A review of volatile analytical methods for determining the botanical origin of honey

Aroma is an important quality factor in foods. The aroma of bee honey depends on volatile fraction composition, which is influenced by nectar composition and floral origin. Honey of unifloral origin usually commands higher commercial value, thus the floral determination and certification of unifloral honey plays an important role in quality control. This review concerns investigations made on the volatile fraction of bee honey by gas chromatography/mass spectrometry. Recent advances in extraction methods, results achieved, and comparisons of alternative dependable methods for determining floral origin of bee honey are discussed. We emphasize solid phase micro-extraction gas chromatography (SPME/GC) methodology and present some of the results obtained to date, plus the advantages and drawbacks of SPME/GS in comparison with other methods.     

Some aspects of dynamic headspace analysis of volatile components in honey

Purge and Trap (P&T) has been evaluated as a fractionation technique for the GC–MS analysis of the most volatile compounds in honey. The volatile fraction of twenty-two commercial honeys of eight different botanical sources (eucalyptus, thyme, citrus, rosemary, heather, lavender, multiflower and honeydew) was characterized by P&T–GC–MS. Hundred volatile compounds were characterized, 18 of them being determined for the first time in honey. Compounds detected included volatiles derived from the floral nectar or honeydew source such as terpenes, furan derivatives from honey processing and storage and other compounds whose origin could be related to microbial or environmental contamination. This method has also been validated with regard to data reproducibility; with relative data (percentage of total volatile composition) showing a better precision over quantitative results calculated using an internal standard. Application of multivariate statistical analysis to P&T–GC–MS data has shown to be very promising to classify samples according to their botanical origin.     

Effect of pork fat addition on the volatile compounds of foal dry-cured sausage

The effect of fat content on volatile compounds from foal dry-cured sausage was studied. Three batches (10 units per batch) of dry fermented sausages with different pork back fat content (5%, 10% and 20%) were manufactured; low fat (LF), medium fat (MF) and high fat (HF), respectively. A total of 45 volatile compounds were extracted by purge-and-trap and identified by GC-MS in the headspace of the batches. The mixture comprised 11 terpenes, 15 esters, 14 hydrocarbons and 2 alcohols. Spices were responsible for the generation of 14 volatile compounds comprising terpenes and hydrocarbons. After 49 days of ripening volatile compounds from spices represented 52.9, 38.9 and 31.2% of the total area for samples from LF, MF and HF batches, respectively, while lipid autooxidation was responsible for the generation of 8 volatile compounds. The PCA offered a good separation of the mean samples according to their fat content.     

Contribution of enterococci to the volatile profile of slightly-fermented sausages

The volatile profiles of slightly-fermented sausages inoculated with eight different enterococci strains were determined and compared with a fermented sausage manufactured without any inoculation. The volatile profile was determined by solid-phase microextraction (SPME) and gas chromatography coupled to mass spectrometry (GC/MS). A total of 121 volatile and semi-volatile compounds were identified in the samples of the final products (day 21). Terpenes from spices added during manufacture of dry-fermented sausages were the main group of compounds in the volatile profile. Other groups included alcohols (18% of total volatile compounds), aldehydes (16%), alkanes and alkenes (12%), and ketones (10%). These are derived from lipids, proteins or carbohydrates and formed during fermentation and ripening. In terms of global volatile composition, the batches inoculated with enterococci strains were similar to those obtained for non-inoculated batch. However, some quantitative differences in the volatile profiles of different batches were found, mainly in the compounds formed from amino acid catabolism (i.e.: propanol, isoamyl), from free fatty acid oxidation (i.e.: hexanal, hexanol), or from carbohydrate fermentation (i.e.: 2-butanone, acetoin). In comparison with non-inoculated sausages, some batches were more resistant to oxidation; they contained fewer volatile compounds arising from lipid oxidation. The use of different selected enterococci strains did not alter the volatile profile of slightly-fermented sausages. The enterococci strains provide similar volatile compounds to those usually reported in the literature.     

GC/MS法分析烟用接装纸中挥发性有机化合物

建立了顶空-气质联用(HS-GC/MS)法,对烟用接装纸中挥发性有机化合物(VOCs)残留量进行测试.结果表明:(1)该方法检测限为0.001-0.004 mg/mL,回收率为89.35％- 108.96％,相对标准偏差(RSD)＜4％;(2)所有52个被检烟用接装纸榉品均为合格样品,所有样品中均未检出苯、乙苯、二甲苯、乙酸异丙酯、丁酮,YC171-2009《烟用接装纸》标准中规定的5种VOCs成分中只有乙酸正丁酯有个别检出;(3)醇类检出率较高,其中所有样品都有乙醇检出,异丙醇、正丁醇的检出率也比较高;(4)丙二醇甲醚有较高的检出率,且其检出量相对较大.研究结果充分说明,该方法适合于烟用接装纸中VOCs的快速批量检测.     

