Comparison Of Mass Spectra Research Articles

Two-Dimensional Separation in Laser Ionization Orthogonal Time-of-Flight Mass Spectrometry

The capabilities of two-dimensional separation using a high irradiance laser ionization orthogonal time-of-flight mass spectrometer (LI-O-TOFMS) were demonstrated in this paper. Ions were separated via their initial kinetic energy in one dimension and their mass-to-charge ratios in the other dimension. Investigation of the transient ion profiles after laser pulses revealed that the separation of analyte ions from multiply charged ions and gas species ions was achieved. Comparison of mass spectra in the normal accumulation mode and in the two-dimensional separation mode indicated that the relative sensitivity coefficients are stable and close to their true values in the two-dimensional separation mode, especially for trace elements that are prone to interference.

HS-SPME Comparative Analysis of Genotypic Diversity in the Volatile Fraction and Aroma-Contributing Compounds of Capsicum Fruits from the annuum−chinense−frutescens Complex

Volatile constituents of ripe fruits of 16 Capsicum accessions from the annuum-chinense-frutescens complex, with different aroma impressions and geographical origins, were isolated by headspace-solid phase microextraction (HS-SPME) and analyzed by gas chromatography-olfactometry-mass spectrometry (GC-sniffing port-MS). More than 300 individual compounds could be detected in the studied genotypes; most of them could be identified by comparing mass spectra and retention times with authentic reference substances or literature data. Esters and terpenoids were the main groups, although other minor compounds, such as nitrogen and sulfur compounds, phenol derivatives, norcarotenoids, lipoxygenase derivatives, carbonyls, alcohols, and other hydrocarbons, were also identified. The sniffing test revealed that the diversity of aromas found among the studied cultivars is due to qualitative and quantitative differences of, at least, 23 odor-contributing volatiles (OCVs). C. chinense, and C. frutescens accessions, with fruity/exotic aromas, were characterized by a high contribution of several esters and ionones and a low or nil contribution of green/vegetable OCVs. Different combinations of fruity/exotic and green/vegetable OCVs would explain the range of aroma impressions found among C. annuum accessions. Implications of these findings for breeding and phylogeny studies in Capsicum are also discussed.

Chemical Composition and Mosquito (Aedes aegypti Repellent Activity of Essential Oil Extracted from the Aril of Torreya grandis

This study was performed to provide data on the chemical composition, skin irritant potential and mosquito repellency of Torreya grandis aril essential oil extracted by hydro distillation. Fifty nine compounds were identified through comparison with reference compounds, tentatively identified by comparing mass spectra and Kovats Index with those reported in mass spectra libraries and literature. Monoterpenes and oxygenated monoterpenes represented more than 80 % of the oil. The oil was characterized by high contents of α- pinene (34.20 %), D-limonene (27.58 %), limonene glycol (3.64 %), (E)-carveol (2.50 %) and isoborneol (2.36 %). No positive skin irritant reaction was found within the 15 volunteers exposed to undiluted oil (4-h patch test). Furthermore, the ethanol solutions of the essential oil at several concentrations were tested with arm-in-cage assay for mosquito (Aedes aegypti) repellency. Results showed that the ethanol solutions with essential oil at 12.5 %, 25 %, 50 %, and 100 % (the original undiluted essential oil) provided complete protection of 1.0 ± 0.5, 5.0 ± 0.5, 7.0 ± 1.0, and 7.5 ± 0.5h for human beings, respectively. These results suggest the promising application of the essential oil in natural mosquito repellent production.

A unique proteolytic fragment of alpha1-antitrypsin is elevated in ductal fluid of breast cancer patient

