Hernandulcin Production in Elicited Hairy Roots of Phyla scaberrima: Toward Sustainable Production of a Non-Caloric Sweetener with Nutraceutical Properties.

Herein we report on the generation of hairy root lines of P. scaberrima able to produce hernandulcin (HE), a non-caloric sweetener with nutraceutical properties. From ten different lines analyzed, three synthesized up to 100 mg ⋅ L-1 HE under the batch culture conditions standardized in this investigation. Adding elicitors (salicylic acid, chitin, Glucanex, polyethylene glycol) and biosynthetic precursors (farnesol and (+)-epi-alpha-bisabolol) significantly altered HE accumulation. Chitin and Glucanex enhanced HE production from 130 to 160 mg ⋅ L-1 , whereas farnesol and (+)-epi-alpha-bisabolol from 165 to 200 mg ⋅ L-1 without dependence on biomass accumulation. Improved batch cultures containing liquid Murashige & Skoog medium (MS; pH 7), added with 4 % sucrose, 0.5 mg ⋅ L-1 naphthaleneacetic acid, 100 mg ⋅ L-1 Glucanex, 150 mg ⋅ L-1 chitin, 250 mg ⋅ L-1 farnesol, and 150 mg ⋅ L-1 (+)-epi-alpha-bisabolol at 25 °C (12 h light/12 h darkness), triggered HE accumulation to 250 mg ⋅ L-1 in 25 days. The efficiency of each recombinant line is discussed.

Unambiguous determination of farnesol and tyrosol in vaginal fluid using fast and sensitive UHPLC-MS/MS method.

The new ultra-high performance liquid chromatography method with tandem mass spectrometry detection (UHPLC-MS/MS) has been optimized to allow fast, selective, and high-throughput analysis of two Candida albicans quorum sensing molecules (QSM), farnesol and tyrosol. The problem of the presence of the interference in the samples and system was successfully solved by careful optimization of chromatographic conditions. Charged hybrid stationary phase modified with pentafluorophenyl group and optimized gradient elution provided adequate separation selectivity and peak shapes. The impurity was identified as dibutyl phthalate and had the same m/z ions as farnesol leading to an important interference on selected reaction monitoring channel. Two different types of biological matrices originating from vaginal fluid, supernatant and sediment, were analysed. Micro-solid phase extraction in pipette tips was optimized for the selective isolation of QSM from the supernatant. The insufficient retention of farnesol on the extraction sorbent was improved when 1% of organic solvent was added prior to extraction, while the retention of tyrosol was only possible when using combined C8 and polymer sorbent type. Strong retention of farnesol had to be solved by increasing elution solvent strength and volume up to 600 µL. However, this approach did not allow the pretreatment of sediment samples due to the sorbent clogging. Therefore, our previously developed protein precipitation method was modified and validated to analyse the sediments. New developed UHPLC-MS/MS method provided suitable accuracy and precision for the determination of QSM in vaginal fluid while using only 50 µL sample volume and two different sample preparation methods.

A rapid quantitative assay for juvenile hormones and intermediates in the biosynthetic pathway using gas chromatography tandem mass spectrometry.

A method for rapid quantitation of insect juvenile hormones (JH) and intermediates in the biosynthetic pathway, both in vitro and in vivo (hemolymph and whole body), has been developed using GC-MS/MS. This method is as simple as the radiochemical assay (RCA), the most commonly used method for measurement of JH biosynthesis in vitro, without need for further purification and derivatization, or radioactive precursors or ligands. It shows high sensitivity, accuracy and reproducibility. Linear responses were obtained the range of 1-800 ng/mL (approximately 4-3000 nM). Recovery efficiencies for farnesol, farnesal, methyl farnesoate and JH III were approximately 100% in vitro and over 90% in vivo, with excellent reproducibility at three different spike levels. Titer of JH III in the hemolymph was relatively low at day 0 (adult female emergence) (79.68 ±â€¯5.03 ng/mL) but increased to a maximum of 1717 ng/mL five days later. In whole body, JH III quantity reached a maximum on day 4 (845.5 ±â€¯87.9 ng/g) and day 5 (679.7 ±â€¯164.6 ng/g) and declined rapidly thereafter. It is in agreement with the hemolymph titer changes and biosynthetic rate of JH in vitro. Comparison with the results of inhibition of JH biosynthesis by two known inhibitors (allatostatin (AST) mimic H17 and pitavastatin) using RCA and GC-MS/MS, showed that there was little difference between the two methods In contrast to other methods, the present method with GC-MS/MS can be used to elucidate the mechanism of inhibition by inhibitors of JH biosynthesis without any derivatization and purification. This method is applicable to screening of JH inhibitors and the study of inhibitory mechanisms with high sensitivity and accurate quantification. It may also be useful for the determination of JH titer in other Arthropods.

