Studies on the Chemical Composition of the Head Space and Essential Oil of Fresh Flower of Ku-Shui rose (Rosa setate X R. rugosa)

The chemical composition of the head space and essential oil of fresh flowers of KU-Shui rose, which were collected by adsorption with Paropark Q (60–80 mesh) and steam distillation-extraction respectively, have been identified both by capillary gas chromatography/mass spectrometry and the Kovats Index of the components. From the head space, 63 compounds have been identified, among which 27 compounds were not present or found only in trace amount in the essential oil. From the essential oil, 79 compounds have been identified, among which 55 compounds have not been found in this flower previously.     

Affinity Selection and Mass Spectrometry-Based Strategies to Identify Lead Compounds in Combinatorial Libraries

The screening of diverse libraries of small molecules created by combinatorial synthetic methods is a recent development which has the potential to accelerate the identification of lead compounds in drug discovery. We have developed a direct and rapid method to identify lead compounds in libraries involving affinity selection and mass spectrometry. In our strategy, the receptor or target molecule of interest is used to isolate the active components from the library physically, followed by direct structural identification of the active compounds bound to the target molecule by mass spectrometry. In a drug design strategy, structurally diverse libraries can be used for the initial identification of lead compounds. Once lead compounds have been identified, libraries containing compounds chemically similar to the lead compound can be generated and used to optimize the binding characteristics. These strategies have also been adopted for more detailed studies of protein–ligand interactions.     

Aliphatic medium chain tricarboxylic acids in rat urine.

Three aliphatic tricarboxylic acids have been found in rat urine. They have been identified as 6-carboxy-5-undecenedioic acid, 6-carboxy-5-dodecenedoic acid, and 6-carboxy-5-tridecenedioic acid. The carbon skeleton structure was determined by mass spectra of the hydrogenated methyl esters. The double bond position was determined after osmium tetroxide oxidation followed by trifluoroacetylation and mass spectrometry and by infrared spectrometry. The compounds were present in the urine when the rats were fed on pellets but disappeared when they received sucrose and water. The acids were not present in the pellets, and a metabolic relation to compounds of longer chain length, possibly mycolic acids, is likely.     

Identification of polycyclic aromatic compounds in mutagenic emissions from steel casting

Workers in ferrous foundries show increased risk of lung cancer. In the steel casting process hot metal is poured into sand moulds solidified with organic binders, producing a plume of smoke containing a variety of organic compounds and showing strong mutagenicity in the Salmonella/S9 assay. We have collected the emissions produced when steel is poured into an experimental sand mould solidified with oil, clay and cereal, a widely used binder system. The organic constituents of these emissions have been fractionated by preparative reverse-phase high performance liquid chromatography (HPLC) and mutagenic fractions have been analysed by capillary column gas chromatography/mass spectrometry (GC/MS). Of the 65 compounds for which mass spectra are reported, 54 have been tentatively identified as alkyl derivatives of polycyclic aromatic compounds. Many compounds of this class are known to be carcinogenic and mutagenic. In addition, several unsubstituted polycyclic aromatic hydrocarbons, including the carcinogenic benz[a]anthracene and benzo[a]pyrene, were found to be present.     

Isolation and Identification of the Constituents of Volatiles of Flowers of Chloranthus spieatus (Tunb.) Makino

The headspace volatiles of the flowers of Chloranthus spicatus (Tunb.) Makino were trapped by a modified circulation adsorption method and were analyzed by GC/MS techniques after desorption by solvent. The componends were identified by mass spectral and temperature-programmed retention index data on dual glass capillary columns (OV-101 and PEG-20 M). 32 compounds have been identified from the volatiles, including 11 monoterpenes, 11 sesquiterpenes, and 7 oxygen-containing compounds.The major consitituents are cis-methyl jasmonatecis-β-ocimene, and β-pinene.     

Study of a virilizing adrenal cortical tumor. Analysis of urinary steroids by gas-liquid chromatography and mass spectrometry]

The C19 and C21 urinary steroids from a virilizing adrenal tumour with high levels of plasma 17alpha-progesterone and its urinary metabolites have been identified and quantitated by gas chromatography and mass spectrometry of sephadex fractions of the total urinary extract. Of the fifty five identified steroids thirteen were new compounds or known compounds not found before in such a case. The actiology of the apparent 21-steroid hydroxylase deficiency is discussed at the light of these analytical results and of the hormonogenesis enzymatic induction of the tumour biopsy.     