By comparison of mass spectra from a small cohort of nipple aspiration fluids (NAF), we previously discovered a panel of five candidate breast cancer biomarkers among them an unidentified 4.7 kD peptide BF5. The purposes of the present study were to verify the presence of BF5 in an independent cohort; to determine the protein identity of BF5; and to provide insight into the biology of BF5 production and elevation in tumor-associated NAF. We prospectively collected bilaterally matched NAF from patients with unilateral Stage I/II breast cancer (IBC-31), ductal carcinoma in situ (DCIS-6), atypical ductal hyperplasia (ADH-5), and presumed healthy women who came to routine mammography and had a normal exam (31). Following the consolidation of its cancer-associated expression on SELDI-mass spectrometry, BF5 was isolated by gel electrophoresis and sequenced by tandem mass spectrometry. BF5 was elevated in 15-25% of women with IBC, DCIS, or ADH vs. 0% of controls. This elevation was restricted to the affected breasts. BF5 was identified as 41/42-aa C-terminal peptide of alpha1-antitrypsin (AAT), the principle inhibitor of serine protease neutrophile elastase. The full length AAT showed a consistent expression pattern as C-41/42, and C-41/42 can be generated in vitro by MMP-7 cleavage. In conclusion, elevated C-41/42 is likely the result of elevated AAT synthesis, and the activity of specific MMPs present within the tumor. As other C-terminal fragments of AAT are reported to function as tumor-derived suppressors to the host immune-system, elevated C-41/42 may also be predictive of a poor outcome.

Two capa-genes are expressed in the neuroendocrine system of Rhodnius prolixus

CAPA-peptides have been isolated from a broad range of insect species and are typical of the neurosecretory system of the abdominal ventral nerve cord. In Rhodnius prolixus, which is known to transmit Chagas’ disease, CAPA-peptides have potent antidiuretic effects. Recently, two capa-genes were isolated from this hemipteran insect; it was the first description of the occurrence of multiple capa-genes from a single insect species. The expression of peptides from one of these capa-genes was confirmed by mass spectrometry Paluzzi (2008) [10]. In this study, the abdominal neuroendocrine system of R. prolixus was screened for the occurrence of products of the second annotated capa-gene. Single nerve preparations of abdominal segmental nerves 1–3 of R. prolixus were analyzed by MALDI-TOF mass spectrometry and the products of both capa-genes were identified by tandem mass spectrometry. The co-occurrence of the different CAPA-peptides, including CAPA-precursor peptides, in all spectra from abdominal segmental nerves 1 to 3 suggests a common expression of both capa-genes in the same neuroendocrine cells. A comparison of mass spectra obtained from abdominal segmental nerves with those of the subesophageal ganglion (SEG) revealed an obvious differential processing of the CAPA-precursors within the CNS.

Characterization of the volatile fraction of laboratory-generated aerosol particles by thermodenuder-aerosol mass spectrometer coupling experiments

An aerodyne high-resolution time-of-flight aerosol mass spectrometer (HR-ToF-AMS) combined with a thermodenuder (TD) was used to investigate laboratory-generated aerosol particles with regard to their volatility and chemical content. The performance of the setup was tested first by using ammonium sulfate particles. Organic compounds have been measured including humic acid, fulvic acid, succinic acid and its disodium salt, 2-methylsuccinic acid, fumaric acid, α-ketoglutaric acid, and glutaric acid. Among them, humic acid, fulvic acid, and disodium succinate show a non-volatile fraction at 300 °C, while the other organic compounds are more volatile. Comparisons of mass spectra at different temperatures showed that significant differences in mass spectra of humic and fulvic acids are observed, indicating that their molecules changed during or after volatilization. At lower temperatures, the changes in humic acid are more likely due to the evaporation of small organic molecules or decomposition of aliphatic groups. The mass losses of the CO 2 + fragment for both humic and fulvic acids at higher temperatures may arise from decarboxylation processes. The different mass spectra for humic and fulvic acids before and after TD also suggest that one should be careful in interpretation of volatility measurements because some molecule structures may change after being heated, especially for the non-volatile multifunctional compounds.

Capabilities of Direct Sample Introduction−Comprehensive Two-Dimensional Gas Chromatography−Time-of-Flight Mass Spectrometry to Analyze Organic Chemicals of Interest in Fish Oils

