Synthesis and characterization of polysaccharide- and protein-based edible films and application as packaging materials for fresh fish fillets.

The rising packaging industry together with global demand for sustainable production has increased the interest in developing biodegradable packaging materials. The aim of the study was to develop edible films based on pectin, gelatin, and hydroxypropyl methylcellulose and evaluate their applicability as biodegradable packaging materials for gilthead seabream fillets. Mechanical properties, water barriers, wettability of the films through contact angle measurement, optical, and UV-Vis barrier properties were evaluated for food packaging applications. The effective blend of polysaccharide and protein film-forming solutions was confirmed by the produced films with excellent optical properties, acceptable mechanical properties and adequate barriers to water vapor. The contact angle for pectin based and gelatin based films were higher than 90° indicating the hydrophobic films, while HPMC based films had contact angle lower than 90°. The produced films were tested as alternative and environmentally friendly packaging materials for gilthead seabream fillets during refrigerated storage. All tested packaging conditions resulted in similar shelf-life in packed gilthead seabream fillets (i.e. 7-8 days at 2 °C). The results showed that the developed films may reduce the use of conventional petroleum-based food packaging materials without affecting the shelf-life of fish.

Study of volatile compounds in Greek pistachio (Pistacia vera L. 'Aegina' cultivar) oils using Soxhlet and ultrasound assisted extraction.

Headspace solid-phase microextraction followed by gas chromatography-mass spectrometry (HS-SPME/GC-MS) represents the most used solvent-free methodology for the characterization of the complex and heterogeneous mix of volatile compounds. The present study investigates the differences in volatile profile of pistachio oils 'Aegina' cultivar extracted with two different techniques, ultrasound assisted extraction (UAE) and Soxhlet. Differences were observed both in the pistachio oil yield and the composition of the volatile compounds among these two groups of samples, which were significantly influenced due to the different thermal conditions. In terms of pistachio oil yield, the Soxhlet extraction technique was proven more efficient (52.5-68.2% w/w) than the UAE type (28.2-42.6% w/w). A total of 34 and 30 volatile compounds were identified for UAE and Soxhlet, respectively. The main ones associated with UAE were α-pinene, octane and decane, while the volatiles formed as a consequence of Soxhlet extraction were decane, nonanal and (E)-2-decenal. Terpenes' concentrations were found decreased in Soxhlet's samples, but hydrocarbons and aldehydes were significantly increased in these samples. Numerous studies concluded in common results. However, this article is the first to explore the influence of different extraction types on the volatile profile of the unique flavour and odor pistachio oil 'Aegina' cultivar.

The application of right-angle fluorescence spectroscopy as a tool to distinguish five autochthonous commercial Greek white wines.

White wine is among the most widely consumed alcoholic beverages. Varietal discrimination of wines has received increasing attention. Today's consumers require a sense of authenticity and are deterred by falsehood or misrepresentation in product marketing. However, wine can involve various types of frauds, which directly affects the distribution of wine in national and international markets. Right-angle fluorescence spectroscopy is a simple and rapid analytical technique that in combination with chemometric algorithms, constitutes a novel method for wine authentication. In this study, the stepwise-Linear Discriminant Analysis algorithm was applied in three representative spectral regions related to phenolic compounds for the purpose of distinguishing white wines according to the grape variety. The wavelength at 310 nm attributed to the hydroxycinnamic acids and stilbene provided a higher classification rate (95.5%) than the λex 280 and 295 nm regions (79.8%), suggesting that these compounds are highly related to the botanical origin of samples. The chemometric models were validated utilizing cross-validation and an external validation set to enhance the robustness of the proposed methodology. The above-mentioned methodology constitutes a powerful tool for the varietal discrimination of white wines and can be used in industrial setting. The ultimate goal of this study is to contribute to the efforts towards the authentication of Greek white wine which will eventually support producers and suppliers to remain competitive and simultaneously protect the consumers from fraudulent practices.

Unifloral Autumn Heather Honey from Indigenous Greek Erica manipuliflora Salisb.: SPME/GC-MS Characterization of the Volatile Fraction and Optimization of the Isolation Parameters.

