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Effect of dentin primers on the morphology, molecular composition and collagen conformation of acid-demineralized dentin in situ.

OBJECTIVES: The purpose of this study was to assess the effect of two commercial primers and distilled water on the morphology, molecular composition and collagen conformation of acid-demineralized dentin in situ. METHODS: Dentin specimens etched with Scotchbond Etchant were imaged by tapping mode AFM and analyzed by MIR-FTIR spectroscopy. They were then primed with Scotchbond Multi-Purpose Plus Primer, Scotchbond 1 Adhesive or distilled water and imaged and analyzed again. The chemical modifications induced on the uppermost 2 microns of primed dentin were studied after water and original primer subtraction. The conformational changes of type I collagen at this region were evaluated by deconvoluting the amide I band components. The absorbance ratio An(1655/1627) was used to semiquantitatively assess, on a relative basis, the extent of collagen denaturation. RESULTS: All the priming treatments swelled the collapsed dentin collagen left after etching. No evidence of primary bonding was found after priming treatments, while approximately 50% of the conditioned dentin collagen appeared denatured. Treatment with distilled water did not change the status of denatured collagen, however, application of the commercial primers refolded the alpha-helix to approximately 95% of the extent found on the native reference dentin. SIGNIFICANCE: The dynamic response of dentin collagen to demineralization and priming treatments reveals the critical role of some primers in rapidly restoring the conformational status of acid-denatured collagen. Implementation of reactive adhesive groups in alpha-helix recovery may provide an associative means of modifying the mechanical properties of the demineralized collagen based on the extent of their intermolecular bonding.

Acid Etching, Dental↗

Effects of flavor salience on aversion performance following relatively long-delay backward conditioning procedures.

Male albino rats (n = 144) received a 0.15 M injection of lithium chloride (at 2.0% body wt), followed 10, 30, or 75 min later by a 5.0% casein CS or a 10.0% sucrose CS, casein being the more salient CS. For each CS one-third of the rats received no fluid during the toxin-CS interval. The remaining two-thirds of the rats received 2-min access to distilled water or to a novel flavor 5 min after onset of the toxin-CS interval. For sucrose CS groups, the novel flavor was casein; for casein CS groups, the novel flavor was sucrose. Groups which received no fluid during the toxin-CS interval showed reliably greater aversion effects to the casein CS than to the sucrose CS. Results of Test Trial 1 showed that aversion to casein was relatively unchanged across toxin-CS intervals, while aversion to sucrose decreased reliably from the 10-min to the 30- and 75-min intervals. However, for each toxin-CS interval, aversion to the sucrose CS was reliably enhanced when casein access occurred in the interval, relative to that for distilled water or no fluid access. For the casein CS, access to sucrose or distilled water in the toxin-CS interval altered aversion effects, relative to the no fluid condition, depending on the interval.

Animals↗

Mutagenicity and contents of polycyclic aromatic hydrocarbons in used and recycled motor oils.

Thirteen samples of used motor oil and 33 recycled fractions, obtained in the laboratory by means of a recovery process similar to that currently used in Italy (vacuum distillation followed by thermal clay treatment) were examined. The Ames test (standard and modified version according to Blackburn) was used to determine the mutagenicity of the extracts and their contents of polyaromatic fraction (PAF) (IP346/80 method) and polycyclic aromatic hydrocarbons (PAH) (Grimmer's method). Used motor oils are mutagenic, both directly and indirectly. The highest values have been found in used oils from motor vehicles using leaded petrol (up to 118.8 revertants/mg). Samples from vehicles using unleaded petrol or diesel fuel are less mutagenic (up to 31.1 and 16.4 rev/mg, respectively). The enrichment in mutagens due to the use of oil in the three types of engine ranges from mean values of 6.2, 1.1 and 0.4 rev/mg per 1000 km, respectively. Recycled oils are almost completely devoid of direct mutagenic activity (33 samples: mean +/- SD = 1.6 +/- 1.5 rev/mg). Most recycled distillates show considerable mutagenic activity in the presence of microsomial enzymes (up to 82.5 rev/mg), although this is reduced with respect to the original oils (recycled, mean +/- SD = 13.8 +/- 15.5 rev/mg; original oils, mean +/- SD = 30.7 +/- 35.2, Mann-Whitney U-test, z = 1.793, p < 0.05). Both PAF and PAH contents are high in used oils from the two types of petrol engine but not in those from diesel engines. Recycling reduces PAF contents only in used oils from petrol engines, from a mean value of 13.91 +/- 7.32 to 4.23 +/- 2.90% (comparison with original used oils, Mann-Whitney U-test, U = 8, p < 0.01). The light distilled fractions have greater concentrations of indirect mutagens, PAF and PAH than the others. The increase in PAH in light recycled products with respect to the original used oils is significant (Wilcoxon's t-test, z = 2.306, p < 0.05). Benzo[a]pyrene (BaP) is found in appreciable quantities (> 10 ppm) in all used oils from petrol engines and in most of their recycled products. Recycling generally recovers 50% of mutagens and PAF and about 80% of PAH. Considered together, recycled products have in any case contents of mutagens and PAF which are significantly lower than those in the parent oils, but not of PAH (Wilcoxon's t-test; mutagens, z = 2.935, p < 0.01; PAF, z = 3.145, p < 0.01; PAH, z = 1.397, not significant). Lastly, many recycled oils have PAH concentrations which are equal to or higher than those of the original used oils. The health risks linked to professional exposure to these types of oils and the inadequate recycling process currently used (redistillation and thermal clay treatment) in reducing mutagenic and cancerogenic substances from used motor oils are stressed.