Analysis of essential oils and fragrances with a new generation of highly inert gas chromatographic columns coated with ionic liquids.

In the fields of essential oils and fragrances, samples often consist of mixtures of compounds with similar structural and physical characteristics (e.g. mono- and sesquiterpenoids), whose correct identification closely depends on the synergic combination of chromatographic and mass spectral data. This sample complexity means that new GC stationary phases with different selectivities are continually being investigated. Ionic liquids (ILs) are of great interest as GC stationary phases in this field because of their selectivity (significantly different than that of currently phases) and their high temperature stability. A first generation of IL GC columns was found to be competitive when applied to these field, in terms of selectivity and efficiency, compared to conventional columns (polydimethylsiloxane, (e.g. OV-1), methyl-polysiloxane 5%-phenyl (e.g. SE-52), 7%-cyanopropyl, 7%-phenyl polysiloxane (e.g. OV-1701), and polyethylen glycol (e.g. PEG-20M). However, these columns showed significant activity towards polar or active analytes, which primarily affected their quantitative performance. A new generation of highly-inactive columns coated with three of the most widely-used ionic liquid GC stationary phases has recently been introduced; these phases are SLB-IL60i (1,12-di(tripropylphosphonium) dodecane bis(trifluoromethylsulfonyl) imide [NTf2], SLB-IL76i (tri-(tripropylphosphonium-hexanamido)-triethylamine [NTf2]), and SLB-IL111i (1,5-di (2,3-dimethyllimidazolium) pentane [NTf2]). This study carefully tested the new inert IL columns, in view of their routine application in the fragrance and essential oil fields. They were found to have unusually high selectivity, comparable to that of first-generation IL columns, while their inertness and efficiency were competitive with those of currently-used conventional columns. The IL column performance of first and second generations was compared, through the quali-quantitative analysis of components in a group of different complexity samples; these included the Grob test, a standard mixture of "suspected" skin allergens, and the essential oils of chamomile and sandalwood.

[Effect of andrographolide on quorum sensing and relevant virulence genes of Candida albicans].

OBJECTIVE: To investigate the effect of andrographolide (AG) on quroum sensing (QS) and relevant virulence genes of Candida albicans. METHOD: Gas-chromatography-mass spectrometry (GC-MS) was applied to detect the changes in the content of farnesol and tyrosol in C. albicans intervened by AG. The real-time quantitative PCR (qRT-PCR) was adopted to inspect the expressions of relevant virulence genes such as CHK1, PBS2 and HOG1 regulated by QS. RESULT: At 2 h after the growth of C. albican, the farnesol and tyrosol secretions reduced, without notable change after intervention with AG. The secretions were highest at 12 h and decreased at 24 h. After the intervention with different concentrations of AG, the farnesol content reduces, whereas tyrosol increased, indicating a dose-dependence, particularly with 1 000 mg x L(-1) AG. qRT-PCR revealed that 1 000 mg x L(-1) AG could down-regulate CHK1 by 2.375, 3.330 and 4.043 times and PBS2 by 2.010, 4.210 and 4.760 times, with no significant change in HOG1. CONCLUSION: AG could inhibit the farnesol secretion, promote the tyrosol secretion and down-regulate QS-related virulence genes CHK1 and PBS2 expressions.

Chemical features of Pericarpium Citri Reticulatae and Pericarpium Citri Reticulatae Viride revealed by GC-MS metabolomics analysis.

This paper introduces a detailed method to apply metabolic profiles conducting on tangerine peels (Citrus reticulata 'Dahongpao') at three maturity stages from July to December. Principal component analysis not only demonstrated the metabolic footprints of tangerine peels during ripening but also revealed the compounds (D-limonene and linalool) that mostly contributed to it. Furthermore, some other characteristic compounds were screened to further reveal the chemical features of Pericarpium Citri Reticulatae (PCR) and Pericarpium Citri Reticulatae Viride (PCRV). In particular, compounds such as 4-carene (r = -0.94), 3-carene (r = -0.91), ß-pinene (r = -0.85) and γ-terpinene (r = -0.87) were screened as major components for the pungent smell of PCRV. Geranyl acetate (r = 0.81), farnesyl acetate (r = 0.87) and three alcohols (6-hepten-1-ol, 3-methyl-1-hexanol, 1-octanol) may lead to the pleasant odour of PCR. We therefore propose that the metabolomics analysis focusing on ripening process will be an effective strategy for quality control of closely related herbal medicines.