Monosaccharides and monosaccharide derivatives in human seminal plasma

Gas chromatography—mass spectrometry with an on-line data system was used to identify monosaccharides and monosaccharide derivatives in human seminal plasma. The carbohydrates were converted into the methoxime—trimethylsilyl derivatives before separation in open tubular glass capillary columns coated with SE-30. Twenty-one different compounds were detected in the seminal fluid, of which twelve have not been recognized before. Seventeen of the monosaccharides have previously been identified in urine. Similar patterns of sugars were found both in fertile and infertile individuals, including one with azoospermia. The compounds identified are, with the possible exception of -ribose, present as free monosaccharides at the time of ejaculation, and they do not seem to be preformed by spermatozoa.     

Identification of ejaculate derived propylamine found in collagen sponge contraceptives

Propylamine is identified as one of the compounds present in odoriferous intravaginal contraceptive sponges from sexually active women. This compound also appears in samples of human ejaculate incubated at 37 degrees C for seven days. We have identified propylamine by gas liquid chromatography, mass spectroscopy and as N-propylbenzoylamide. Evidently the compound forms enzymatically from spermine and spermidine. We believe that this is the first time that propylamine has been identified as forming from human tissue.     

Exploring the Arabidopsis sulfur metabolome

Sulfur plays a crucial role in protein structure and function, redox status and plant biotic stress responses. However, our understanding of sulfur metabolism is limited to identified pathways. In this study, we used a high‐resolution Fourier transform mass spectrometric approach in combination with stable isotope labeling to describe the sulfur metabolome of Arabidopsis thaliana. Databases contain roughly 300 sulfur compounds assigned to Arabidopsis. In comparative analyses, we showed that the overlap of the expected sulfur metabolome and the mass spectrometric data was surprisingly low, and we were able to assign only 37 of the 300 predicted compounds. By contrast, we identified approximately 140 sulfur metabolites that have not been assigned to the databases to date. We used our method to characterize the γ‐glutamyl transferase mutant ggt4‐1, which is involved in the vacuolar breakdown of glutathione conjugates in detoxification reactions. Although xenobiotic substrates are well known, only a few endogenous substrates have been described. Among the specifically altered sulfur‐containing masses in the ggt4‐1 mutant, we characterized one endogenous glutathione conjugate and a number of further candidates for endogenous substrates. The small percentage of predicted compounds and the high proportion of unassigned sulfur compounds identified in this study emphasize the need to re‐evaluate our understanding of the sulfur metabolome.     

N-Coronafacoyl-L-isoleucine and N-coronafacoyl-L-alloisoleucine,potential biosynthetic intermediates of the phytotoxin coronatine

Two new naturally-occurring analogues of the phytotoxin coronatine have been isolated from liquid cultures of Pseudomonas syringae pv. glycinea. These have been identified as N-coronafacoyl-L-isoleucine and N-coronafacoyl-L-alloisoleucine by mass spectrometry and by studies of the products of acid hydrolysis of the two compounds. The compounds were purified as a mixture of ca 2:1 composition, but the two parent components were not preparatively separated. The possible significance of the two compounds, to the biosynthesis of coronatine, is discussed.     

Identification of isomeric dicaffeoylquinic acids from Eleutherococcus senticosus using HPLC-ESI/TOF/MS and 1H-NMR methods

Liquid chromatography-electrospray time-of-flight mass spectrometry (HPLC-ESI/TOF/MS) and a novel NMR technique, developed to maximise the sensitivity obtained from the standard NMR spectrometer, have been applied to the identification of the phenolic constituents of Eleutherococcus senticosus. In addition, molecular modelling and dihedral bond angle calculations based on the vicinal 3JHH-coupling constants have been used in the unambiguous assignment of signals in the 1H-NMR spectra. 5'-O-Caffeoylquinic acid and three isomeric compounds, 1',5'-O-dicaffeoylquinic acid, 3',5'-O-dicaffeoylquinic acid and 4',5'-O-dicaffeoylquinic acid, have been isolated and identified from a sample. The isolation and structure determination of the latter two compounds are reported for the first time from this plant.     