Fuel Oils↗

Suppressive effect of irsogladine maleate on N-methyl-N-nitro-N-nitrosoguanidine (MNNG)-initiated and glyoxal-promoted gastric carcinogenesis in rats.

The modifying effect of irsogladine maleate (IRG) on N-methyl-N-nitro-N-nitrosoguanidine (MNNG)-initiated and glyoxal-promoted gastric carcinogenesis was examined in male Wistar rats. Six-week-old rats were divided into ten groups. Groups 1 through 6 were given MNNG (100 mg/l in drinking water) for 25 weeks from the start of the experiment, whereas groups 7 through 10 received distilled water in the initiation phase as the vehicle treatment. Groups 1 and 8 were kept on the basal diet and distilled water throughout the experiment (55 weeks). Groups 2-8 were given 0.5% glyoxal in the drinking water for 30 weeks from 26th week of the experiment. Group 3 was fed the diet mixed with 100 ppm IRG for 25 weeks from the start of experiment. Groups 4 and 8 were fed the diet mixed with 100 ppm IRG for 30 weeks from 26th week of experiment. Groups 5 and 9 or 6 were given 100 or 25 ppm IRG containing diet, respectively throughout the experiment. Group 10 was given the basal diet and distilled water as the vehicle treated control. Tumors of upper digestive tracts (stomach and duodenum) were developed in groups: 1 (12/17 rats, 71%), 2 (11/12 rats, 92%), 3 (9/16 rats, 56%), 4 (5/12 rats, 42%), 5 (6/15 rats, 40%) and 6 (7/12 rats, 58%). High dose of IRG in initiation and/or promotion phase significantly reduced the incidence of tumors of the upper digestive tracts. The average numbers of the digestive tracts neoplasms in groups 3,5 and 6 given glyoxal and IRG were less than those in group 2 which received only glyoxal. These results suggest that IRG could be a preventive agent against the occurrence of neoplasms of the upper digestive tract.

Animals↗

Water uptake of soft lining materials from osmotic solutions.

OBJECTIVES: The water uptake characteristics of soft lining materials are of obvious importance in that they are expected to function in the oral environment. Results for Novus (Hygenic Corp., Akron, OH, USA) show a very high uptake from distilled water. Despite this high uptake, Novus appears to function satisfactorily in the mouth. High water uptake of soft lining materials has been attributed to the presence of water soluble impurities that, on immersion, form solution droplets; the driving force for the uptake being the osmotic gradient between the droplets and the external solution. Uptake should therefore be less from ionic solutions. The object of this study was to test the applicability of this theory to Novus and two experimental soft lining materials. METHODS: Water uptake of two experimental materials and Novus has been determined from distilled water and two saline solutions (0.45 and 0.9 M). After 196 days specimens were desorbed to constant weight and then subjected to a second sorption cycle. RESULTS: Novus had the highest uptake from distilled water at approximately 18%, the experimental materials having an uptake approximately 7%. Desorptions were all rapid, minimum weight being reached within 1-2 days. Uptakes of the second sorptions from water were all higher. Uptake from saline solutions was approximately 12% for all materials, uptake from 0.9 M saline being the lowest. Second sorption results from solution were similar to the first. CONCLUSION: The results obtained support the theory that the high water uptake of elastomeric materials is osmotically driven.