A potentiometric electronic tongue for the discrimination of honey according to the botanical origin. Comparison with traditional methodologies: Physicochemical parameters and volatile profile

The effectiveness of a potentiometric electronic tongue, made of various metals and metallic compounds was evaluated for the differentiation of honey in three different states: raw, liquefied and pasteurized. Principal component analysis (PCA) and a neural network showed that potentiometric electrodes seem useful to classify honey by its botanical origin, though they do not seem capable of discrimination between applied thermal treatments. The same was observed in the samples for physicochemical parameters and volatile compounds analysed with PCA. Au, Cu and Ag electrodes were more decisive in the discrimination of honey. A remarkable correlation (PLS analysis) between the electronic tongue and the physicochemical parameters was found, the best results being for color Pfund (r² = 0.958), L^∗ (r² = 0.935) and diastase activity (r² = 0.926). The correlation with volatile compounds was much weaker, though this improved for specific characteristic compounds in each monofloral variety. Thus, this is a good starting point for the development of new systems focused on the honey sector.     

Botanical discrimination and classification of honey samples applying gas chromatography/mass spectrometry fingerprinting of headspace volatile compounds

A validated method for the discrimination and classification of honey samples performing GC/MS fingerprinting of headspace volatile compounds was developed. Combined mass spectra of honey samples originated from different plants and geographical regions of Greece were subjected to orthogonal partial least squares-discriminant analysis™ (OPLS™-DA), soft independent modelling of class analogy (SIMCA), and OPLS™-hierarchical cluster analysis (OPLS™-HCA). Analyses revealed an excellent separation between honey samples according to their botanical origin with the percentage of misclassification to be as low as 1.3% applying OPLS™-HCA. Fragments (m/z) responsible for the observed separation were assigned to phenolic, terpenoid, and aliphatic compounds present in the headspace of unifloral honeys. On the other hand, a variable classification for citrus and thyme honeys according to their geographical origin could be achieved. Results suggested that the developed methodology is robust and reliable for the botanical classification of honey samples, and the study of differences in their chemical composition.     

Influence of simulated industrial thermal treatments on the volatile fractions of different varieties of honey

The aim of this study was to determine if the volatile fraction of honey is affected by the application of standard industrial thermal treatment processes. Four types of Spanish honey were studied: three of floral origin (citrus, rosemary and polyfloral) and the fourth from honeydew. Each sample of honey was divided into three parts: one was left untreated, one was liquefied (at 45 °C for 48 h) and the other was both liquefied and pasteurized (at 80 °C for 4 min). All the samples analyzed were characterized to determine their melissopalynological, physicochemical (pH, moisture, total acidity, conductivity, hydroxymethylfurfural, and diastase activity), and volatile profiles. Type of honey had a greater impact on volatile fraction variations than did heat treatment. The overall volatile profile of each kind of honey permitted the classification of the honeys by botanical origin, revealing that there were practically no differences between the raw, liquefied, and pasteurized samples of each honey. These findings suggest that industrial processes conducted under controlled conditions should not significantly alter the intrinsic aroma of honey.     

Effect of high-pressure processing on volatile composition and odour of cherry tomato purée

The impact of high-pressure processing on the odour of cherry tomato purée was evaluated by sensory evaluation and SPME–GC/MS analysis. Two temperatures (20 and 60 °C) and two pressures (atmospheric and 800 MPa) were used in processing. Higher pressure at ambient temperature decreased the levels of certain volatile aldehydes, ketones and alcohols present in tomato, whereas, the levels of hexanal, heptanal and octanal increased. The higher temperature combined with either ambient or high-pressure, decreased the levels of many volatile compounds and caused a reduction in the intensity of fresh tomato odour. Processing at 800 MPa and 60 °C resulted in a marked increase in the intensity of cooked tomato and tea-like odour. On the basis of sensory assessment and volatile analysis, high pressure treatment at 800 MPa does not seem to be suitable for preserving fresh tomato odour.     

