLUSIONS: This study showed a promising result of S. molle volatiles against the fungus P. brasiliensis, which causes paracoccidioidomycosis (PCM), a systemic mycosis of great clinical importance in Latin America. SIGNIFICANCE AND IMPACT OF THE STUDY: The results found here are novel and encourage investigations of the compounds present in this EO, which represents a source of molecules with potential use in the treatment of PCM.

Chemical composition and antimicrobial activity of the essential oils from flowers and leaves of Grindelia integrifolia DC.

Essential oils from flowers and leaves of Grindelia integrifolia DC. were investigated for the first time in terms of chemical composition and antimicrobial activity. The GC-FID/MS analysis allowed for the identification of 58 and 72 volatiles, comprising 92.4 and 90.1% of the oils, respectively. The major components of the flower oil were α-pinene (34.9%) and limonene (13.1%), while myrcene (16.9%), spathulenol (12.3%), ß-eudesmol (11.9%) and limonene (10.1%) dominated among the leaf volatiles. The antimicrobial activity, evaluated against 12 selected bacteria and fungus, was found moderate, with the strongest effect of both oils observed against C. albicans (MIC = MBC: 0.63 and 0.31 mg/mL for flower and leaf oil, respectively).

Correlation between macroscopic characteristics and tissue-specific chemical profiling of the root of Salvia miltiorrhiza.

BACKGROUND: Macroscopic identification has been widely used as a convenient method for herbal authentication and quality assessment. However, sensory evaluation heavily relied on personal experience and lacked enough evidence-based validations. PURPOSE: We aim to reveal the correlation between macroscopic characteristics and tissue-specific chemical composition of the root of Salvia miltiorrhiza (SMR), and then develop a rapid method for quality assessment. METHODS: Thirty-two batches of SMR were collected and evaluated. The outer-surface color and diameter as the representative tissue features of SMR were selected as the macroscopic indexes. SMR were then divided into three parts along transverse section as outer bark, middle part and central part, to explore the spatial distribution of chemicals. Outer-surface color information was converted into RGB values, while the diameter data were expressed by mean distance, respectively. Thirteen major components including eight salvianolic acids and five tanshinones in each part were determined by liquid chromatography tandem mass spectrometry. Finally, several mathematical models were established and optimized to evaluate the correlation between outer-surface color, size and chemical distribution. RESULT: All five tanshinones mainly distributed in the outer bark while salvianolic acids were averagely existed among three parts. Correlational studies revealed that the surface color depth was significantly and positively correlated with tanshinone contents in the outer bark, while the size showed poor correlation in any chemicals. A color-oriented model was thus developed for the prediction of tanshinone contents in SMR, and a 9 × 9 standard color chart was created for easily use. CONCLUSION: This study contributes an alternative method for macroscopic features-based quality evaluation of herbs, and also complements some scientific data for traditional knowledge.

A quality marker study on salvianolic acids for injection.

BACKGROUND: The quality of Chinese medicine (CM) has being an active and challenging research area for CM. Prof. Chang-Xiao Liu et al first proposed the concept of quality marker (Q-Marker) for the quality evaluation and control on CM. This article describe the exploratory studies of Q-Marker in salvianolic acids for injection (SAI) based on this new concept. PURPOSE: This study was designed to screen Q-Marker of SAI and establish its quality control method based on the concept of CM Q-Marker. METHODS: Based on the concept of CM Q-Marker, the SAI was investigated for the identification of chemical components and their sources. The pharmacological effects on cerebral ischemia and reperfusion induced injury in rats were also investigated. Furthermore, the target cell extracts and pharmacokinetic studies were conducted to screen Q-Markers. Finally, the fingerprints and determination based on Q-Markers were established to assess the quality of SAI more effectively. RESULTS: Overall, 20 constituents in SAI were identified. It was found that salvianolic acid B (SA-B), rosmarinic acid (RA), lithospermic acid (LA), salvianolic acid D (SA-D) and salvianolic acid Y (SA-Y) are major chemical components of SAI. Based on chemical components identifications, analysis of their sources, target cell extracts and pharmacokinetic studies, four phenolic acids, namely SA-B, RA, LA and SA-D, were screened and determined as effective Q-Markers of SAI. CONCLUSION: This study demonstrated that the described method is a powerful approach for detecting Q-Markers, which can be used as control index for the quality assessment of CM.

Essential oils Constituents of the leaves of Amomum gagnepainii and Amomum repoense.

The chemical constituents identified in the essential oils hydrodistilled from the leaves of Amomum gagnepainii T.L.Wu, K.Larsen and Turland and Amomum repoense Pierre ex Gagnep (Zingiberaceae) of Vietnam origin are reported. The chemical analyses were performed by means of gas chromatography-flame ionisation detector (GC-FID) and gas chromatography coupled with mass spectrometry (GC-MS). The main compounds of A. gagnepainii were farnesyl acetate (18.5%), zerumbone (16.4%) and ß-caryophyllene (10.5%). On the other hand, Amomum repoense comprised of monoterpenes dominated by ß-pinene (33.5%), (E)-ß-ocimene (9.6%), γ-terpinene (9.1%) and α-pinene (8.4%). This is the first report on the essential oils of A. gagnepainii and A. repoense grown in Vietnam or elsewhere.