Most analytical methods for persistent organic pollutants (POPs) focus on individual groups of targeted analytes. Therefore, analysis of multiple classes of POPs typically entails several sample preparations, fractionations, and injections, whereas other chemicals of possible interest are neglected or lost. To analyze a wider scope of organic contaminants in fish oil, we developed an approach to combine the analysis of targeted and untargeted chemicals using an automated direct sample introduction (DSI) and comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (GC x GC/ ToF-MS). DSI-GC x GC/ToF-MS is a powerful tool that attains high quality separations to achieve high selectivity while still providing a wide analytical scope with minimal sample preparation, especially in conjunction with DSr's high tolerance to dirty extracts. Gel permeation chromatography (GPC) was used for initial separation of lipids from POPs and other GC-amenable organic compounds from dietary cod liver oil. For comparison purposes, additional cleanup of the GPC extracts was done by silica adsorption and acidification, which helped provide clues in the identification of untargeted compounds, but in routine analysis, only GPC is needed for this analytical approach. The approach allowed simultaneous identification of known-POPs in the fish liver oils, and further permitted presumptive identifications of multiple groups of halogenated natural products (HNPs) and other organic chemicals of interest through comparisons of the mass spectra from analyses with those from mass spectral libraries and/or reports in the literature (approximately 60 PCB congeners and 76 compounds in total). Subsequent confirmations were made by reanalysis and comparison of chromatographic retention times and mass spectra with contemporaneously analyzed reference standards. Otherwise, ion fragmentation patterns of unknown compounds were assessed for tentative identifications. Some of the HNPs in the fish oils were detected and identified for the first time. Our study demonstrates that the wide monitoring scope provided by the DSI-GC x GC/ToF-MS method after GPC provides many logistical and performance advantages over the conventional use of several different methods designed for individual classes of targeted analytes after extensive sample preparation.

Profile of Volatile Compounds in 11 Brandies by Headspace Solid‐Phase Microextraction Followed by Gas Chromatography‐Mass Spectrometry

A headspace solid-phase microextraction (HS-SPME) method coupled with gas chromatography-mass spectrometry (GC-MS) was applied for the qualitative or semiquantitative characterization of brandy volatiles. SPME variables (SPME fiber, extraction temperature and time, and ethanol concentration) were optimized. A total of 144 compounds were from the brandies' volatiles, tentatively identified or identified by comparing mass spectra and retention indices of the standards or from literature. Of these, 57 are common to 11 brandies. They were mainly represented by esters and alcohols, such as 2-methyl propanol, 3-methyl butanol, 1-hexanol, ethyl octanoate, and ethyl decanoate, which were quantitatively determined. Chromatographic peaks were integrated using selective ion method (SIM) and the semiquantitative data analyzed using principal component analysis (PCA) and cluster analysis (CA) to study relationships between volatile composition and brandy. Eleven brandies were differentiated into 3 groups: 1 for Hennessy VSOP and XO samples, 1 for Changyu PEGASE VSOP and XO-1, 2, 3 samples, and the other for Changyu PEGASE brandy and VO, Taro brandy, Baiyang River brandy, and Wealth XO samples. The classification of groups is consistent with the brandy samples by variety and grade.

Response on “Regulations in the field of residue and doping analysis should ensure a well-defined risk of a false positive declaration” by N. M. Faber

Dr. Faber provided support for a valuable methodology for the comparison of mass spectra. However, since many other factors are involved in the evaluation process before declaring a sample “positive”, i.e. reporting an adverse analytical finding, his conclusions on this subject are incorrect. In this paper a number of arguments are listed showing that Dr. Faber did not take all factors under consideration when making his claims. These factors involve parameters directly related to mass spectrometric criteria, but also to other equally important factors including sample preparation specificity, retention time (since in doping control mass spectrometry is used as part of a hyphenated technique), etc. These arguments clearly illustrate that the risks for a false positive declaration can not be calculated according to Dr. Faber’s proposed methodology.

Biological activity and identification of neuropeptides in the neurosecretory complexes of the cabbage pest insect,Mamestra brassicae(Noctuidae; Lepidoptera)

The need for more environmentally sound strategies of plant protection has become a driving force in physiological entomology to combat insect pests more efficiently. Since neuropeptides regulate key biological processes, these "special agents" or their synthetic analogues, mimetics, agonists or antagonists may be useful tools. We examined brain-suboesophageal ganglia and corpora cardiaca-corpora allata complexes of the cabbage moth, Mamestra brassicae, in order to obtain clues about possible peptide candidates which may be appropriate for the biological control of this pest. With the aid of bioassays, reversed phase high performance liquid chromatography, and mass spectrometry, five neuropeptides were unequivocally identified and the presence of a further three were inferred solely by comparing mass spectra with known peptides. Only one neuropeptide with adipokinetic capability was identified in M. brassicae. Data from the established homologous bioassay indicated that the cabbage moths rely on a lipid-based metabolism which is aided by an adipokinetic hormone (viz. Manse-AKH) that had previously been isolated in many different lepidopterans. Other groups of neuropeptides identified in this study are: FLRFamides, corazonin, allatostatin and pheromonotropic peptide.