For long heather honey has been a special variety due to its unique organoleptic characteristics. This study aimed to characterize and optimize the isolation of the dominant volatile fraction of Greek autumn heather honey using solid-phase microextraction (SPME) followed by gas chromatography-mass spectrometry (GC-MS). The described approach pointed out 13 main volatile components more closely related to honey botanical origin, in terms of occurrence and relative abundance. These volatiles include phenolic compounds and norisoprenoids, with benzaldehyde, safranal and p-anisaldehyde present in higher amounts, while ethyl 4-methoxybenzoate is reported for the first time in honey. Then, an experimental design was developed based on five numeric factors and one categorical factor and evaluated the optimum conditions (temperature: 60 °C, equilibration time: 30 min extraction time: 15 min magnetic stirrer velocity: 100 rpm sample volume: 6 mL water: honey ratio: 1:3 (v/w)). Additionally, a validation test set reinforces the above methodology investigation. Honey is very complex and variable with respect to its volatile components given the high diversity of the floral source. As a result, customizing the isolation parameters for each honey is a good approach for streamlining the isolation volatile compounds. This study could provide a good basis for future recognition of monofloral autumn heather honey.

GC-MS, FTIR and Raman spectroscopic analysis of fatty acids of Pistacia vera (Greek variety "Aegina") oils from two consecutive harvest periods and chemometric differentiation of oils quality.

Pistacia vera oil is a rich source of unsaturated fatty acids, whose presence is associated with high quality and nutritional value. According to the literature, fatty acid oil composition is not constant every harvest year, but varies mainly depending on climate conditions. Therefore, the knowledge of oil composition in fatty acids is necessary to assess both its quality and its nutritional value. Twenty-two samples (11 samples from the harvest year 2017 and 11 samples from 2018) of the Greek variety "Aegina" were collected from four different Greek regions, from producers following the same cultivation and post-harvest cares. Extraction oil yields were found to be similar (61.7% w/w, 2017; 60.8% w/w, 2018). A reduction of the saturated fatty acids content was determined in 2018 (mean values 12.2% w/w against 13.8% w/w in 2017) by Gas Chromatography-Mass Spectrometry, accompanied by an increase of the unsaturated ones (mean values 87.9% w/w against 86.2% w/w in 2017). These results indicate that the harvest year 2018 may be considered superior to 2017 in terms of quality and nutritional value and may be correlated with an increased mean rain rate in 2018 and a slight decrease of the mean temperature. Fourier transform infrared (FTIR) and Raman spectroscopic studies of the oils were also performed. Three chemometric models were developed for the two consecutive harvest years of pistachio oil and the discrimination was based on GC-MS analysis, FTIR and Raman spectroscopic data combined with cross-validation techniques and comparison among them. The most successful chemometric model was that based on FTIR spectroscopy, which has the advantage of speed, simplicity and economy. Such a chemometric model may help in estimating the quality of Pistacia vera oils.

Authentication of the Botanical and Geographical Origin and Detection of Adulteration of Olive Oil Using Gas Chromatography, Infrared and Raman Spectroscopy Techniques: A Review.

Olive oil is among the most popular supplements of the Mediterranean diet due to its high nutritional value. However, at the same time, because of economical purposes, it is also one of the products most subjected to adulteration. As a result, authenticity is an important issue of concern among authorities. Many analytical techniques, able to detect adulteration of olive oil, to identify its geographical and botanical origin and consequently guarantee its quality and authenticity, have been developed. This review paper discusses the use of infrared and Raman spectroscopy as candidate tools to examine the authenticity of olive oils. It also considers the volatile fraction as a marker to distinguish between different varieties and adulterated olive oils, using SPME combined with gas chromatography technique.

Composition, seasonal abundance, and public health importance of mosquito species in the regional unit of Thessaloniki, Northern Greece.

Mosquitoes (Diptera: Culicidae) are the largest group of blood-feeding insects that disturb not only humans but also other mammals and birds. This study reports the presence of native mosquito species in the regional unit of Thessaloniki and the monitoring of their population. In total, 13 mosquito species belonging to four genera were identified. The most dominant species was Culex pipiens, followed by Aedes caspius. In the present study, we report for the first time the presence of Ae. vittatus in Greece and of Anopheles plumbeus in the regional unit of Thessaloniki. Regarding the seasonal variation, species of the genus Aedes were the ones that first appeared in late March, followed by Culex species at the end of April and finally species of the genus Anopheles in July. Species of the Aedes genus were found to be the most abundant in the first quarter of the year (late March to early April). Population of Cx. pipiens remained at high levels from late April to late September. Species of the genus Anopheles were found in high densities from early August to October. The current study contributes to the knowledge of the mosquito species composition and their relative abundance in an area where West Nile virus caused severe epidemic outbreaks.