European Directive fragrances in natural products.

Information on the presence of European Directive fragrance (EUF) allergens in plants and foods is important for numerous reasons. If an individual is allergic to an EUF and is avoiding fragrance, it is possible that they may still be exposed to the allergen in a natural product. In addition, because many of these allergens are also found in foods, it is possible that ingestion of a food containing the allergen may induce systemic contact allergy. Finally, individuals with lip dermatitis may react to contact with foods that contain the allergen. In this article, we have used the data available to identify which plants and foods contain EUF. When available, concentrations of EUF in natural products are provided. The goal of this article is to narrow down the list of botanicals to avoid for specific EUF allergies.

Comparative study of the chemical composition and biological activities of Magnolia grandiflora and Magnolia virginiana flower essential oils.

The biological activities and the determined major volatile components in the Magnolia grandiflora and M. virginiana flowers extracts were compared. Volatile components were detected in the essential oil by dynamic headspace sampling (HS). 2-Phenylethanol (40% and 61%) was found as the main constituent in the essential oil and HS samples of M. virginiana, respectively. In the M. grandiflora oil sample, (E,E)-farnesol (18%) and 2-phenylethanol (10%) were found as main constituents, whereas germacrene D (17%) and ß-bisabolene (17%) were the main components of the HS sample. The essential oil in M. virginiana displayed a moderate antioxidant activity relative to vitamin E, whereas both essential oils were active against human lung carcinoma and breast carcinoma cell lines, even at concentrations higher than 200 µg mL(-1).

Identification, functional characterization, and regulation of the enzyme responsible for floral (E)-nerolidol biosynthesis in kiwifruit (Actinidia chinensis).

Flowers of the kiwifruit species Actinidia chinensis produce a mixture of sesquiterpenes derived from farnesyl diphosphate (FDP) and monoterpenes derived from geranyl diphosphate (GDP). The tertiary sesquiterpene alcohol (E)-nerolidol was the major emitted volatile detected by headspace analysis. Contrastingly, in solvent extracts of the flowers, unusually high amounts of (E,E)-farnesol were observed, as well as lesser amounts of (E)-nerolidol, various farnesol and farnesal isomers, and linalool. Using a genomics-based approach, a single gene (AcNES1) was identified in an A. chinensis expressed sequence tag library that had significant homology to known floral terpene synthase enzymes. In vitro characterization of recombinant AcNES1 revealed it was an enzyme that could catalyse the conversion of FDP and GDP to the respective (E)-nerolidol and linalool terpene alcohols. Enantiomeric analysis of both AcNES1 products in vitro and floral terpenes in planta showed that (S)-(E)-nerolidol was the predominant enantiomer. Real-time PCR analysis indicated peak expression of AcNES1 correlated with peak (E)-nerolidol, but not linalool accumulation in flowers. This result, together with subcellular protein localization to the cytoplasm, indicated that AcNES1 was acting as a (S)-(E)-nerolidol synthase in A. chinensis flowers. The synthesis of high (E,E)-farnesol levels appears to compete for the available pool of FDP utilized by AcNES1 for sesquiterpene biosynthesis and hence strongly influences the accumulation and emission of (E)-nerolidol in A. chinensis flowers.

Chemical composition and antifungal activity of Matricaria recutita flower essential oil against medically important dermatophytes and soil-borne pathogens.