Adsorption↗

Long-term surface micro-hardness of resin-modified glass ionomers.

OBJECTIVES: The presence of a resin component in resin-modified glass-ionomer cements (RMGICs) results in considerable water sorption which may affect the surface hardness of the materials. This study investigated the effect of long-term storage in aqueous solutions on the surface hardness of RMGICs. METHODS: The surface micro-hardness of four RMGICs stored in either distilled water or artificial saliva was measured using the Wallace microindentation tester at regular time intervals up to 360 days. RESULTS: There was a substantial increase in hardness during the first day of storage when RMGICs were kept in distilled water. All RMGICs except one liner/base material reached their maximum hardness within 7 days and maintained their hardness for up to 1 year. RMGICs stored in artificial saliva were relatively softer and showed a decrease in surface hardness with time. CONCLUSIONS: The post-hardening reaction overcame the plasticising effect of water when RMGICs were stored in distilled water. However, progressive and high water uptake of specimens stored in artificial saliva resulted in a decrease in their surface hardness.

Analysis of Variance↗

In vitro fluoride release from restorative materials in water versus artificial saliva medium (SAGF).

OBJECTIVES: An extensive number of investigations have focused in recent years on the process of fluoride release by glass-ionomer cements. In order to compare the fluoride release of two recent resin-modified glass-ionomer cements (RMGICs) and one compomer, we investigated the initial and subsequent fluoride release in distilled water and in a mineral medium with composition similar to saliva (SAGF), as well as the renewal effect of the media on the release. METHOD: A preliminary thermodynamic study defined conditions under which the measurements by fluoride-selective electrode potentiometry were not affected by the presence of Ca2+ ions in SAGF. Disk-shaped samples (5 mm x 3 mm) for each material were placed in 5 ml of distilled water (n = 12) or 5 ml of SAGF (n = 12) and fluoride released was measured after 1, 2 and 7 days. For the investigation of medium renewal effect, RMGIC sample were dipped into distilled water for 5 days. Subsequently 12 samples of each material were placed for 7 days into water or SAGF, with or without daily renewal of the medium before fluoride-release measurements. The elements present on the surface of the RMGICs were analyzed by scanning electron microscopy. RESULTS: RMGICs released significantly more fluoride in water than in artificial saliva. The presence of CaF2 at the interface accounts for this phenomenon. The thickness of the CaF2 layer depends on the speed of its initial formation and renewal of the testing medium. A pattern of CaF2 formation was proposed.

Calcium Fluoride↗

Fatty acid compositions in local sea cucumber, Stichopus chloronotus, for wound healing.

Fatty acid profile from crude extracts of local sea cucumber Stichopus chloronotus was determined using gas chromatography (GC) technique. The extracts were prepared separately in methanol, ethanol, phosphate buffer saline (PBS), and distilled water as part of our study to look at the affinity of these solvents in extracting the lipid from sea cucumber. The PBS and distilled water extractions indicate water-soluble components, while the organic fractions are extracted in methanol and ethanol as organic solvents. Furthermore, water extraction is the conventional method practiced in Malaysia. In our analysis the C14:0 (myristic), C16:0 (palmitic), C18:0 (stearic), C18:2 (linoleic), C20:0 (arachidic), and C20:5 (eicosapentaenoic, EPA) were significantly different (p < 0.01) in the four solvent extractions. However, the PBS extraction contained a much higher percentage of EPA (25.69%) compared to 18.89% in ethanol, 7.84% in distilled water, and only 5.83% in methanol, and variances were significantly different (p < 0.01 ). On the other hand, C22:6 (docosahexaenoic acid or DHA) is much higher in water extraction (57.55%), in comparison to the others where only 3.63% in PBS and 1.20% in methanol, and this difference is significant at p < 0.01. No DHA was detected in ethanol extractions. Subsequently, C18:1 (oleic acid) was only detected in PBS (21.98%) and water extraction (7.50%). It is interesting that palmitic acid, C16:() was higher in methanol (20.82%) and ethanol (2.18%), while 12.55% was detected in PBS and only 2.20% in water extraction: and again this was significantly different at p < 0.01. Although our results have shown that all four solvents were different in terms of their ability to extract fatty acids, the major component for tissue repair was well preserved. Probably this is one of the important precocious steps when working with a delicate sea cucumber, in both experimental and/or at the preparative stages. Freshness of the sea cucumber samples is important when undertaking this type of experiment. Finally, we believe that the local sea cucumber S. chloronotus contains all the fatty acids required to play a potential active role in tissue repair.