Response Surface Methodology to Optimize the Isolation of Dominant Volatile Compounds from Monofloral Greek Thyme Honey Using SPME-GC-MS.

This study aimed at an experimental design of response surface methodology (RSM) in the optimization of the dominant volatile fraction of Greek thyme honey using solid-phase microextraction (SPME) and analyzed by gas chromatography-mass spectrometry (GC-MS). For this purpose, a multiple response optimization was employed using desirability functions, which demand a search for optimal conditions for a set of responses simultaneously. A test set of eighty thyme honey samples were analyzed under the optimum conditions for validation of the proposed model. The optimized combination of isolation conditions was the temperature (60 °C), equilibration time (15 min), extraction time (30 min), magnetic stirrer speed (700 rpm), sample volume (6 mL), water: honey ratio (1:3 v/w) with total desirability over 0.50. It was found that the magnetic stirrer speed, which has not been evaluated before, had a positive effect, especially in combination with other factors. The above-developed methodology proved to be effective in the optimization of isolation of specific volatile compounds from a difficult matrix, like honey. This study could be a good basis for the development of novel RSM for other monofloral honey samples.

Chemometric Study of Fatty Acid Composition of Virgin Olive Oil from Four Widespread Greek Cultivars.

Virgin olive oil (VOO) is one of the key components of the Mediterranean diet owing to the presence of monounsaturated fatty acids and various bioactive compounds. These beneficial traits, which are usually associated with the cultivar genotype, are highlighting the demand of identifying characteristics of olive oil that will ensure its authenticity. In this work, the fatty acid (FA) composition of 199 VOO samples from Koroneiki, Megaritiki, Amfissis, and Manaki cultivars was determined and studied by chemometrics. Olive cultivar greatly influenced the FA composition, namely, oleic acid (from 75.36% for Amfissis to 65.81% for Megaritiki) and linoleic acid (from 13.35% for Manaki to 6.70% for Koroneiki). Spearman's rho correlation coefficients revealed differences and similarities among the olive oil cultivars. The use of the forward stepwise algorithm identified the FAs arachidonic acid, gadoleic acid, linoleic acid, α-linolenic acid, palmitoleic acid, and palmitic acid as the most significant for the differentiation of samples. The application of linear and quadratic cross-validation discriminant analysis resulted in the correct classification of 100.00% and 99.37% of samples, respectively. The findings demonstrated the special characteristics of the VOO samples derived from the four cultivars and their successful botanical differentiation based on FA composition.

Rapid screening on aflatoxins' presence in Pistachia vera nuts using diffuse reflectance infrared Fourier transform spectroscopy and chemometrics.

Aflatoxin contamination in pistachios has been analyzed in this work, using Diffuse Reflectance Infrared Spectroscopy (DRIFTS) with chemometrics. Forty-nine Greek pistachio samples of different aflatoxin concentrations were classified into aflatoxin and non-aflatoxin groups using the 3035-2821, 1770-1721, 1570-1481 and 1260-1061 cm-1 spectral regions in Kubelka-Munk conversion and first derivative form. A chemometric model was developed using twenty-eight samples as calibration, 11 as validation and 10 as test set. The discrimination analysis separated correctly the 100% of the calibration and the validation set and the 80% of the test set. The proposed chemometric model is simple, rapid, economical and environmentally friendly since it does not require chemical pre-treatment of the samples. It is expected that the present method may be proved useful in food industry and the inspection authorities as a rapid decision-making tool to detect batches that must be rejected and enhance consumers' protection from aflatoxin contaminated pistachios.

Discrimination of botanical origin of olive oil from selected Greek cultivars by SPME-GC-MS and ATR-FTIR spectroscopy combined with chemometrics.

