Extraction of Natural Fragrance Ingredients: History Overview and Future Trends.

For centuries, perfumes consisted in a combination of natural ingredients, mainly of plant origin. From the 19th century on, the advent of organic synthesis enabled the deployment of multiple synthetic olfactory notes, enriching significantly the perfumers' portfolio. Chemistry is ever since the foundation of modern perfumery. However, sustainable-minded consumers, massively rejecting synthetics for safety and ecological issues, engaged a global return to nature in numerous sectors, and the fragrance industry is not outdone. Sustainable extraction of natural products, making use of innovative technologies, process intensification and agro-based solvents, constitutes the answer to develop eco-conceived fragrant ingredients covering every olfactory family without endangering biodiversity any further. The objective of this review is to draw a clear picture of where those technological advances are today and to assess the ones that may be effectively transposed at the industrial scale tomorrow.

Electrochemical preparation of poly(3-methylthiophene-carbazole)/graphene oxide composite coating for the highly effective solid-phase microextraction of some fragrance.

A poly(3-methylthiophene-carbazole)/graphene oxide composite coating was prepared on a stainless steel wire through electrochemical method. It showed high extraction efficiency, thermal stability and durability than those of corresponding single-component coatings due to the synergistic effect of different components. The resulting fiber was used for the headspace solid phase microextraction of octanal, nonanal, decanal, undecanol and dodecanol, followed by gas chromatography detection. The results showed that their GC peak areas were linear to their concentrations in the ranges of about 0.05-100µgL-1, the detection limits were 22.6-40.5ngL-1 (S/N=3). The run-to-run RSD was smaller than 4.5% (n=4), and the fiber-to-fiber RSD was 4.0-9.3% (n=4). The method was successfully applied to the determination of the five analytes in candy, orange fruit and flower.

Degradation of synthetic fragrances by laccase-mediated system.

Laccase mediator systems are important biodegradation agents as the rate of reaction could be enhanced in the presence of redox mediators. In the present study the commercial enzyme laccase from Trametes versicolor and the redox mediator 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) were used for the biotransformation of the synthetic fragrances 1-(1,2,3,4,5,6,7,8-octahydro-2,3,8,8,-tetramethyl-2-naphthyl)ethan-1-one (Iso-E-Super, OTNE), 1,3,4,6,7,8,-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta-[g]-2-benzopyran (Galaxolide, HHCB), 7-acetyl-1,1,3,4,4,6-hexamethyl-1,2,3,4-tetrahydronaphtalene (Tonalide, AHTN) and the transformation product of HHCB, 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta-[g]-2-benzopyran-1-one (Galaxolidone, HHCB-lactone) in water. A particular focus was to assess the effects of the enzyme laccase from Trametes versicolor in the enantioselective degradation of the target compounds, for this reason gas chromatography with an enantioselective column was used as separation technique followed by mass spectrometry detection. In addition, as enantioselective degradation of musk fragrances was observed in wastewater, sewage sludge and fish samples, enantiomeric fractions of selected compounds were studied during composting. In a period of 144h, the target fragrances could be effectively removed by the enzyme laccase with removal percentages greater than 70%, except AHTN with a removal percentage of 42%. However, the degradation process prompted by the enzyme laccase was shown to be non-enantioselective as no significant differences were observer between the enantiomeric fractions calculated at the beginning and at the end of the degradation process. Meanwhile, the composting process was shown to be enantioselective.

Analysis of flavors and fragrances by HPLC with Fe3O4@GO magnetic nanocomposite as the adsorbent.