Rapid‐resolution liquid chromatography/mass spectrometry for determination and quantitation of polyphenols in grape berries

A rapid-resolution liquid chromatography/mass spectrometric (RRLC/MS) method for detection and quantitation of polyphenols in grape berry skins and seeds has been developed. Pulp-free berry skins were treated with liquid nitrogen and ground; seeds were also ground. Then, 3 g of samples were extracted with 30 mL of a mixture of methanol/water/formic acid 70:30:1 (v/v/v) under sonication and 1 microL of the final extract was injected into two 100 x 2.1 mm i.d., 1.8 microm Zorbax Eclipse plus C18 columns connected in series. Compounds were fractionated using a gradient elution of acidified acetonitrile/methanol 50:50 (v/v)/water. Columns were thermostatted at 70 degrees C. MS was carried out on an Agilent 6410 QqQ instrument equipped with an electrospray ionization source. Positive and negative MS/MS product ion scans were used for compound identification, whereas positive full scan MS in the m/z range 200-1400 was used for quantitation. By means of mass spectra comparison, various flavonols, flavan-3-ols, anthocyanins and stilbenes were identified. Quantitation was performed by external calibration, and concentration values were corrected for matrix effect that was evaluated in separate experiments. Semi-quantitative estimation was performed for compounds for which standards were not commercially available. Recoveries ranged from 90-102% with relative standard deviation (RSD) <5%, whereas the between samples RSD was in the range 4-12%. Two surrogate standards were used for quality control. The developed method was applied to analyze the polyphenol content of three Vitis vinifera table cultivars at physiological maturity and after proper preservation for 6 weeks. Results demonstrated that during preservation about half of the polyphenol content was lost.

Comparison of mass spectra of peptides in different matrices using matrix‐assisted laser desorption/ionization and a multi‐turn time‐of‐flight mass spectrometer, MULTUM‐IMG

The mass spectra of peptides obtained with different matrices were compared using a matrix-assisted laser desorption/ionization (MALDI) ion source and a multi-turn time-of-flight (TOF) mass spectrometer, MULTUM-IMG, which has been developed at Osaka University. Two types of solid matrices, alpha-cyano-4-hydroxycinnamic acid (CHCA) and 2,5-dihydroxybenzoic acid (DHB), and a liquid matrix made from a mixture of 3-aminoquinoline and CHCA were used. When measuring the peak signal intensity of human angiotensin II [M+H]+ from a fixed sample position, the liquid matrix produced a stable signal over 1000 laser shots, while the signal obtained with CHCA and DHB decayed after about 300 and 100 shots, respectively. Significant differences in the mass resolving power were not observed between the spectra obtained with the three matrices. Signal peak areas were measured as a function of the cycle number in a multi-turn ion trajectory, i.e., the total flight time over a millisecond time scale. For both [M+H]+ of human angiotensin II and bovine insulin, the decay of the signal peak area was the most significant with CHCA, while that measured with DHB was the smallest. The results of the mean initial ion velocity measurements suggested that the extent of metastable decomposition of the analyte ions increased in order of DHB, the liquid matrix, and CHCA, which is consistent with the difference in the decay of the signal peak area as the total flight time increased.

Determination of Oleanolic, Betulinic and Ursolic Acid in Lamiaceae and Mass Spectral Fragmentation of Their Trimethylsilylated Derivatives