BACKGROUND: Consumers today wish to know the botanical origin of the olive oil they purchase. The objective of the present study was the development of robust chemometric models based on gas chromatography-mass spectrometry (GC-MS) and attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) for the purpose of botanical differentiation of three commercial Greek olive oil cultivars. RESULTS: Using the solid-phase microextraction technique (SPME), volatile compounds (VC) were obtained and analyzed by GC-MS. Five hydrocarbons and one ester were selected by the forward stepwise algorithm, which best discriminated the olive oil samples. From ATR-FTIR analysis, the spectral regions chosen from the forward stepwise algorithm were associated with CO stretching vibration of the esters of triglycerides and the CH bending vibrations of the CH2 aliphatic group and double bonds. Application of the supervised methods of linear and quadratic discriminant cross-validation analysis, based on VC data, provided a correct classification score of 97.4% and 100.0%, respectively. Corresponding statistical analyses were used in the mid-infrared spectra, by which 96.1% of samples were discriminated correctly. CONCLUSION: ATR-FTIR and SPME-GC-MS techniques in conjunction with the appropriate feature selection algorithm and classification methods proved to be powerful tools for the authentication of Greek olive oil. The proposed methodology could be used in an industrial setting for determination of the botanical origin of Greek olive oil. © 2020 Society of Chemical Industry.

Botanical origin discrimination of Greek honeys: physicochemical parameters versus Raman spectroscopy.

BACKGROUND: The authenticity of honey is of high importance since it affects its commercial value. The discrimination of the origin of honey is of prime importance to reinforce consumer trust. In this study, four chemometric models were developed based on the physicochemical parameters according to European and Greek legislation and one using Raman spectroscopy to discriminate Greek honey samples from three commercial monofloral botanical sources. RESULTS: The results of physicochemical (glucose, fructose, electrical activity) parameters chemometric models showed that the percentage of correct recognition fluctuated from 92.2% to 93.8% with cross-validation 90.6-92.2%, and the placement of test set was 79.0-84.3% successful. The addition of maltose content in the previous discrimination models did not significantly improve the discrimination. The corresponding percentages of the Raman chemometric model were 95.3%, 90.6%, and 84.3%. CONCLUSION: The five chemometric models developed presented similar and very satisfactory results. Given that the recording of Raman spectra is simple, fast, a minimal amount of sample is needed for the analysis, no solvent (environmentally friendly) is used, and no specialized personnel are required, we conclude that the chemometric model based on Raman spectroscopy is an efficient tool to discriminate the botanical origin of fir, pine, and thyme honey varieties. © 2020 Society of Chemical Industry.

Study of the Quality Parameters and the Antioxidant Capacity for the FTIR-Chemometric Differentiation of Pistacia Vera Oils.

The aim of this work was to characterize the pistachio oil of the Greek variety, "Aegina", evaluate its various quality indices, and investigate the potential use of FTIR as a tool to discriminate different oil qualities. For this purpose, the antioxidant capacity, the tocopherol content and the oxidation and degradation of fatty acids, as described by k, Δk, R-values, and free acidity were evaluated using 45 samples from eight different areas of production and two subsequent years of harvesting. The antioxidant capacity was estimated using 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid diammonium salt (ABTS) and 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazine (DPPH) assays, and the tocopherol content was quantified through HPLC analysis. FTIR spectra were recorded for all samples and multivariate analysis was applied. The results showed significant differences between the oil samples of different harvesting years, which were successfully discriminated by a representative FTIR spectral region based on R-value, total antioxidant capacity, and scavenging capacity, through ABTS. A similar approach could not be confirmed for the other quality parameters, such as the free acidity and the tocopherol content. This research highlighted the possibility of developing a simple, rapid, economic, and environment friendly method for the discrimination of pistachio oils according to their quality profile, through FTIR spectroscopy and multivariate analysis.

Differentiation and identification of grape-associated black aspergilli using Fourier transform infrared (FT-IR) spectroscopic analysis of mycelia.