Mass fragmentography. Identification of chlorpromazine and its metabolites in human blood by a new method

Chlorpromazine and some of its metabolites have been identified in human blood with the combination of gas chromatography and mass spectrometry. A new method, mass fragmentography has been elaborated. It is based upon a continuous recording of up to three mass numbers characteristic of a single substance or a group of compounds. With this technique both a high sensitivity and a high selectivity which can be changed according to wish are achieved. Compounds are identified by their retention times and the fact that all mass numbers are represented in characteristic relative intensities. Refocusing on other characteristic fragments and/or the molecular ion confirms the identity. Through repeated refocusing “a partial mass spectrum” of a compound can be established even when the amounts present are too small for the scanning of a complete spectrum.With this method chlorpromazine, and its desmethylated and didesmethylated metabolites have been identified in plasma. The latter two metabolites were obtained also after treatment of the plasma with β-glucuronidase, as was 2-chlorophenothiazinylpropionic acid, which was found in quantities that allowed the scanning of a complete mass spectrum. In red blood cells the two desmethylated metabolites could be identified with mass fragmentography only after treatment with β-glucuronidase. The use of the method is discussed, particularly with regard to blood levels of drugs as related to therapeutic effects and side effects.     

The 8-isoprostaglandins: evidence for eight compounds in human semen.

Evidence is presented for the existence of a group of 8-iso prostaglandins in human semen, comprising 8-iso PG E1, 8-iso PG E2, 8-iso PG F1 alpha, 8-iso PG F2 alpha and the four corresponding 19-hydroxy prostaglandins. The E and F compounds have been positively identified by comparison of their mass spectra and chromatographic properties with those of authentic standards. Preliminary measurements of levels of these compounds in pooled semen are presented.     

Volatile constituents in mosses (Musci)

The essential oils of mosses of the genera Mnium, Plagiomnium, Homalia, Plagiothecium and Taxiphyllum (Musci) have been investigated by gas chromatography and mass spectrometry. The new sesquiterpenes (+)-10-epi-muurola-4,11-diene and 10,11-dihxdro-alpha-cuparenone were isolated by preparative gas chromatography and identified as major constituents of the hydrodistillation products of Mnium hornmum (Hedw.) using NMR and mass spectrometry. In addition, (+)-dauca-8,11-diene and two new butenolides, 3,4,5-trimethyl-5-pentyl-5H-furan-2-one and 3.4-dimethyl-5-pentyl-5H-furan-2-one were identified as constituents in Plagiomnium undulatum (Hedw.) T. Kop. Although the amounts of volatiles present in the investigated mosses are generally smaller than in liverworts, the spectrum of terpenoid compounds is similar. The investigated mosses also generate aliphatic compounds of greater abundance and structural variety.     

Identification of p-hydroxybenzaldehyde as the ligand in the green form of old yellow enzyme.

We have previously reported the isolation of old yellow enzyme complexed with a ligand of low molecular weight which imparts a distinctive charge-transfer absorption to the enzyme, making it green in color (Matthews, R. G., and Massey, V. (1969) J. Biol. Chem. 244, 1779-1789). This ligand has now been identified as p-hydroxybenzaldehyde by removal from the enzyme and characterization of its optical spectrum and subsequent mass spectral analysis. Similar compounds which are also bound to old yellow enzyme have been isolated from yeast extract and identified. These compounds give rise to complexes with old yellow enzyme which are characterized by broad long wavelength absorption bands, and they lead to inhibition of the NADPH-O2 oxidoreductase activity catalyzed by old yellow enzyme.     

Metabolic profiling of Musa acuminata challenged with Sporobolomyces salmonicolor

In vitro plants of Musa acuminata (AAA) “Cavendish” challenged with the non-pathogenic yeast strain Sporobolomyces salmonicolor resulted in the induction of a large number of metabolites. These included phenylphenalenone-type compounds, of which nine have been identified. Two of these are new phenylnaphthalic anhydrides, 2-(4′-β-glucosyloxyphenyl)-1,8-naphthalic anhydride and 2-(3′,4′-dihydroxyphenyl)-1,8-naphthalic anhydride. The chemical structures of these compounds were identified using nuclear magnetic resonance (NMR) spectroscopic and mass spectrometric (MS) techniques.     