In this article, a method for the analysis of flavors and fragrances including ethyl vanillin, trans-cinnamic acid, methyl cinnamate, ethyl cinnamate, and benzyl cinnamate in foods by HPLC with Fe3O4@GO nanocomposite as the adsorbent of magnetic solid-phase extraction was developed. The magnetic Fe3O4@GO nanocomposite was prepared and characterized by transmission electron microscope, infrared spectroscopy, X-ray diffraction and vibrating sample magnetometer. The as-prepared Fe3O4@GO nanocomposite was used as adsorbent for extraction and preconcentration of flavors and fragrances in foods. The extraction and desorption conditions including amount of materials, extraction time and desorption solvents were investigated. The analytes were separated within 10min and detected with LODs ranged from 0.02 to 0.04µg/mL. The recovery obtained from the analysis of spiked juice sample was between 71.5% and 112.4% with RSDs lower than 4.14%. The developed method was successfully applied to the analysis of real samples including orange juice, chocolate and fruit sugar.

Opinion of the Scientific Committee on Consumer Safety (SCCS) - Opinion on the fragrance ingredients Tagetes minuta and Tagetes patula extracts and essential oils (phototoxicity only) in cosmetic products.

CONCLUSION OF THE OPINION: The SCCS considers a maximum level of 0.01% Tagetes minuta and Tagetes patula extracts and essential oils in leave-on products (except sunscreen cosmetic products) as safe, provided that the alpha terthienyl (terthiophene) content of the Tagetes extracts and oils does not exceed 0.35%. The Tagetes extracts and oils should not be used as ingredients of sunscreen products.

Screening of phthalate esters in 47 branded perfumes.

In the last few years, the use of phthalates in perfumes has gained attention because these chemicals are sometimes added intentionally as a solvent and a fixative. Five phthalate esters, dimethyl phthalate (DMP), diethyl phthalate (DEP), dibutyl phthalate (DBP), benzyl butyl phthalate (BBP), and diethyl hexyl phthalate (DEHP), were measured in 47 branded perfumes using headspace solid phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC-MS). The results revealed considerable amounts of phthalate in all 47 brands with detection frequencies > limit of quantitation in the following order: DEP (47/47) > DMP (47/47) > BBP (47/47) > DEHP (46/47) > DBP (23/45). Of the 47 brands, 68.1, 72.3, 85.1, 36.2, and 6.7 % had DEP, DMP, BBP, DEHP, and DBP levels, respectively, above their reported threshold limits. Of these phthalates, DEP was found to have the highest mean value (1621.625 ppm) and a maximum of 23,649.247 ppm. The use of DEP in the perfume industry is not restricted because it does not pose any known health risks for humans. DMP had the second highest level detected in the perfumes, with a mean value of 30.202 ppm and a maximum of 405.235 ppm. Although DMP may have some uses in cosmetics, it is not as commonly used as DEP, and again, there are no restrictions on its use. The levels of BBP were also high, with a mean value of 8.446 ppm and a maximum of 186.770 ppm. Although the EU banned the use of BBP in cosmetics, 27 of the tested perfumes had BBP levels above the threshold limit of 0.1 ppm. The mean value of DEHP found in this study was 5.962 ppm, and a maximum was 147.536 ppm. In spite of its prohibition by the EU, 7/28 perfumes manufactured in European countries had DEHP levels above the threshold limit of 1 ppm. The DBP levels were generally low, with a mean value of 0.0305 ppm and a maximum value of 0.594 ppm. The EU banned the use of DBP in cosmetics; however, we found three brands that were above the threshold limit of 0.1 ppm, and all were manufactured in European countries. The results of this study are alarming and definitely need to be brought to the attention of the public and health regulators. Although some phthalate compounds are still used in cosmetics, many scientists and environmental activists have argued that phthalates are endocrine-disrupting chemicals that have not been yet proven to be safe for any use, including cosmetics. Phthalates may also have different degrees of estrogenic modes of action. Furthermore, we should not dismiss the widespread use of phthalates in everyday products and exposure to these chemicals from sources such as food, medications, and other personal care products.