OBJECTIVE: Fungal infections are potential public health threats all over the world. In the present study, effect of Matricaria recutita flower essential oil (EO) was evaluated against medically important dermatophytes and opportunistic saprophytes using microbioassay technique. MATERIALS AND METHODS: Flower essential oil (EO) of M. recutita prepared by hydrodistillation was analyzed by gas chromatography/mass spectrometry (GC/MS). The effect of plant EO on the growth of pathogenic dermatophytes and opportunistic saprophytes was assessed using microbioassay technique. In the bioassay, fungi were cultured in 6-well flat-bottom microplates in presence of various concentrations of plant EO (2.5-1000µg/mL) for 4-10days at 28°C. RESULTS: A total of 14 compounds were identified in the plant oil by GC/MS accounting for 97.5% of the oil composition. The main compound identified was chamazulene (61.3%) followed by isopropyl hexadecanoate (12.7%), trans-trans-farnesol (6.9%) and E-ß-farnesol (5.2%). Growth inhibition for the dermatophytes exposed to serial two-fold concentrations of plant EO (2.5 to 80µg/mL) was reported in the range of 3.24 to 68.15% for Microsporum gypseum, 24.48 to 100% for M. canis, 11.40 to 96.65% for Trichophyton mentagrophytes, 27.79 to 100% for T. rubrum and 45.73 to 100% for T. tonsurans. M. recutita EO inhibited the growth of opportunistic saprophytes by 3.98 to 64.29% for Aspergillus flavus, 6.38 to 93.62% for A. fumigatus, 3.52 to 89.45% for A. niger, 6.38 to 77.66% for Trichoderma harzianum and 17.41 to 89.41% for Fusarium oxysporum in serial two-fold concentrations of 15.62 to 1000µg/mL. CONCLUSION: Results of the present study indicate that M. recutita could be considered as a potential candidate for designing effective antifungal formulations suitable for treatment of dermatophytosis and other fungal infections.

Zap1 control of cell-cell signaling in Candida albicans biofilms.

Biofilms of Candida albicans include both yeast cells and hyphae. Prior studies indicated that a zap1Δ/Δ mutant, defective in zinc regulator Zap1, has increased accumulation of yeast cells in biofilms. This altered yeast-hypha balance may arise from internal regulatory alterations or from an effect on the production of diffusible quorum-sensing (QS) molecules. Here, we develop biosensor reporter strains that express yeast-specific YWP1-RFP or hypha-specific HWP1-RFP, along with a constitutive TDH3-GFP normalization standard. Seeding these biosensor strains into biofilms allows a biological activity assay of the surrounding biofilm milieu. A zap1Δ/Δ biofilm induces the yeast-specific YWP1-RFP reporter in a wild-type biosensor strain, as determined by both quantitative reverse transcription-PCR (qRT-PCR) gene expression measurements and confocal microscopy. Remediation of the zap1Δ/Δ zinc uptake defect through zinc transporter gene ZRT2 overexpression reverses induction of the yeast-specific YWP1-RFP reporter. Gas chromatography-mass spectrometry (GC-MS) measurements of known organic QS molecules show that the zap1Δ/Δ mutant accumulates significantly less farnesol than wild-type or complemented strains and that ZRT2 overexpression does not affect farnesol accumulation. Farnesol is a well-characterized inhibitor of hypha formation; hence, a reduction in farnesol levels in zap1Δ/Δ biofilms is unexpected. Our findings argue that a Zap1- and zinc-dependent signal affects the yeast-hypha balance and that it is operative in the low-farnesol environment of the zap1Δ/Δ biofilm. In addition, our results indicate that Zap1 is a positive regulator of farnesol accumulation.

[Analysis on the compositions of volatile oil of Alpinia henryi with GC-MS method].

OBJECTIVE: To study the components of volatile oil of Alpinia henryi. METHODS: The volatile oil was extracted by steam distillation method,used gaseous phase-mass spectrum combination method (GC-MS) to analyze the components of volatile oil. RESULTS: 58 kinds of components were isolated, among them 42 were identified and determined the relative content. CONCLUSION: This study provides a basis for the development and utilization of Alpinia henryi.

2,3-Dihydrohomofarnesal: female sex attractant pheromone component of Callosobruchus rhodesianus (Pic).

Callosobruchus rhodesianus (Pic) (Coleoptera: Chrysomelidae: Bruchinae) is a pest of stored legumes through the Afro-tropical region. In laboratory bioassays, males of C. rhodesianus were attracted to volatiles collected from virgin females. Collections were purified by various chromatographic techniques, and the biologically active component isolated using gas chromatographic-electroantennographic detection analysis. Gas chromatography-mass spectrometry and NMR analyses suggested that the active compound was 2,3-dihydrohomofarnesal, i.e., 7-ethyl-3,11-dimethyl-6,10-dodecadienal. The structure was confirmed by non-stereoselective and enantioselective total synthesis. Using chiral gas chromatography, the absolute configuration of the natural compound was confirmed as (3S,6E)-7-ethyl-3,11-dimethyl-6,10-dodecadienal. Y-tube olfactomter assays showed that only the (S)-enantiomer attracted males of C. rhodesianus. The (R)-enantiomer and racemate did not attract males, suggesting that the (R)-enantiomer inhibits the activity of the natural compound. In combination with previous reports about sex attractant pheromones of congeners, we suggest that a saltational shift of the pheromone structure arose within the genus Callosobruchus.