Determination of fermentative volatile compounds in aged red wines by near infrared spectroscopy

The purpose of this work was to study the possibility of determination of fermentative volatile compounds in aged red wines using NIR spectroscopy. To achieve this, 240 wines belonging to different geographic zones and elaborated with one or two varieties were analyzed. The volatile compounds were quantified by SBSE-GC–MS. Spectra obtained by NIR were co-related with these values using partial least square (PLS) regression. Calibration and validation statistics showed the quality of the model, after all when is done separately for each of the four geographic zones, and in the case of wines elaborated with two varieties. Consequently, near infrared spectroscopy can be used as an easy and rapid tool to determine fermentative volatile compounds in aged red wines.     

Characterisation of natural honey proteins: implications for the floral and geographical origin of honey

Characterisation of honey proteins has been considered advantageous for differentiating floral and geographical origin of honey. We analysed protein profiles of multi‐ and mono‐floral honey samples from different regions and harvest dates. The molecular masses of chromatographic peaks were in the range of 10–>200 kDa. Owing to comparable molecular masses, interesting correlation between profiles of honey proteins reported earlier and observed in this study could be established. Chromatograms of honey proteins revealed three novel peaks of molecular masses ～220, 129 and 26 kDa. Statistical analysis of peak areas showed that the 84‐kDa peak was different among all honey samples and the 220‐kDa peak was important for differentiation between multi‐ and mono‐floral honey samples. Chromatograms of several‐year‐old honey samples were different from fresh samples because of depletion of high molecular mass peaks that indicated in situ proteolysis. The results supported the notion of applying gel filtration as cost‐effective and robust technique for honey protein characterisation.     

Effect of the inclusion of chestnut in the finishing diet on volatile compounds during the manufacture of dry-cured “Lacón” from Celta pig breed

The effect of the finishing diet on the volatile compounds throughout the manufacture of dry-cured “lacón” (a Spanish traditional meat product), from the Celta pig breed was studied. Thirty-six pigs were separated into three groups according to the type of feeding during the finish-fattening period of three months (concentrate, mixed diet and chestnut). From the pigs of each diet, four batches of dry-cured “lacón” were manufactured. From each batch, samples of fresh meat, meat after salting, after post-salting, and after 14, 28, 56 and 84 days of drying–ripening were taken. Volatiles were extracted by a purge-and-trap method and analyzed by gas chromatographic/mass spectrometry (GC/MS). Seventy-six volatile compounds were identified and quantified from dry-cured “lacón” samples in pigs finished with chestnut, eighty-two for concentrate fed pigs and eighty in pigs fed with the mixed diet. The number of identified volatile compounds increased during the manufacturing process; at 84 days of drying–ripening, in the dry-cured “lacón” samples from pigs finished with concentrate, mixed diet and chestnut, 54, 58 and 62 volatile compounds were detected, respectively. The most abundant group of flavour compounds at the end of the manufacturing process was hydrocarbons in the three feeding systems, followed by aldehydes, ketones and alcohols. Discriminant analysis selected six variables (dodecane, butadienol, pentenol, 2-pentenal, decen-3-ona and pyridine-2-methyl) and calculated two discriminating functions which allowed verification of chestnut in the finishing diet.     

Phenolic compounds and abscisic acid as potential markers for the floral origin of two Polish unifloral honeys

The phenolic profiles of Polish honey samples from heather (Calluna vulgaris L.) and buckwheat (Fagopyrum esculentum L.) were determined. The phenolic components were isolated from honey samples using Amberlite-XAD-2 as a solid-phase extraction sorbent. For the determination of the composition of the honey extracts HPLC with photodiode array detector was applied. Identification and quantification of phenolics was performed by comparison of their retention times and UV spectra with those of standard solutions of pure reference substances and by using those references as external standards. From among 20 commercially available standards over 15 of them were identified as present in all analysed samples. Taking into account the samples of the same unifloral honeys, similar qualitative but slightly quantitatively different phenolic characteristic profiles were observed. These profiles might be considered as “fingerprints” of heather and buckwheat honeys.     