Genetic variability in G2 and F2 region between biological clones of human respiratory syncytial virus with or without host immune selection pressure

Human respiratory syncytial virus (HRSV) is an important respiratory pathogens among children between zero-five years old. Host immunity and viral genetic variability are important factors that can make vaccine production difficult. In this work, differences between biological clones of HRSV were detected in clinical samples in the absence and presence of serum collected from children in the convalescent phase of the illness and from their biological mothers. Viral clones were selected by plaque assay in the absence and presence of serum and nucleotide sequences of the G2 and F2 genes of HRSV biological clones were compared. One non-synonymous mutation was found in the F gene (Ile5Asn) in one clone of an HRSV-B sample and one non-synonymous mutation was found in the G gene (Ser291Pro) in four clones of the same HRSV-B sample. Only one of these clones was obtained after treatment with the child's serum. In addition, some synonymous mutations were determined in two clones of the HRSV-A samples. In conclusion, it is possible that minor sequences could be selected by host antibodies contributing to the HRSV evolutionary process, hampering the development of an effective vaccine, since we verify the same codon alteration in absence and presence of human sera in individual clones of BR-85 sample.

Removal of PPCPs from the sludge supernatant in a one stage nitritation/anammox process.

Pharmaceutical and personal care products (PPCPs) are extensively used and can therefore find their way into surface, groundwater and municipal and industrial effluents. In this work, the occurrence, fate and removal mechanisms of 19 selected PPCPs was investigated in an 'ELiminación Autótrofa de Nitrógeno' (ELAN) reactor of 200 L. In this configuration, ammonium oxidation to nitrite and the anoxic ammonium oxidation (anammox)processes occur simultaneously in a single-stage reactor under oxygen limited conditions. The ELAN process achieved high removal (>80%) of the studied hormones, naproxen, ibuprofen, bisphenol A and celestolide, while it was not effective in the removal of carbamazepine (<7%), diazepam (<7%) and fluoxetine (<30%). Biodegradation was the dominant removal mechanism, while sorption was only observed for musk fragrances, fluoxetine and triclosan. The sorption was strongly dependent on the granule size, with smaller granules facilitating the sorption of the target compounds. Increased hydraulic retention time enhanced the intramolecular diffusion of the PPCPs into the granules, and thus increased the solid phase concentration. The increase of nitritation rate favored the removal of ibuprofen, bisphenol A and triclosan, while the removal of erythromycin was strongly correlated to the anammox reaction rate.

The search for new odorants: synthesis of animalic fragrant and musky/ambery compounds.

An overview of the recent research which allowed us to discover novel animalic odorants is presented. The new derivatives were prepared from readily available starting materials via easy reaction steps in good yields. They possess very different structures, such as bicyclic pentanols, glycolates, or tricyclic ketones, and all show interesting notes in the animalic fragrant family: from costus, leathery to ambery and musky scents, making them all attractive for different purposes.

Changes in growth, essential oil yield and composition of geranium (Pelargonium graveolens L.) as affected by growing media.

BACKGROUND: Using proper growing medium is known to be an effective way to improve crop growth and yield. However, the effects of growing media on geranium essential oil have scarcely ever been examined in detail. In this research, the effects of different growing media (soil, sand, pumice, perlite and perlite + cocopeat) on growth, oil yield and composition of geranium were studied. RESULTS: Growth was significantly improved in soilless-grown plants compared with soil-grown plants. Oil yield of soilless-grown plants (except for pumice) was about threefold higher than that of soil-grown plants. The increase in oil yield was correlated with higher leaf dry weight (r² = 0.96), as oil content was not affected. The citronellol/geranium ratio of oil was clearly affected by growing media, ranging from 5:1 in soil culture to 3:1 in soilless culture. The latter is acceptable for perfumery. CONCLUSION: Compared with soil, soilless media could produce higher yields of high-quality geranium oil that fits market requirements. Growth, oil yield and composition of plants grown in sand (a cheap and abundant growing medium) were not significantly different from those of plants grown in perlite and perlite + cocopeat.

Analysis of plasticizers and synthetic musks in cosmetic and personal care products by matrix solid-phase dispersion gas chromatography-mass spectrometry.