Ultra high performance liquid chromatography tandem mass spectrometry analysis of quorum-sensing molecules of Candida albicans.

Candida albicans is generally one of the most commonly isolated fungal pathogen from human body. It is a frequent cause of nosocomial infections, bloodstream infections, urinary infections and mucosal infections of oral cavity and vagina C. albicans can grow as hyphae, pseudohyphae, or budding yeast. Morphological conversion of a yeast form to pseudohyphal or hyphal one is often characterized by the change of commensal status to an invasive form. Farnesol and tyrosol can participate in these transformation processes as quorum sensing molecules together with some physical-chemical factors. A new analytical method for identification and quantification of biologically active substances farnesol and tyrosol using ultra high performance liquid chromatography (UHPLC) in connection with tandem mass spectrometry was developed. The analytes were separated on Acquity BEH C18 analytical column using binary mobile phase consisting of acetonitrile and formic acid 0.075% (75:25) at flow-rate 0.20 ml/min. SRM (selected reaction monitoring) mode was applied in order to ensure sufficient selectivity and sensitivity using the first most intensive transition as a quantitative (121>77 and 205>121) and second one for the confirmation purposes (121>93 and 205>109). The method was validated in terms of linearity (>0.9994), precision (0.5-3.8% RSD), accuracy (78.9-106.0%), LOD (limit of detection) and LOQ (limit of quantitation). The method can serve as an analytical tool for the detection and determination of quorum-sensing molecules in biological samples.

Immunotoxicity activity of the major essential oil of Filipendula glaberrima against Aedes aegypti L.

The aerial parts of Filipendula glaberrima were extracted and the composition and immunotoxicity effects of major essential oils were studied. The analyses conducted by gas chromatography and mass spectroscopy (GC-MS) revealed the essential oils of F. glaberrima. The F. glaberrima essential oil (FGEO) yield was 0.046%, and GC/MS analysis revealed that its major constituents were ß-farnesol (2.96%), l-α-terpineol (2.43%), benzenemethanol (2.87%), (Z)-3-hexen-1-ol (5.23%), and 2,6-bis(1,1-dimethylethyl)-4-methylphenol (1.91%). The essential oil had a significant toxic effect against early fourth stage larvae of Aedes aegypti L with an LC(50) value of 28.43 ppm and an LC(90) value of 76.21 ppm. The results could be useful in search for newer, safer, and more effective natural immunotoxicity agents against A. aegypti.

[Chemical constituents from the seed of Chimonanthus praecox extracted by supercritical carbon dioxide].

OBJECTIVE: To analyse the volatile oil from seed of Chimonanthus praecox. METHODS: The volatile oil was extracted by Supercritical CO2 fluid and was analysed by GC-MS. RESULTS: Through comparison with mass spectra database, 43 components were isolated and identified. The main components are Calycanathine (51.88%), Calycanthidine (11.43%), n-Pentatriacontene (11.49%), (E, E)-2,4-Decadienal (9.67%), 1,1-diethoxyethoxy-3, 7-dimethyl-2, 6-Octadiene ( 3.40%), 7, 10-Octadecadienoic acid, methyl ester (3.38% ), 2, 13-Octadecadien-1-ol (2.90%), (1S-cis)-1,2,3,5,6,8a-hexahydro-4,7-dimethyl-1 (1-methylethyl)-Naphthalene (1. 90%), 3-Hexadecyne ( 1.82%), Hexadecanoic acid metyl ester( 1.78% ), Caryophyllene oxide (1.49%), n-Hexadecanoic acid (1.48%), Spathulenol (1.07%). CONCLUSION: Compounds (1S-cis)-1, 2,3,5,6,8a-hexahydro-4,7-dimethyl-1-(1-metyl-ethyl)-Naphthalene, Farnesol and Spathulenol are isolated from the Chimonanthus praecox for the first time.

Analytical method for determination of allylic isoprenols in rat tissues by liquid chromatography/tandem mass spectrometry following chemical derivatization with 3-nitrophtalic anhydride.

