The remains of black henbane (Hyoscyamus niger) are relatively common at archaeological sites as it grows naturally around settlements in north-western Europe. All parts of the plant may be used as a medicine or a narcotic but its natural prevalence in built environments makes it difficult to interpret any intentionality behind its presence in the archaeological record. Evidence of the deliberate collection and use of black henbane seeds in the Roman Netherlands is presented here for the first time. Examination of Classical texts and interrogation of the archaeobotanical data allow the authors to place the discovery at Houten-Castellum of a hollowed bone containing hundreds of black henbane seeds within the context of the wider Roman understanding of the plant and its properties.

In Mars-analogous hyperarid soils of the Atacama Desert, phospholipid fatty acids (PLFAs) as the main component of biomembrane play a role in reliably determining viable microbes. PLFA analyses illustrated a rise of the microbial abundance (from 5.0 × 106 to 4.2 × 107 cells g−1) and biodiversity (from 7 to 15 different individual PLFAs) from the north hyperarid core of the Atacama Desert to the southern arid region. Abundant cyclopropyl PLFAs (47.2 ± 4.6%) suggested the resistance to oligotrophic and hypersaline environments by Atacama microbial communities. The southernmost arid site had the highest proportion (8.7%) of eukaryotic and fungal lipid biomarkers. Different precipitations (ranging from 0.7 to 2 mm year−1) in the hyperarid core exerted different effects on microbial biomass, PLFA diversity, bacteria and microeukaryotes. By principal component analysis (cumulative 74.6% of variance), the dominance of PLFA hydroxylation was associated with the microbial viability; bacteria rich in C16:0, C18:0 and C16:1ω9 favoured higher soil conductivity and nitrate; and other PLFAs contributed more to the organic content. Additionally by comparing the ratios of PLFAs to well-preserved organics (e.g., mineral-bound organic carbon and microbial phosphorus), I found that the degradation of PLFAs decreased to a minimum when the mean annual precipitation is lower than 2 mm. These findings may further specify identifiable biomarkers on Mars, if potentially extant Martian microbes possess comparable phospholipid membrane structure.

Previous scientific studies have listed chemical compounds from sea cucumber species. The ability to produce bioactive metabolites derives from their response to various ecological stresses and avoidance of predation. In view of this, the present work aimed to evaluate the antibacterial activity and quantitatively assess the active components of different organs of the sea cucumber Holothuria foskali collected from the north-east of Tunisia. The antibacterial activity of ethyl acetate extracts was tested against Bacillus subtilis, Bacillus cereus, Escherichia coli and Pseudomonas aeruginosa. It was revealed that Escherichia coli and Bacillus subtilis were inhibited by the sea cucumber extracts. To identify the composition of these extracts, Gas Chromatography-Mass Spectroscopy (GC-MS) and Liquid Chromatography-Mass Spectroscopy (LC-MS) methods were used. The GC-MS revealed the presence of 25 major components, the most concentrated ones were characterized as C26H27N5O6 Morphinan-4,5-epoxy-3,6-diol, 6 amino;C17H31F3O2 3-trifluoroacetoxypentadecane; C31H50O2 Stigmasta-5,22-dien-3-ol,acetate; C11H14O2 1-Phthalanol,1,3,3-trimethyl 1-phenyl; C30H48O2 Ergosta-5,22-dien-3-ol,acetate and C25H40O2 (9E)-8-Phenyl-9-octadecenoic acid methyl ester; while the LC-MS revealed the presence of eight molecules, identified as phenolic acid, phytosterol and fat-soluble vitamins. Some molecules are interesting and deserve to be explored for their potential uses as ingredients in functional foods or as raw materials for new drug forms.

To investigate the effects of heat stress on broiler metabolism, we assigned 144 broilers to normal control (NC), heat stress (HS) or pair-fed (PF) groups and then monitored the effects using growth performance, carcass characteristics, biochemical assays and GC-MS-based metabolomics. The up-regulation of cloacal temperature confirmed that our experiment was successful in inducing chronic heat stress. The average daily gain and average daily feed intake of the HS group were significantly lower than those of the NC group, by 28·76 and 18·42 %, respectively (P<0·001), whereas the feed:gain ratio was significantly higher, by 14·49 % (P=0·003), and heat stress also increased leg proportion (P=0·027) and intramuscular fat proportion (P<0·001) and decreased breast proportion (P=0·009). When comparing the HS and NC groups and HS and PF groups, our metabolomics approach identified seventy-eight and thirty-four metabolites, respectively, with significantly different levels (variable importance in the projection values >1 and P<0·05). The greater feed:gain ratio of the HS group was significantly positively correlated with the leg, abdominal fat, subcutaneous fat and intramuscular fat proportions and levels of some free amino acids (proline, l-cysteine, methionine and threonine) but was negatively correlated with breast proportion and levels of some NEFA (stearic acid, arachidonic acid, palmitic acid and oleic acid). These findings indicated that the heat-stressed broilers were in negative energy balance and unable to effectively mobilise fat, thereby resulting in protein decomposition, which subsequently affected growth performance and carcass characteristics.