Phytochemical investigations of different species of Lamiaceae family (rosemary-Rosmarinus officinalis L., sage-Salvia officinalis L., winter savory-Satureja montana L., clary sage-Salvia sclarea L. and sticky sage-Salvia glutinosa L.), using gas chromatography and mass spectrometry (GC-MS) were performed. The studies were focused on oleanolic, betulinic and ursolic acid. Since oleanolic and ursolic acid are position isomers with very similar structures, the difficulties in their separation and identification have been reported by several authors. However, both compounds can be well distinguished by order of elution during gas chromatography and by intensities of the fragment ion signals in their mass spectra, where the retro-Diels-Alder reaction was primarily observed. A combination of ultrasonic extraction, solid phase extraction, size exclusion chromatography, trimethylsilylation, and GC-MS resulted in simultaneous separation, identification and quantification of the mentioned compounds. The compounds have been identified by retention time and comparison of mass spectra with those of pure standards. The mass spectral fragmentation behavior of all three derivatised acids was investigated. The obtained characteristic fragment patterns are discussed in the presented work. Good linearity over the concentration range 1–50 mg L−1 for all three compounds was confirmed. The correlation coefficients (r 2 ) were in the range of 0.9980–0.9983. Quantitative analyses of different Lamiaceae extracts showed that the oleanolic acid content ranged from 0.09 to 0.9% dry weight, content of betulinic acid ranged from traces to 0.6%, and that of ursolic acid varied from 0.09 to 1.6% dry weight.

Detection of 2-thiaadamantanes in the oil from Well TZ-83 in Tarim Basin and its geological implication

Thermochemical sulfate reduction (TSR) causes increasing of sulfur content in the oils, reduction of oil quality, and produces a toxic and corrosive H2S which may take great adverse effect on health and equipment. Although there are several ways to identify presence of TSR in gas field, yet in the case of oil with low H2S content, it is still very difficult to identify TSR. 2-thiaadamantanes is one of the most effective indicators of TSR. This paper for the first time discovers 2-thiaadamantanes in the oil from Well TZ-83 in Tarim Basin. A silver nitrate silica gel chromatography was used to enrich the oil from low 2-thiaadamantanes content to the level that GC/MS and GC/MS/MS can detect. 2-thiaadamantanes were assigned by comparison of mass spectra with published data, and the fragmental ions in mass spectra were confirmed by MRM mode of GC/MS/MS. H2S gas produced in Tazhong area is thought to be generated from TSR since 2-thiaadamantanes were present in the oil.

Analysis of polycyclic aromatic hydrocarbon sequences in a premixed laminar flame by on‐line time‐of‐flight mass spectrometry

A time-of-flight mass spectrometer in reflectron configuration has been used for the real-time detection of combustion products. The products of a premixed laminar C2H4/O2 flame at atmospheric pressure were sampled along its axis, diluted with inert gas and carried to the ion source as a molecular beam under minimal perturbation. Electron ionization and different optical ionization sources are compared. Photoionization was achieved with laser radiation from a Nd:YAG nanosecond pulsed laser at two different wavelengths in the UV range (266 and 355 nm). The mass spectra obtained using laser wavelength of 355 nm and electron ionization present a series of ions regularly spaced by 18 m/z units up to m/z 2000. This series allowed precise calibration of the instrument for compounds of high molecular weight. Information on the chemical nature of the analyzed species has been obtained by comparing mass spectra produced with different ionization methods. In order to better understand the growth mechanisms, polycyclic aromatic hydrocarbon sequences have been analyzed by fast Fourier transform of the mass spectra.

Analysis of the leaf Oil of Syzygium malaccense Merr. et Perry from Nigeria

The hydrodistilled essential oil from fresh leaves of Syzygium malaccense Merr. et Perry. grown in Nigeria has been studied by GC and GC/MS for the first time. Identification of the substances was made by comparison of mass spectra and retention indices with literature records. The oil was largely composed of monoterpenes (61.1%) characterized mainly by (+)-α-pinene (7.3%), (−)-β-pinene (8.0%), p-cymene (13.5%), and α-terpineol (7.5%). The sesquiterpenes constituted 30.8% of the oil with (−)-β-caryophyllene (9.0%) as the major component.

Identification of Glioma Neovascularization-related Proteins by Using MALDI-FTMS and Nano-LC Fractionation to Microdissected Tumor Vessels

The identification of angiogenesis-related proteins is important for the development of new antiangiogenic therapies, and such proteins are potential new biomarkers for gliomas. The aim of this study was to identify proteins that are exclusively present in glioma neovasculature and not in the vasculature of normal brain. We combined advanced proteomics techniques to compare the expression profiles of microdissected blood vessels from glioma with blood vessels of normal control brain samples. We measured the enzymatic generated peptide profiles from these microdissected samples by MALDI-FTMS. Subsequently, the samples were fractionated by nano-LC prior to MALDI-TOF/TOF. This combined approach enabled us to identify four proteins that appeared to be exclusively expressed in the glioma blood vessels. Two of these proteins, fibronectin and colligin 2, were validated on tissue sections using specific antibodies. We found that both proteins are present in active angiogenesis in glioma, other neoplasms, and reactive conditions in which neoangiogenesis takes place. This work proves that gel-free mass spectrometric techniques can be used on relatively small numbers of cells generated by microdissection procedures to successfully identify differentially expressed proteins.