A liquid chromatography-tandem mass spectrometric (LC-MS/MS) method following chemical derivatization with 3-nitrophtalic anhydride was developed for the simultaneous determination of farnesol and geranylgeraniol in rat liver and testis. One analogue compound of the analytes, n-pentadecanol, was used as an internal standard (IS) for both analytes in this method. Rat tissues were disintegrated with 8% KOH ethanol solution, and then farnesol, geranylgeraniol and IS were extracted with a mixture of n-hexane-ethanol (98.5:1.5, v/v) in twice. Farnesol, geranylgeraniol and IS were then converted to 3-nitrophtalic derivatives of each analyte, and extracted with n-hexane. A turbo ion spray interface was used as the ionization source of LC-MS/MS and the analysis was performed in the multiple reaction monitoring (MRM) mode. The calibration curve at the spiked concentrations of 0.15-15 microg/g for both analytes showed good linearity. The method was precise; the relative standard deviations of the method for rat liver were not more than 13.4 and 5.4% for farnesol and geranylgeraniol, respectively, and those for rat testis were not more than 8.4 and 8.6% for farnesol and geranylgeraniol, respectively. The accuracies of the method for both rat liver and testis were good, with the deviations between the nominal concentration and calculated concentration of farnesol and geranylgeraniol typically being within 12.3 and 10.2%, respectively. This method provided reliable concentration levels for farnesol and geranylgeraniol in rat liver and testis.

Isolation and characterization of a monoamine oxidase B selective inhibitor from tobacco smoke.

It is well established that tobacco smokers have reduced levels of monoamine oxidase activities both in the brain and peripheral organs. Furthermore, extensive evidence suggests that smokers are less prone to develop Parkinson's disease. These facts, plus the observation that inhibition of monoamine oxidase B protects against the parkinsonian inducing effects of the nigrostriatal neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, have prompted studies to identify monoamine oxidase inhibitors in the tobacco plant and tobacco cigarette smoke. Our previous efforts on cured tobacco leaf extracts have led to the characterization of 2,3,6-trimethyl-1,4-naphthoquinone, a non-selective monoamine oxidase inhibitor, and farnesylacetone, a selective monoamine oxidase B inhibitor. We now have extended these studies to tobacco smoke constituents. Fractionation of the smoke extracts has confirmed and extended the qualitative results of an earlier report [J. Korean Soc. Tob. Sci.1997, 19, 136] demonstrating the inhibitory activity of the terpene trans,trans-farnesol on rat brain MAO-B. In the present study, K(i) values for the inhibition of human, baboon, monkey, dog, rat, and mouse liver MAO-B have been determined. Noteworthy is the absence of inhibitory effects on human placental MAO-A and beef liver MAO-B. A limited structure-activity relationship study of analogs of trans,trans-farnesol is reported. Although the health hazards associated with the use of tobacco products preclude any therapeutic opportunities linked to smoking, these results suggest the possibility of identifying novel structures of compounds that could lead to the development of neuroprotective agents.

Enhanced production of farnesol by Candida albicans treated with four azoles.

The dimorphic fungus Candida albicans excretes farnesol, which is produced enzymatically from the sterol biosynthetic intermediate farnesyl pyrophosphate. Inhibition of C. albicans by four azole antifungals, fluconazole, ketoconazole, miconazole, and clotrimazole, caused elevated farnesol production (10- to 45-fold). Furthermore, farnesol production occurs in both laboratory strains and clinical isolates (J. M. Hornby et al., Appl. Environ. Microbiol. 67:2982-2992, 2001) of C. albicans.

Chemical ecology of oribatid mites III. Chemical composition of oil gland exudates from two oribatid mites, Trhypochthoniellus sp. and Trhypochthonius japonicus (Acari: Trhypochthoniidae).

The composition of oil gland exudates from two oribatid mites, Trhypochthoniellus sp. and Trhypochthonius japonicus, was studied with reference to the related species Trhypochthoniellus crassus. Trhypochthoniellus sp. contained a mixture of seven compounds; (Z,Z)-6,9-heptadecadiene, geranial, 3-hydroxybenzene-1,2-dicarbaldehyde (gamma-acaridial), neryl formate, neral, (Z)-8-heptadecene and geranyl formate in decreasing order of abundance. The profile of the components from T. japonicus consisted of two types depending on the locality of sampling with unknown reason; one possessing a mixture of eight compounds [(Z,E)-farnesal, gamma-acaridial, (Z,Z)-6,9-heptadecadiene, (E,E)-farnesal, (Z)-8-heptadecene and geranial in decreasing order] together with two unknown compounds, and the other composed of the same set of compounds together with 2-hydroxy-6-methylbenzaldehyde as the most abundant component. Relative abundance among common components was consistent between the two types of T. japonicus. Profiles of components differed among three species including T. crasus. The phylogenetic relationship between Oribatida and Astigmata was discussed based on secretory compounds commonly distributed between these two suborders.