The relic “the sack of Saint Francesco” has for the first time been investigated by scientific means. The sack is kept at the Franciscan Friary of Folloni near Montella in southern Italy. According to legend, the sack appeared on the doorstep of the Friary in the winter of 1224 containing bread sent from St Francesco (St Francis of Assisi), who at that time was in France. The bread was allegedly brought to the friary by an angel. We analyzed samples of the sack to obtain a radiocarbon (14C) date and to search for any remaining traces of bread. The 14C date yielded a calibrated age range of AD 1220–1295 (2σ), which places the textile in the right timeframe according to the legend. Chemical analysis by gas-chromatography with mass spectrometric detection (GC-MS) revealed the presence of ergosterol (5, 7, 22-ergostatrien-3b-ol), a known biomarker of brewing, baking, or agriculture. In this paper we have further substantiated the validity of ergosterol as a biomarker for the past presence of bread. It appears that there is a fine correspondence between the Franciscan legend and the two most decisive scientific methods relevant for analyzing the sack. Although it is not proof, our analysis shows that the sack indeed could be authentic.

Sea food is very rich in lipids. The brown shrimp, Crangon crangon is a most popular and very valuable taxon in the White Sea, in the Mediterranean and in the Black Seas. The aim of this study was to determine the seasonal variations of lipids in C. crangon muscle tissue using gas chromatography–mass spectrometry. The lipids were separated into groups: neutral lipids (triacylglycerols, free fatty acids, sterols) and polar lipids (phospholipids), by high performance liquid chromatography with a laser light-scattering detector. Fatty acids were identified using the GC-MS technique. The mainly fatty acids were 16:0, 18:0 (saturated FAs), 16:1, 18:1 (monounsaturated FAs), 20:4 (ARA), 20:5 (EPA) and 22:6 (DHA). The largest amounts of fatty acids in the muscle were observed in spring; these were the result of collecting food after winter and before reproduction. The muscle lipid content was 32.2 ± 1.8 mg g−1. The summer was the poorest season and the lipid value was 7.7 ±0.4 mg g−1. The levels of muscle neutral lipids (NL) oscillated between 80% (autumn) and 90% (spring). The temperature and salinity has a significant influence on content and profile of fatty acids. This work will help to understand the biology, the seasonal variation in lipid mass, lipid classes content and fatty acids profile in the abdominal muscle of C. crangon.

As gut immaturity precludes full enteral feeding, very low birth weight (VLBW) preterm infants receive parenteral nutrition (PN) during the first few weeks of life. Weaning VLBW infants off PN, however, is a top priority since PN is associated with a high risk of complications. The decision making is purely empirical, as there is currently no suitable index of gastrointestinal (GI) maturity. Plasma citrulline concentration is considered an index of GI function in conditions such as short-bowel syndrome and coeliac disease in adults. To identify the factors determining urinary citrulline excretion, and determine whether urinary citrulline excretion could be used as a non-invasive index of GI tolerance to enteral feeding, nutritional intake and urinary citrulline were monitored bi-weekly in forty-seven preterm infants < 1500 g (interquartiles 880–1320 g), during their stay in the Neonatology unit. Median urinary citrulline was 24·7 μmol/mmol creatinine (14·5–38·6 μmol/mmol creatinine). No relationship was observed with the percentage of energy tolerated enterally. In multivariate regression analysis, weak correlations were found with post-conceptional age (P = 0·001), parenteral amino acid supply (P = 0·001) and the daily volume of enteral mixture administered (P = 0·043). A significant correlation was found with urinary nitrite+nitrate excretion (r 0·47; P < 0·001). We conclude that in preterm infants: (1) one of the major determinants of urinary citrulline may be the biosynthesis of citrulline from arginine by NO-synthase; (2) urinary citrulline cannot be used to predict GI tolerance. This is consistent with the observations that, in neonatal gut, citrulline is converted to arginine in situ rather than exported towards the kidneys as observed in adults.

The ecologically important lichen Cladonia stellaris forms thick carpets in boreal forest floors. In addition to affecting temperature and water conditions in the soil underneath, the secondary metabolites formed by the lichen layer are of ecological interest. In this paper, we investigated the distribution of lichen acids in C. stellaris collected at different latitudes in Finland and developed methods to quantify the two optical enantiomers of usnic acid separately. The lichen extracts were analysed by high-performance liquid chromatography (HPLC) with UV and mass spectrometric (MS) detection and by gas chromatography with flame ionization (GC-FID) and MS detection. Usnic acid and perlatolic acid were quantified using GC-FID. The concentration of usnic acid in the top 20 mm of the lichen thallus ranged from 0·48–3·08% of dry weight, and that of perlatolic acid from 0·08–0·54%. The enantiomeric composition of usnic acid was determined using a chiral HPLC column coupled to an electrospray ionization-tandem mass spectrometer. (−)-Usnic acid was found to be the predominating enantiomer in all extracts; the proportion of (+)-usnic acid ranged from 0·4%–10·0%. Olivetoric acid methyl ester, diphenylmethanol, and 5-pentylresorcinol were identified, and several other olivetoric acid-type compounds were tentatively identified in the extracts.