SAMPI: protein identification with mass spectra alignments.

BackgroundMass spectrometry based peptide mass fingerprints (PMFs) offer a fast, efficient, and robust method for protein identification. A protein is digested (usually by trypsin) and its mass spectrum is compared to simulated spectra for protein sequences in a database. However, existing tools for analyzing PMFs often suffer from missing or heuristic analysis of the significance of search results and insufficient handling of missing and additional peaks.ResultsWe present an unified framework for analyzing Peptide Mass Fingerprints that offers a number of advantages over existing methods: First, comparison of mass spectra is based on a scoring function that can be custom-designed for certain applications and explicitly takes missing and additional peaks into account. The method is able to simulate almost every additive scoring scheme. Second, we present an efficient deterministic method for assessing the significance of a protein hit, independent of the underlying scoring function and sequence database. We prove the applicability of our approach using biological mass spectrometry data and compare our results to the standard software Mascot.ConclusionThe proposed framework for analyzing Peptide Mass Fingerprints shows performance comparable to Mascot on small peak lists. Introducing more noise peaks, we are able to keep identification rates at a similar level by using the flexibility introduced by scoring schemes.

Direct detection and quantification of 19-norandrosterone sulfate in human urine by liquid chromatography-linear ion trap mass spectrometry

19-Norandrosterone sulfate (19-NAS) is the sulfoconjugated form of 19-norandrosterone (19-NA), the major metabolite of the steroid nandrolone. A sensitive and accurate liquid chromatography/tandem mass spectrometry (LC-MS/MS) assay was developed for the direct measurement of 19-NAS in human urine samples. The method involved a quaternary amine SPE protocol and subsequently injection of the extract onto an analytical column (Uptisphere ® ODB, 150 mm × 3.0 mm, 5 μm) for chromatographic separation and mass spectrometry detection in negative electrospray ionisation mode. The sulfoconjugate of 19-NA was identified in urine by comparison of mass spectra and retention time with a reference substance. The limit of detection (LOD) and lowest limit of quantification (LLOQ) of 19-NAS were of 40 pg/mL and 200 pg/mL, respectively. For a nominal concentration of 2 ng/mL, recovery (94%), intra-day precision (2.7%), intra-assay precision (6.6%) and inter-assay precision (14.3%) were determined. Finally, this analytical method was applied for quantifying the concentration of 19-NAS in doping samples, using calibration curves (0.2–20 ng/mL) and the standard-addition method. The results show the feasibility of applying this LC-MS/MS assay as a complementary tool to detect misuse of nandrolone or nandrolone precursors.

Composition and antimicrobial activity of the essential oil of six Hypericum species from Serbia

The volatile composition of six Hypericum species has been studied. The essential oils were obtained by steam distillation in 500 mL H 2O for 2 h in a modified Clevenger apparatus with a water-cooled oil receiver to reduce hydrodistillation over-heating artifacts, and their analyses were performed by GC and GC–MS. Identification of the substances was made by comparison of mass spectra and retention indices with literature records. A total of 100 different compounds were identified. The main constituents of the investigated populations of each taxon have been revealed as follows: Hypericum alpinum: (−)-β-pinene, γ-terpinene, (−)-( E)-caryophyllene; Hypericum barbatum: (−)-α-pinene, (−)-β-pinene, (−)-limonene, (−)-( E)-caryophyllene, (−)-caryophyllene oxide; Hypericum rumeliacum: (−)-α-pinene, (−)-β-pinene, (−)-limonene, Hypericum hirsutum: nonane, undecane, (−)-( E)-caryophyllene, (−)-caryophyllene oxide; Hypericum maculatum: spathulenol, globulol; Hypericum perforatum: (−)-α-pinene, ( Z)-β-farnesene, germacrene D; Monoterpene hydrocarbons were shown to be the main group of the taxa belonging to the section Drosocarpium, while the taxa of section Hypericum were more rich in sesquiterpene hydrocarbons.