2″-p-coumaroylvitexin 7-glucoside from Mollugo oppositifolia

Besides vitexin, two compounds have been isolated from Mollugo oppositifolia and identified as vitexin 7-glucoside and 2″-p-coumaroylvitexin 7-glucoside. The latter is a new natural compound. Some features common to the electron-impact mass spectra of permethyl vitexin 7-glucoside and permethyl isovitexin 7-glucoside are discussed.     

Polar acyclic diterpenoids from Bifurcaria bifurcata (Fucales, Phaeophyta)

From the lipophilic extract of the brown alga Bifurcaria bifurcata collected off the Atlantic coast of Southern Brittany (Quiberon, France), five polar linear diterpenoids have been isolated. These metabolites have been identified as hydroxylated derivatives of 13-oxo- and 13-hydroxygeranylgeraniol. Their structures were characterized on the basis of chemical and spectral evidence including two-dimensional NMR experiments and mass spectrometric techniques. The absolute configuration of the 13-position has been determined, for the 13-hydroxygeranylgeraniol derivatives, to be R by means of a modified Mosher's method and therefore that of 13-hydroxygeranylgeraniol (eleganediol) has been revised. Along with these compounds, three related known geranylgeraniol derivatives were also identified, and these data were used for chemotaxonomical purposes.     

Identification of two novel phytosiderophores secreted by perennial grasses

It has been suggested that some perennial grasses secrete phytosiderophores in response to iron (Fe) deficiency, but the compounds have not been identified. Here, we identified and characterized the phytosiderophores secreted by two perennial grasses, Lolium perenne cv. Tove and Poa pratensis cv. Baron. Root exudates were collected from the roots of Fe-deficient grasses and then purified with various chromatographies. The structure of the purified compounds was determined using both nuclear magnetic resonance and fast atom bombardment mass spectrometry. Both species secreted phytosiderophores in response to Fe deficiency, and the amount of phytosiderophores secreted increased with the development of Fe deficiency. The type of phytosiderophores secreted differed with plant species; L. perenne cv. Tove secreted 3-epihydroxy-2'-deoxymugineic acid (epiHDMA), 2'-deoxymugineic acid (DMA) and an unknown compound, whereas P. pratensis cv. Baron secreted DMA, avenic acid A (AVA) and an unknown compound. Purification and subsequent analysis with nuclear magnetic resonance and mass led to identification of the two novel phytosiderophores; 3-hydroxy-2'-deoxymugineic acid (HDMA) from L. perenne, and 2'-hydroxyavenic acid A (HAVA) from P. pratensis. Both novel phytosiderophores have similar chelating activity to known phytosiderophores.     

Metabolism of the soy isoflavones daidzein, genistein and glycitein in human subjects. Identification of new metabolites having an intact isoflavonoid skeleton

Epidemiological studies have associated high soy intake with a lowered risk for certain hormone-dependent diseases. Soy and soy foods are rich sources of isoflavones, which have been shown to possess several biological activities. In this study, the metabolism of soy isoflavones daidzein, genistein and glycitein was investigated in human subjects. The aim was to find and identify urinary phase I metabolites of isoflavones, which have an intact isoflavonoid skeleton, and which might possess some bioactivity. Six volunteers included three soy bars per day into their normal western diet for a 2-week period. Daily urine samples were collected before, and after the supplementation period. Urine samples were hydrolyzed with Helix pomatia, extracted with diethyl ether, purified with Sephadex LH-20 chromatography, and analyzed as trimethylsilyl derivatives using gas chromatography–mass spectrometry (GC–MS). The structures of the isoflavone metabolites were identified using authentic reference compounds. The metabolites, for which authentic reference compounds were not available, were identified by the interpretation of mass spectra. Several new isoflavone metabolites were identified, and the presence of previously reported metabolites confirmed. The metabolic pathways of daidzein, genistein and glycitein are presented on the basis of the identification of the metabolites in human urine after soy supplementation.