Animals↗

Selective activity of Streblus asper on Mutans streptococci.

The purpose of this in vivo one group time series design and single blind study was to determine the antimicrobial effectiveness of a mouthrinse containing Streblus asper leaf extract on Streptococcus mutans and total salivary bacteria following single 60 s rinse. Changing in salivary pH and buffer capacity during the experimental period were also measured. Thirty human subjects participated in this study. At each experimental session, a pretest saliva sample was taken. After giving the pretest samples, the subjects rinsed with S. asper leaf extract or distilled water control for 60 s, then the post-test saliva samples were collected at 0, 0.5, 1, 3, 5 and 6 h. The samples were used to determine the numbers of S. mutans and total salivary bacteria. The results indicated that S. asper leaf extract significantly reduced S. mutans counts compared with distilled water. However, the mean difference from baseline of total salivary bacterial counts was not significantly different between rinsing with S. asper leaf extract and distilled water. Moreover, S. asper leaf extract showed no effects in modifying the salivary pH and buffer capacity. It is concluded that of mouthrinse containing S. asper leaf extract can reduce S. mutans without changing an oral ecology.

Adolescent↗

Some optimisation studies relevant to the production of high-purity 124I and 120gI at a small-sized cyclotron.

Optimisation experiments on the production of the positron emitting radionuclides 124I(T(1/2) = 4.18d) and (120g)I (T(1/2) = 1.35 h) were carried out. The TeO(2)-target technology and dry distillation method of radioiodine separation were used. The removal of radioiodine was studied as a function of time and the loss of TeO(2) from the target as a function of oven temperature and time of distillation. A distillation time of 15 min at 750 degrees C was found to be ideal. Using a very pure source and comparing the intensities of the annihilation and X-ray radiation, a value of 22.0 +/- 0.5% for the beta(+) branching in 124I was obtained. Production of 124I was done using 200 mg/cm(2) targets of 99.8% enriched 124TeO(2) on Pt-backing, 16 MeV proton beam intensities of 10 microA, and irradiation times of about 8 h. The average yield of 124I at EOB was 470 MBq(12.7 mCi). At the time of application (about 70 h after EOB) the radionuclidic impurity 123I (T(1/2) = 13.2 h) was <1%. The levels of other impurities were negligible (126I < 0.0001%;125I = 0.01%). Special care was taken to determine the 125I impurity. For the production of (120g)I only a thin 30 mg target (on 0.5 cm(2) area) of 99.9% enriched 120TeO(2) was available. Irradiations were done with 16 MeV protons for 80 min at beam currents of 7 microA. The 120gI yield achieved at EOB was 700 MBq(19 mCi), and the only impurity detected was the isomeric state 120 mI(T(1/2) = 53 min) at a level of 4.0%. The radiochemical purity of both 124I and 120gI was checked via HPLC and TLC. The radioiodine collected in 0.02 M NaOH solution existed >98% as iodide. The amount of inactive Te found in radioiodine was <1 microg. High purity 124I and 120gI can thus be advantageously produced on a medium scale using the low-energy (p,n) reaction at a small-sized cyclotron.

Cyclotrons↗

Dentin penetration of 2 calcium hydroxide combinations.

OBJECTIVE: Calcium hydroxide is a common intracanal medicament. The powder can be mixed with different vehicles for placement in canals. This study examines pH changes in dentin with 2 different calcium hydroxide preparations. STUDY DESIGN: The combinations used were calcium hydroxide + distilled water and calcium hydroxide + glycerin (1/7 distilled water). One hundred forty extracted teeth with single canals were used. pH changes in dentin after the intracanal placement of these combinations were evaluated at different dentin thicknesses by measuring the pH of the external environment. RESULTS: For all dentin thicknesses and at all time periods, the calcium hydroxide + glycerin combination showed significantly higher pH values than the calcium hydroxide + distilled water combination. CONCLUSIONS: Considering pH elevations, we propose the use of a calcium hydroxide + glycerin combination.

Calcium Hydroxide↗

Distribution of cholesterol in milk fat fractions.