Matrix solid-phase dispersion (MSPD) and gas chromatography-mass spectrometry were used for the rapid determination of 18 plasticizers (phthalates and adipates), 7 polycyclic musks and 5 nitromusks, which makes a total of 30 targets, in both rinse-off and leave-on cosmetic formulations. The MSPD method was miniaturized and customized to avoid or minimize risks of phthalate contamination and to reduce residues and costs. The amount of sample and extraction solvent employed were 0.1g and 1mL, respectively. The procedure was optimized by means of an experimental design and under the optima conditions it showed satisfactory linearity, repeatability and intermediate precision. LOQs were, in general, in the low ngg(-1), and recoveries were quantitative for all the 18 plasticizers and the 12 fragrances. Twenty-six cosmetic products such as creams, emulsions, lotions, gels for the skin, bath and shower preparations, deodorants, hair-setting, hair-cleansing and hair-conditioning products, shaving products, and sunbathing products, were analyzed. Twenty-five out of thirty targets were detected in the samples. The most frequently found compounds were galaxolide and tonalide reaching concentrations above 0.1% (1000µgg(-1)), and diethyl phthalate (between 0.7 and 357µgg(-1)). The presence of banned substances (Regulation (EC) No. 1223/2009) such as dibutyl phthalate, diisobutyl phthalate, dimethoxyethyl phthalate, benzylbutyl phthalate, diethylhexyl phthalate, diisopentyl phthalate and dipentyl phthalate, musk ambrette and musk tibetene was confirmed in sixteen of the twenty-six personal care products (62%).

Determination of musk fragrances in sewage sludge by pressurized liquid extraction coupled to automated ionic liquid-based headspace single-drop microextraction followed by GC-MS/MS.

A method for the quantitative determination of ten musk fragrances extensively used in personal care products from sewage sludge was developed by using a pressurized liquid extraction (PLE) followed by an automated ionic liquid-based headspace single-drop microextraction and gas chromatography-tandem mass spectrometry. The influence of main factors on the efficiency of PLE was studied. For all musks, the highest recovery values were achieved using 1 g of pretreated sewage sludge, H(2) O/methanol (1:1) as an extraction solvent, a temperature of 80°C, a pressure of 1500 psi, an extraction time of 5 min, 2 cycles, a 100% flush volume, a purge time of 120 s, and 1 g Florisil as in-cell clean-up extraction sorbent. The use and optimization of an in-cell clean-up sorbent was necessary to remove fatty interferents of the PLE extract that make the subsequent ionic liquid-based headspace single-drop microextraction difficult. Validation parameters, namely LODs and LOQs, ranged from 0.5-1.5 to 2.5-5 ng/g, respectively. Good levels of intra- and interday repeatabilities were obtained analyzing sewage sludge samples spiked at 10 ng/g (n = 3, RSDs < 10%). The method applicability was tested with sewage sludge from different wastewater treatment plants. The analysis revealed the presence of all the polycyclic musks studied at concentrations higher than the LOQs, ranging from 6 to 530 ng/g. However, the nitro musk concentrations were below the LOQs or, in the case of musk xylene, was not detected.

[Determination of 21 fragrance allergens in toys by gas chromatography-ion trap mass spectrometry].

A method of gas chromatography-ion trap mass spectrometry (GC-IT-MS) was developed for the determination of 21 fragrance allergens in sticker toys, plush toys and plastic toys. The experimental conditions, such as sample pretreatment conditions, and the analytical conditions of GC-IT-MS, were optimized. The sticker toy samples and plush toy samples were extracted with acetone by ultrasonic wave, and the extracts were separated on an Agilent HP-1 MS column (50 m x 0.2 mm x 0.5 microm), then determined by IT-MS and quantified by external standard method. The plastic toy samples were extracted by the dissolution-precipitation approach, cleaned up with an Envi-carb solid phase extraction column and concentrated by rotary evaporation and nitrogen blowing, then determined by GC-IT-MS and quantified by external standard method. The calibration curves showed good linearity in the range of 0.002-50 mg/L with the correlation coefficients greater than 0.996 8. The limits of quantification (LOQ, S/N > 10) were 0.02-40 mg/kg. The average recoveries of the target compounds spiked in the sample at three concentration levels were in the range of 82.2%-110.8% with the relative standard deviations (RSDs) of 0.6%-10.5%. These results show that this method is accurate and sensitive for the qualitative and quantitative determination of the 21 fragrance allergens in the 3 types of toys.