The strengths and limitations of existing mass spectrometry methods for metabolite detection and identification are discussed. A brief review is made of the methods available for quenching and extraction of cells or organisms prior to instrumental analysis. The techniques available for carrying out mass spectrometry-based profiling of metabolomes are discussed using the analysis of extracts from trypanosomes to illustrate various points regarding methods of separation and mass spectrometric analysis. The advantages of hydrophilic interaction chromatography (HILIC) for the analysis of polar metabolites are discussed. The challenges of data processing are outlined and illustrated using the example of ThermoFisher's Sieve software. The existing literature on applications of mass spectrometry to the profiling of parasite metabolomes is reviewed.

The composition of essential oils obtained from leaves of Acorus calamus growing wild in 19 different locations was analysed by gas chromatography and gas chromatography–mass spectrometry. In total, 84 constituents have been identified, representing at least 86% of the total essential oil. The essential oils were dominated by the presence of phenolic compounds: (Z)-asarone (15.7–25.5%) and (Z)-methyl isoeugenol (2.0–4.9%). Other identified major components were (E)-caryophyllene, α-humulene, germacrene, linalool, camphor and isoborneol. All oils were found to have a similar composition. However, a hierarchical cluster analysis of the quantitative data revealed significant differences between some accessions. Minimal inhibitory concentrations (MIC) of four selected leaf essential oils were determined by a screening test with the broth dilution method. Of the 17 species of fungi, bacteria and yeasts tested, Mycobacterium ssp. and Bacillus subtilis were most susceptible to essential oils. Oils displayed low anti-fungal activity and their inhibitory activity was lower than the standard antibiotics. Oils differed in their activities against the various microorganisms tested.

Ewe milk fat from five different herds was studied to determine the content of conjugated linoleic acid (CLA) isomers. Research was carried out by combining gas chromatography-mass spectrometry (GC-MS) of fatty acid methyl esters (FAME) and 4,4-dimethyloxazolyne derivatives (DMOX) with silver ion-high performance liquid chromatography (Ag+-HPLC). Reconstructed mass spectral profiles of CLA characteristic ions from DMOX were used to identify positional isomers and Ag+-HPLC to quantify them. Total CLA content varied from 0·57 to 0·97 g/100 g of total fatty acids. FAME and DMOX were separated into a complex mixture of minor isomers and major rumenic acid (9-cis 11-trans C18:2) by GC-MS using a 100-m polar capillary column. Rumenic acid would represent more than 75% of total CLA. 11-trans 13-trans, 11–13 cis/trans plus trans/cis and 7–9 cis/trans plus trans/cis were the main CLA isomers after rumenic acid. Minor amounts of 8–10 and 10–12 C18:2 isomers were also found. Although most of the isomers were present in each herd's milk, differences in content were observed for some CLA species.

The composition of the volatile fraction of milk from cows was investigated in a survey of milk samples using a headspace sampling technique and gas chromatography coupled to mass spectrometry analysis (GC-MS). Milk samples were collected from 12 farms, selected for similar management, breed and level of production. Farms were also grouped according to the type of forage in the ration: (1) hay; (2) hay and maize silage; (3) hay, maize silage and grass silages. Forty-one compounds in milk were isolated and identified from GC-MS headspace analysis. Quantitatively, the most representative chemical class was ketones (eight compounds, 170 μg/kg), followed by aldehydes (nine compounds, 63 μg/kg), alcohols (eight compounds, 36 μg/kg), and lower amounts of hydrocarbons (six compounds), sulphur compounds (three compounds), esters (four compounds) and terpenes (three compounds). The novel headspace sampling technique, and the consequent reduction of sample pre-treatment, allowed the identification of low-molecular weight volatile compounds, and reduced the risk of producing artefacts during analysis.

Discriminant analysis was used to identify a classification criterion for milk samples, using type of forage in the ration as a grouping variable. Posterior probability error rate indicated that aldehydes provided one of the best discriminant criteria for grouping milks according to ration composition. When all 41 identified volatile compounds were included, discriminant analysis selected nine compounds (acetone, 2,3-butanedione, 2-butanone, ethanol, acetaldehyde, ethylacetate, ethylisovalerate, dimethylsulphone) that did not fail the tolerance test and which correctly classified 100% of the original cases.