Milk fat was fractionated into liquid (m.p. congruent to 12 degrees C), intermediate (m.p. congruent to 21 degrees C) and solid (m.p. congruent to 39 degrees C) fractions by three different processes--melt crystallization, short-path distillation and supercritical CO2 extraction--and the cholesterol content of these fractions determined. Cholesterol was enriched in the liquid fractions from all three processes, in particular about 80% of the cholesterol being found in the liquid fraction obtained by short-path distillation. The basis of migration of cholesterol into various milk fat fractions was explained by its affinity to various triglycerides (melt crystallization) and by vapour pressure and molecular weight (short-path distillation). It was more complex in the supercritical CO2 extraction process; the interplay of cholesterol affinity toward CO2 and its molar volume, and its vapour pressure enhancement under applied pressure play a role.

Animals↗

Astatine-211 labeling of an antimelanoma antibody and its Fab fragment using N-succinimidyl p-astatobenzoate: comparisons in vivo with the p-[125I]iodobenzoyl conjugate.

Astatine-211 labeling of an antimelanoma antibody, NR-ML-05, and its Fab fragment with N-succinimidyl p-[211At]astatobenzoate (2a) has been described. Preparation of the astatinated intermediate 2a was accomplished by distilling astatine-211 from an irradiated bismuth target directly into a reaction mixture containing an organometallic compound, N-succinimidyl p-(tri-n-butylstannyl)benzoate (1), and an oxidant, N-chlorosuccinimide, in 5% HOAc/MeOH. Trapping of distilled astatine as 2a was found to be efficient, resulting in 70-90% yields based on the amount of astatine-211 in the reaction mixture. The dry distillation technique employed gave recoveries of astatine-211 which ranged from 20% to 75%. Conjugation of 2a to NR-ML-05 and its Fab fragment was accomplished in 40-60% yields. The [211At]astatobenzoyl-conjugated antibodies were found to be stable in vitro when challenged by strong denaturants and nucleophilic reagents. Coinjected dual-labeled studies of the 2a astatinated antibodies and the same antibodies labeled with N-succinimidyl p-[125I]iodobenzoate (2b) in athymic mice bearing the human tumor xenograft A375 Met/Mix demonstrated that both radiolabeled antibodies had equivalent tumor localization. Data from the dual-labeled biodistribution of the intact antibody suggests that the astatine is stably attached. Data from the dual-labeled Fab fragment suggests that a portion of the astatine label is released as astatide, either from the astatinated Fab or from a catabolite.

Animals↗

Toxic effects of unresolved complex mixtures of aromatic hydrocarbons accumulated by mussels, Mytilus edulis, from contaminated field sites.

Exposure of marine mussels (Mytilus edulis) to an unresolved complex mixture (UCM) of aromatic hydrocarbons isolated from a crude oil has been shown to reduce their feeding rate by 40%. The present study was undertaken to determine whether UCMs bioaccumulated by mussels in the field are also toxic. The feeding rate of mussels derived from polluted sites increased when they were placed in clean water, pointing to a loss of toxic agents from the tissues. At the end of the depuration period, water in which mussels from an oil-polluted site had been held contained a UCM. Steam-distillation extracts of the tissues of mussels taken from several polluted sites were shown to be highly toxic to the feeding activity of juvenile mussels. The tissues of mussels from these sites contained UCMs. Nontoxic steam distillates from clean mussels did not. Steam-distillation extracts of mussels from an oil-polluted site were fractionated by normal-phase high-performance liquid chromatography. A fraction, largely comprising a "monoaromatic" UCM, reduced the feeding rate of juvenile mussels by 70%. Two later-eluting fractions containing aromatic UCMs also produced smaller depressions in feeding rate. These results support our contention that some aromatic UCM hydrocarbons constitute a forgotten pollutant burden in the marine environment.

Animals↗

Evaluation and testing of analytical methods for cyanide species in municipal and industrial contaminated waters.

Total cyanide analysis by distillation is used most commonly to assess cyanide content of water samples. This manual method is robust but slow and provides no information about cyanide speciation, a significant limitation in that cyanide species have substantially different toxicity characteristics. Seven alternative methods for the analysis of cyanide species or groups of species were evaluated in reagent water and five different contaminated water matrices, including five species-specific methods--weak acid dissociable (WAD) cyanide, free cyanide by microdiffusion, available cyanide, automated WAD cyanide by thin film distillation, metal cyanides by ion chromatography--and two automated techniques for total cyanide--total cyanide bythin film distillation and total cyanide by low-power UV digestion. The species-specific cyanide analytical techniques achieved low, ppb-level detection limits and exhibited satisfactory accuracy and precision for most contaminated waters. Analysis of low concentrations of cyanide species in raw wastewater was problematical for the available cyanide and ion chromatography methods, which experienced significant interference problems and/or low recoveries. There was recovery of significant diffusible cyanide in microdiffusion tests with nickel-cyanide-spiked samples, reflecting dissociation of this weak metal-cyanide complex during the test and demonstrating that the test can recover some fraction of WAD cyanide in addition to free cyanide. The automated total cyanide methods, which involve UV digestion, achieved low detection limits for most waters but exhibited low recoveries for some waters.