Development and application of chemometric-assisted dispersive liquid-liquid microextraction for the determination of suspected fragrance allergens in water samples.

A simple and green method based on dispersive liquid-liquid microextraction, mated to chemometrics and followed by mass spectrometric detection for the determination of suspected fragrance allergens in water samples is developed and assessed in this work. Volume of extraction and disperser solvent, pH, ionic strength, extraction time, sample volume, as well as centrifugation time were initially optimized in a fractional factorial design. The obtained significant factors were optimized by using a central composite design and the quadratic model between the dependent and the independent variables was built. The obtained optimal conditions were: aqueous sample of 3.8 mL, 100 µL chloroform, 1.40 mL acetone, 4 min centrifugation time, natural pH containing 5% (w/v) NaCl, and centrifugation speed 4000 rpm. Method proved to be linear over a wide range of concentration for all analytes with R(2) between 0.9807 and 0.9959. The repeatability and reproducibility of the proposed method, expressed as relative standard deviation, varied between 3-13% and 4-16%, respectively. The limits of detection ranged from 0.007 to 1.0 µg L(-1) . The recommended method was applied to water samples including baby bath as well as swimming pool water samples and was compared with a previously reported method.

Membrane assisted solvent extraction coupled to large volume injection-gas chromatography-mass spectrometry for trace analysis of synthetic musks in environmental water samples.

This work describes the optimisation, validation and application of membrane assisted solvent extraction (MASE) together with a large volume injection (LVI) in a programmable temperature vaporisation (PTV) injector coupled to gas chromatography-mass spectrometry (GC-MS) for the quantification of ten synthetic musk fragrances (musks) in surface and wastewater samples. Regarding the MASE, musks were extracted from 150 mL of aqueous samples to 200 µL of n-hexane hold in home-made low density polyethylene (LDPE) bags. The extraction took 240 min and the performance of the method made possible the direct analysis of the extracts by LVI-PTV-GC-MS without needing any further treatment and avoiding losses of analytes. During the optimisation of LVI-PTV set-up, the response surfaces of every analyte signal against the cryo-focussing temperature, injection speed and vent time were built. Finally, the figures of merit of the whole procedure allowed the analysis of most of the musks owing to the low method detection limits (between 4 and 25 ng L⁻¹) and good precisions (<20%). In fact, this method was successfully applied to the analysis of musks in surface and wastewater samples. Galaxolide and tonalide are the main two synthetic musks observed in most of the analysed environmental water samples.

Comprehensive two-dimensional chromatography with coupling of reversed phase high performance liquid chromatography and supercritical fluid chromatography.

A 2D comprehensive chromatographic separation of blackberry sage fragrant oil was performed by using HPLC in the first dimension and SFC in the second. A C(18)-bonded silica column eluted with an ACN gradient was used in the HPLC dimension and an amino-bonded silica column eluted with ACN as a modifier in the SFC dimension. This 2D separation was completed in the off-line mode, the fractions from the HPLC column being collected and injected in the SFC column. The retention factors on the two columns have a -0.757 correlation coefficient. The method provides a practical peak capacity of 2400 in 280 min. The first eluted peaks in HPLC are the last ones eluted in SFC and vice versa. The results demonstrate that the coupling of an HPLC and an SFC separation have a great potential for 2D chromatographic separations.

A retention time locked gas chromatography-mass spectrometry method based on stir-bar sorptive extraction and thermal desorption for automated determination of synthetic musk fragrances in natural and wastewaters.