添加大豆分离蛋白对羊乳干酪风味物质的影响

以新鲜羊乳为原料,直投式乳酸菌混合菌种为主发酵剂,费氏丙酸杆菌为次级发酵剂,添加不同比例(0、4%、10%m/m)大豆分离蛋白(soy protein isolate,SPI)制作混合型干酪。完全成熟之后利用固相微萃取(SPME)萃取富集干酪挥发性物质、气相色谱-质谱联用(GC/MS)检测干酪风味物质。以研究添加大豆分离蛋白对干酪风味物质的影响,确定适宜添加比例。实验结果表明:干酪中主要风味物质是酸类,其次是酮,醇,醛,酯类化合物;添加大豆分离蛋白使干酪中酸类、酮类化合物种类及含量明显减少;醇类化合物增加。大豆分离蛋白添加比为4%时,干酪有浓郁的奶香及黄油香气,羊乳膻味减少,并检出苯酚、吡咯等化合物,提高了干酪风味物质种类的丰富性。     

Analysis of volatile compounds of eucalyptus honey by solid phase extraction followed by gas chromatography coupled to mass spectrometry

Solid-phase extraction (SPE) followed by gas chromatography coupled to mass spectrometry has been used for the analysis of volatile compounds of eucalyptus honey. Different amounts of honey and dichloromethane were employed to optimize solid-phase extraction conditions. The best result was obtained with 20 g of honey eluted with 60 mL of dichloromethane. This method presented a good precision for volatile compounds usually found in honeys and allowed the quantification of 35 volatile compounds (terpenes and derivatives, furan and pyran compounds, ketones, benzene compounds, acids, and norisoprenoids) of eucalyptus honey.     

水煮鹅肉汤挥发性风味成分的GC/MS分析

采用共蒸馏分离和GC／MS分析，分析了水煮鹅肉汤的挥发性风味成分，结果共鉴定出50种成分，其中大部分为饱和或不饱和的醛酮类，其余主要为酯类和烷烃，另有少量的醇和酸。对比肉类挥发性风味成分的域值，对鹅肉汤风味贡献较大的成分主要为2，4-癸二烯醛、2-癸烯醛辛烯醛、壬醛、壬烯醛、壬二烯醛等。     

加工工艺对菜籽油主要挥发性风味成分的影响

采用固相微萃取（Solid Phase Microextraction,SPME）方法顶空萃取富集预榨菜籽毛油、浸出菜籽毛油、一级菜籽油、冷榨菜籽油和脱皮冷榨菜籽油中的挥发性成分,经气相色谱-质谱联用仪（Gas Chromatography-Mass Spectrometry,GC-MS）检测和初步分析发现,硫甙降解产物、氧化挥发物（醛、醇、烃、酮等）、杂环类物质是构成菜籽油的主要挥发性风味成分;2-甲代-1-丙烯基-氰、5-己腈、1-丁烯基-异硫氰酸酯、苯基丙氰、2-苯基乙基异硫氰酸酯是主要硫甙降解产物;反2-反4-庚二烯醛、反2-反4-癸二烯醛、4-羟基-3,5-二甲氧基苯甲醛、壬醛等醛类物质是氧化挥发物中的主要成分。加工工艺对菜籽油风味影响显著,经高温蒸炒、压榨获得的预榨毛油中杂环类物质种类和相对含量明显高于其它菜籽油。毛油经脱胶、脱酸、脱色、脱臭、脱水等处理后,一级油中产生的醛、醇、酮、烃等氧化挥发物种类增多,相对含量提高;在一级油中未检测到硫甙降解产物。     

Comparison of HS-SPME with hydrodistillation and SFE for the analysis of the volatile compounds of Zisu and Baisu,two varietal species of Perilla frutescens of Chinese origin

Content and composition of the volatiles of Baisu and Zisu, two varieties of Perilla frutescens, obtained by SFE, HS-SPME, and hydrodistillation, were compared. A total of 64 compounds were identified by GC/MS. Perillaldehyde (21.9–58.1%) and perilla ketone (54.5–83.5%) were the main volatile components of Zisu and Baisu, respectively. They might be partly responsible for the aroma and taste of perilla. The abundance of perilla ketone indicates that it is not safe to use Baisu as vegetable, and Baisu should not be used as substitute for Zisu. SFE extracted oil with high yield, and extracted some high molecular-weight compounds that did not contribute to the aroma. The HS-SPME was simple and sensitive for the analysis of volatile compounds of fresh perilla. The results showed that the drying process influenced the volatile components of perilla.     

不同产地香椿籽风味物质提取及成分分析

比较了从河南、湖北、陕西三地采集的香椿籽样品的挥发油含量和化学成分的差异。运用同步蒸馏萃取法提取香椿籽挥发油，所得挥发油通过GC／MS分析，并与Nist98标准图谱进行对照，定性分析了三种不同产地香椿籽挥发油，共鉴定出52种化学成分，三种挥发油均含有的主要成分是α波旁烯、γ-榄香烯、石竹烯及马兜铃烯。为香椿籽挥发油药用价值的进一步开发奠定了基础。