The purpose of this study was to evaluate the potential of FT-IR spectroscopy as a high-throughput method for rapid differentiation among the ochratoxigenic species of Aspergillus carbonarius and the non-ochratoxigenic or low toxigenic species of Aspergillus niger aggregate, namely A. tubingensis and A. niger isolated previously from grapes of Greek vineyards. A total of 182 isolates of A. carbonarius, A. tubingensis, and A. niger were analyzed using FT-IR spectroscopy. The first derivative of specific spectral regions (3002-2801cm-1, 1773-1550cm-1, and 1286-952cm-1) were chosen and evaluated with respect to absorbance values. The average spectra of 130 fungal isolates were used for model calibration based on Discriminant analysis and the remaining 52 spectra were used for external model validation. This methodology was able to differentiate correctly 98.8% in total accuracy in both model calibration and validation. The per class accuracy for A. carbonarius was 95.3% and 100% for model calibration and validation, respectively, whereas for A. niger aggregate the per class accuracy amounted to 100% in both cases. The obtained results indicated that FT-IR could become a promising, fast, reliable and low-cost tool for the discrimination and differentiation of closely related fungal species.

Comparative evaluation of an ISO 3632 method and an HPLC-DAD method for safranal quantity determination in saffron.

The aim of this work was a comparison of the ISO 3632 (2011) method and an HPLC-DAD method for safranal quantity determination in saffron. Samples from different origins were analysed by UV-vis according to ISO 3632 (2011) and by HPLC-DAD. Both methods were compared, and there was no correlation between the safranal content obtained by UV-vis and HPLC-DAD. An over-estimation in the UV-vis experiment was observed, which was related to the cis-crocetin esters content, as well as other compounds. The results demonstrated that there was no relationship between ISO quality categories and safranal content using HPLC-DAD. Therefore, HPLC-DAD might be preferable to UV-vis for determining the safranal content and the classification of saffron for commercial purposes. In addition, HPLC-DAD was adequate for determining the three foremost parameters that define the quality of saffron (crocetin esters, picrocrocin and safranal); therefore, this approach could be included in the ISO 3632 method (2011).

Impact of fluctuating temperatures on development of the koinobiont endoparasitoid Venturia canescens.

The effect of temperature on the biology of Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae) is well understood under constant temperature conditions, but less so under more natural, fluctuating conditions. Herein we studied the influence of fluctuating temperatures on biological parameters of V. canescens. Parasitized fifth-instar larvae of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) were reared individually in incubators at six fluctuating temperature regimes (15-19.5°C with a mean of 17.6°C, 17.5-22.5°C with a mean of 19.8°C, 20-30°C with a mean of 22.7°C, 22.5-27.5°C with a mean of 25°C, 25.5-32.5°C with a mean of 28.3°C and 28.5-33°C with a mean of 30°C) until emergence and death of V. canescens adults. Developmental time from parasitism to adult eclosion, adult longevity and survival were recorded at each fluctuating temperature regime. In principle, developmental time decreased with an increase of the mean temperature of the fluctuating temperature regime. Upper and lower threshold temperatures for total development were estimated at 34.9 and 6.7°C, respectively. Optimum temperature for development and thermal constant were 28.6°C and 526.3 degree days, respectively. Adult longevity was also affected by fluctuating temperature, as it was significantly reduced at the highest mean temperature (7.0 days at 30°C) compared to the lowest one (29.4 days at 17.6°C). Survival was low at all tested fluctuating temperatures, apart from mean fluctuating temperature of 25°C (37%). Understanding the thermal biology of V. canescens under more natural conditions is of critical importance in applied contexts. Thus, predictions of biological responses to fluctuating temperatures may be used in population forecasting models which potentially influence decision-making in IPM programs.

Direct determination of rosmarinic acid in Lamiaceae herbs using diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) and chemometrics.

For the determination of rosmarinic acid (RA) directly in pulverized plant material, a method is developed using diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) without any physicochemical pretreatment of samples. The RA content of 11 samples of eight different Lamiaceae herbs, as determined by high performance liquid chromatography (HPLC), varied between 86 ± 1 mg/g (in lemon balm) and 12.0 ± 0.8 mg/g (in hyssop) of dried plant material. The 11 samples and 50 other additional samples, which were prepared by mixing initial samples with KBr, were measured using DRIFTS. The second derivative of the spectral region 1344-806 cm(-1) was used and the corresponding data were analyzed using partial least squares (PLS) regression. The correlation between infrared spectral analysis and HPLC measurements shows that the DRIFTS method is sufficiently accurate, simple, and rapid. The RA content of the 11 Lamiaceae samples determined by DRIFTS ranged from 81 ± 4 mg/g (in lemon balm) to 12 ± 3 mg/g (in hyssop) of dried plant material.