A stir-bar sorptive extraction (SBSE) method followed by automated thermal desorption (ATD) coupled to gas chromatography-mass spectrometry was optimized for determining trace levels of 18 synthetic fragrances (musks). Using the method developed a retention time locked library is created and converted to a screening database. This homebuilt database can be combined with deconvolution software for the identification of musks. A factorial design was provide to evaluate the main parameters and interactions between the factors affecting the process of SBSE. Operating with de MS-detector in the full-scan mode, high sensitivity with detection limits in the low ng L(-1) range, and good linearity and repeatability were achieved for all musks. The applicability of the method developed was tested in natural waters (surface and groundwater) and wastewater of a plant treatment (WWPT). The results obtained confirmed the usefulness of the proposed method for the determination and unequivocal identification of musks. This approach enables the developed method to be used for routine screening of environmental samples and posterior rapid quantitation of the positive samples.

Microemulsion electrokinetic chromatography: an application for the simultaneous determination of suspected fragrance allergens in rinse-off products.

A mixture of 18 neutral UV-active compounds with different characteristics of polarity was determined by capillary electrophoresis using a pseudostationary phase constituted by a microemulsion. The test analytes were volatile fragrance compounds, included in a list of 24 chemicals classified as suspected allergens according to Directive 2003/15/CE. The considered compounds were detected at 195 nm and p-anisaldehyde was chosen as internal standard. The background electrolyte consisted of a standard microemulsion made of 90.95% 10mM borax buffer, pH 9.2, 1.05% n-heptane, 8.00% SDS/n-butanol in 1:2 ratio, to which 40 mM methyl-ß-cyclodextrin was added. Temperature and voltage were set at 20 °C and 25 kV, respectively. These experimental conditions allowed separation of the compounds to be obtained in about 20 min. The method was applied to real samples made up of rinse-off scented products. The results obtained using the standard microemulsion as pseudostationary phase showed its high resolution power, capable of effectively separating a complex mixture of analytes. Microemulsion electrokinetic chromatography was confirmed to have a great potential for different analytical challenges, holding up the possibility of using this technique as a good and complementary alternative to HPLC methods for routine analysis.

Ultrasound-assisted emulsification-microextraction of fragrance allergens in water.

A method based on ultrasound-assisted emulsification-microextraction (USAEME) and gas chromatography-mass spectrometry (GC-MS) has been developed for the analysis of regulated fragrance allergens in water. Extraction conditions such as the type of solvent, extraction temperature, irradiation time, and salting-out effect were optimized using a multivariate approach. Compounds were extracted during 2 min in an acoustically emulsified media formed by 100 µL chloroform and 10 mL sample. The USAEME process provided an efficient and exhaustive extraction (enrichment factor ∼100) and, after centrifugation, the extract was ready for GC analysis. Validation was performed using spiked ultrapure water as well as other most complex matrices such as sewage water. Recoveries between 75% and 110% were generally obtained, and precision was characterized by RSD values <10% in most cases. The limits of detection (LODs) were at the sub-nanogram per millilitre level. The proposed procedure was applied to the determination of allergens in several real samples including tap water, baby bathwater, recreational place water, public washing place water, and sewage water. The presence of some of the target compounds was confirmed in all the samples excluding tap water, demonstrating the ubiquity of this group of cosmetic and personal care products ingredients.

Volatiles from leaves and rhizomes of Fragrant Acorus spp. (Acoraceae).

Three horticultural selections of Acorus gramineus Soland were investigated to determine the chemical composition of their leaves and rhizomes. The variety 'liquorice' was found to contain methylchavicol (49%) which accounts for the unusual anisic odor of this variety, while beta-asarone was the main component of A. christophii (43%) and 'yodo-no-yuki' (20%). The results are compared with calamus oils, and the possible biosynthetic precursors of the main components methylchavicol and beta-asarone are considered.