Chromatography, Ion Exchange↗

Environmental assessment of used oil management methods.

The 1 billion gal of used oil generated in the U.S. each year are managed in three primary ways: rerefined into base oil for reuse, distilled into marine diesel oil fuel, and marketed as untreated fuel oil. Management of used oil has local, regional and global impacts. Because of the globally distributed nature of fuel markets, used oil as fuel has localized and regional impacts in many areas. In this paper, the human health and environmental tradeoffs of the management options are quantified and characterized. The goal of this study was to assess and compare the environmental impacts and benefits of each management method in a product end-of-life scenario using a life-cycle assessment (LCA) approach. A life-cycle inventory showed that 800 mg of zinc and 30 mg of lead air emissions may result from the combustion of 1 L of used oil as fuel (50-100 times that of crude-derived fuel oils). As an example, up to 136 Mg of zinc and 5 Mg of lead air emissions may be generated from combustion of over 50 M gal of California-generated used oil each year. While occurring elsewhere, these levels are significant (of the same magnitude as reported total stationary source emissions in California). An impact assessment showed that heavy metals-related toxicity dominates the comparison of management methods. Zinc and lead emissions were the primary contributors to the terrestrial and human toxicity impact potentials that were calculated to be 150 and 5 times higher, respectively, for used oil combusted as fuel than for rerefining or distillation. Low profits and weak markets increasingly drive the used oil management method selection toward the untreated fuel oil market. Instead, both the rerefining and distillation methods and associated product markets should be strongly supported because they are environmentally preferable to the combustion of unprocessed used oil as fuel.

Air Pollutants↗

Determination of bisphenol A in food-simulating liquids using LCED with a chemically modified electrode.

Liquid chromatography with electrochemical detector (LC-ED), using a chemically modified electrode coated with a metalloporphyrin film, is reported for determination of bisphenol A (BPA) migration from polycarbonate baby bottles. The extraction process of the samples was performed according to regulations of the Southern Common Market (MERCOSUR), where certain food-simulating liquids [(A) distilled water, (B) acetic acid 3% V/V in distilled water, and (C) ethanol 15% V/V in distilled water] are defined along with controlled time and temperature conditions. The baseline obtained using the naked electrode showed a considerable drift which increased the detection limit. This effect was suppressed with the chemically modified electrode. A linear range up to 450 ppb along with a detection limit of 20 ppb for the amperometric detection technique was observed. The procedure described herein allowed lowering the detection limit of the method to 0.2 ppb. The value found for BPA in the food-simulating liquid is 1.2 ppb, which is below the tolerance limit for specific migration (4.8 ppm).

Benzhydryl Compounds↗

Supercritical carbon dioxide extraction of essential oils from Perovskia atriplicifolia Benth.

The supercritical fluid extraction (SFE) of the aerial parts of Perovskia atriplicifolia Benth. was studied. The effect of different parameters such as pressure, temperature, modifier identity, and modifier volume on the SFE of the plant was investigated. The extracts were analyzed by GC and GC-MS and compared with the essential oil obtained from P. atriplicifolia Benth by steam distillation. The supercritical extracts and the steam-distilled products had very different compositions. The main constituents of the oil obtained by steam distillation were 1,8-cineole, limonene, camphor, beta-caryophyllene, alpha-pinene, camphene, and alpha-humulene. On the other hand, the major components of SFE extracts were 1,8-cineole, limonene, camphor, beta-caryophyllene, gamma-cadinene, alpha-pinene, and alpha-terpinyl acetate. The results showed that increasing the temperature from 35 to 65 degrees C (at a constant pressure of 100 atm) drastically reduced the number of extracted components. Also, the number of extracted constituents and the percent of main analytes increased when lower pressures were used. Using different modifiers (e.g., methanol, ethanol, dichloromethane, and hexane) for the extraction of the plant at low pressure (100 atm) and temperature (35 degrees C) showed that hexane was more selective than the other modifiers.

Bicyclic Monoterpenes↗