Rapid strain classification and taxa delimitation within the edible mushroom genus Pleurotus through the use of diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy.

Fourier transform infrared (FT-IR) spectroscopy has been successfully applied for the identification of bacteria and yeasts, but only to a limited extent for discriminating specific groups of filamentous fungi. In the frame of this study, 73 strains - from different associated hosts/substrates and geographic regions - representing 16 taxa of the edible mushroom genus Pleurotus (Basidiomycota, Agaricales) were examined through the use of diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy. A binary matrix, elaborated on the basis of presence/absence of specific absorbance peaks combined with cluster analysis, demonstrated that the spectral region 1800-600 cm(-1) permitted clear delimitation of individual strains into Pleurotus species. In addition, closely related species (e.g., Pleurotus ostreatus and Pleurotus pulmonarius) or taxa of the subgenus Coremiopleurotus demonstrated high similarity in their absorbance patterns, whereas genetically distinct entities such as Pleurotus dryinus, Pleurotus djamor, and Pleurotus eryngii provided spectra with noteworthy differences. When specific regions (1800-1700, 1360-1285, 1125-1068, and 950-650 cm(-1)) were evaluated in respect to the absorbance values demonstrated by individual strains, it was evidenced that this methodology could be eventually exploited for the identification of unknown Pleurotus specimens with a stepwise process and with the aid of a dichotomous key developed for this purpose. Moreover, it was shown that the nature of original fungal material examined (mycelium, basidiomata, and basidiospores) had an effect on the outcome of such analyses, and so did the use of different mycelium growth substrates. In conclusion, application of FT-IR spectroscopy provided a fast, reliable, and cost-efficient solution for the classification of pure cultures from closely related mushroom species.

Quantitative determination of pulegone in pennyroyal oil by FT-IR spectroscopy.

Pulegone constitutes a monoterpene occurring in Mentha species and primarily in Mentha pulegium L. (pennyroyal). A major source of human exposure to pulegone is the use of pennyroyal essential oil in flavorings, confectionery and cosmetics. The rapid quantification of pulegone in hydrodistilled pennyroyal oils (which were also "spiked" to increase the validation range) by Fourier transform infrared spectroscopy (FT-IR) combined with partial least-squares (PLS) regression was evaluated, using the spectral region 1650-1260 cm(-1). Gas chromatography was applied as the reference method for pennyroyal oil samples, which ranged in pulegone content from 157 to 860 mg/mL. The two methods were subjected to statistical tests and proved equivalent in terms of accuracy and reproducibility (99% confidence level). The use of FT-IR spectroscopy could offer a viable alternative to the standard analysis procedures presently applied for quantification of valuable plant substances and could also provide the processing industry with a simple and high-throughput technique for the fast quality check of incoming raw materials such as pennyroyal oils.

Comparison of distillation and ultrasound-assisted extraction methods for the isolation of sensitive aroma compounds from garlic (Allium sativum).

A comparative study of traditional simultaneous distillation extraction (SDE), microwave assisted hydrodistillation extraction (MWHD) and ultrasound-assisted extraction (USE) is presented, for the extraction of essential oils from fresh garlic (Allium sativum) cloves. Each method is evaluated in terms of qualitative and quantitative composition of the isolated essential oil. The highly reactive sulfur molecules of the garlic volatile fraction show variable response to the different isolation methods. The application of ultrasound for the extraction of the essential oil is considered to cause a lesser damage of thermal-sensitive molecules, thus, providing a better approach of the compounds primarily responsible for the characteristic odor and taste of freshly chopped garlic. All heat-involving isolation procedures have been shown to differentiate the volatile-fraction profile as analyzed by GC-MS. Especially when grouping the compounds into cyclic and acyclic, the percentage concentrations drop from 77.4% to 8.7% for the acyclic while that of the cyclic compounds increase from 4.7% to 70.8%. The observed fact may be attributed to the effect of the heat applied, which changes from harsh thermal treatment (SDE) to short time thermal (MWHD) and room-temperature isolation (USE). The use of USE proves to be crucial in order to provide reliable insight into garlic's chemistry.