Glycerol Assay Kit

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00:09  Introduction
00:58   Principle
01:56    Reagent Preparation
02:39   Procedure
06:26   Calculations

Glycerol Assay Kit K-GCROL Scheme
Reference code: K-GCROL
SKU: 700004292

70 assays (manual) / 700 assays (microplate)

Content: 70 assays (manual) / 700 assays (microplate)
Shipping Temperature: Ambient
Storage Temperature: Short term stability: 2-8oC,
Long term stability: See individual component labels
Stability: > 2 years under recommended storage conditions
Analyte: Glycerol
Assay Format: Spectrophotometer, Microplate
Detection Method: Absorbance
Wavelength (nm): 340
Signal Response: Decrease
Linear Range: 0.8 to 35 µg of glycerol per assay
Limit of Detection: 0.34 mg/L
Reaction Time (min): ~ 5 min
Application examples: Wine (and grape juice), beer, spirits, vinegar, marzipan, fruit juices, soft drinks, toothpaste, honey, tobacco, paper (and cardboard), cosmetics, pharmaceuticals, soap and other materials (e.g. biological cultures, samples, etc.).
Method recognition: Methods based on this principle have been accepted by OIV and MEBAK

The Glycerol test kit is a simple, reliable, rapid and accurate method for the measurement and analysis of Glycerol in beverages, foodstuffs and other materials.

Note for Content: The number of manual tests per kit can be doubled if all volumes are halved.  This can be readily accommodated using the MegaQuantTM  Wave Spectrophotometer (D-MQWAVE).

Browse our wide range of alcohol assay kit products.

Scheme-K-GCROL GCROL Megazyme

  • Novel format for increased stability 
  • Very competitive price (cost per test) 
  • All reagents stable for > 2 years as supplied 
  • Very rapid reaction 
  • Mega-Calc™ software tool is available from our website for hassle-free raw data processing 
  • Standard included 
  • Suitable for manual and microplate formats
  • Extended cofactors stability
Certificate of Analysis
Safety Data Sheet
FAQs Assay Protocol Data Calculator Product Performance Validation Report
Megazyme publication

Megazyme “advanced” wine test kits general characteristics and validation.

Charnock, S. J., McCleary, B. V., Daverede, C. & Gallant, P. (2006). Reveue des Oenologues, 120, 1-5.

Many of the enzymatic test kits are official methods of prestigious organisations such as the Association of Official Analytical Chemicals (AOAC) and the American Association of Cereal Chemists (AACC) in response to the interest from oenologists. Megazyme decided to use its long history of enzymatic bio-analysis to make a significant contribution to the wine industry, by the development of a range of advanced enzymatic test kits. This task has now been successfully completed through the strategic and comprehensive process of identifying limitations of existing enzymatic bio-analysis test kits where they occurred, and then using advanced techniques, such as molecular biology (photo 1), to rapidly overcome them. Novel test kits have also been developed for analytes of emerging interest to the oenologist, such as yeast available nitrogen (YAN; see pages 2-3 of issue 117 article), or where previously enzymes were simply either not available, or were too expensive to employ, such as for D-mannitol analysis.

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Megazyme publication

Grape and wine analysis: Oenologists to exploit advanced test kits.

Charnock, S. C. & McCleary, B. V. (2005). Revue des Enology, 117, 1-5.

It is without doubt that testing plays a pivotal role throughout the whole of the vinification process. To produce the best possible quality wine and to minimise process problems such as “stuck” fermentation or troublesome infections, it is now recognised that if possible testing should begin prior to harvesting of the grapes and continue through to bottling. Traditional methods of wine analysis are often expensive, time consuming, require either elaborate equipment or specialist expertise and frequently lack accuracy. However, enzymatic bio-analysis enables the accurate measurement of the vast majority of analytes of interest to the wine maker, using just one piece of apparatus, the spectrophotometer (see previous issue No. 116 for a detailed technical review). Grape juice and wine are amenable to enzymatic testing as being liquids they are homogenous, easy to manipulate, and can generally be analysed without any sample preparation.

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Biocontrol using Torulaspora delbrueckii in Sequential Fermentation: New Insights into Low Sulfites Verdicchio Wines.

Canonico, L., Agarbati, A., Galli, E., Comitini, F. & Ciani, M. (2023). Foods, 12(15), 2899.

Torulaspora delbrueckii showed renewed interest in recent years in the fermentation of wine, for its biotechnological potential linked to the ability to enhance flavor and aroma and it probably is the non-Saccharomyces yeast currently widely used in winemaking. On the base of this, sequential fermentations with a selected native strain of T. delbrueckii (DiSVA 130) and low sulfite native strain of Saccharomyces cerevisiae (DiSVA 709) were carried out to establish their contribution in biocontrol and aroma profile. A first set trials, carried out in winery, were set up to establish the effect of the sulfur dioxide addition on pure and T. debrueckii/S. cerevisiae sequential fermentations. A second set of sequential fermentations without SO2 addition were conducted in the same conditions, to evaluate the biocontrol and aromatic effectiveness of the T. delbrueckii native strain and a commercial one. The effective biocontrol action of native T. delbrueckii inoculated in sequential fermentation was shown, indeed without SO2 addition the presence of native T. delbrueckii revealed an effective fungistatic action in the first two days of fermentation. Moreover, the native T. delbrueckii strain seems to have fermentative performances comparable to those of T. delbrueckii commercial strain showing a more evident biocontrol action (wild yeasts reduced by c.a. 1 Log at 2nd day) and its presence did not negatively affect S. cerevisiae fermentation activity. Finally, the combination of both native and commercial T. delbrueckii/S. cerevisiae trials led distinctive aromatic profile of wines with a significant enhancement of isoamyl acetate, phenyl ethyl acetate, supported by positive appreciations, from the tasters, for ripe and tropical fruits, citrus and balance. The whole results indicate that the proposed strain could be a potential biocontrol tool toward wild yeasts in the first phase of fermentation also contributing to improve and differentiate the final aroma wine.

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Economical Di-Rhamnolipids Biosynthesis by Non-Pathogenic Burkholderia thailandensis E264 Using Post-Consumption Food Waste in a Biorefinery Approach.

Kumar, R., Johnravindar, D., Wong, J. W., Patria, R. D. & Kaur, G. (2023). Sustainability, 15(1), 59.

Rhamnolipids (RLs) are one of the most promising eco-friendly green alternatives to commercially viable fossil fuel-based surfactants. However, the current bioprocess practices cannot meet the required affordability, quantity, and biocompatibility within an industrially relevant framework. To circumvent these issues, our study aims to develop a sustainable biorefinery approach using post-consumption food waste as a second-generation feedstock. In-depth substrate screening revealed that food waste hydrolysate (FWH) was rich in readily assimilable carbohydrates, volatile fatty acids, and amino acids. The fermentative valorization of FWH as a sole carbon and energy source with Burkholderis thailandensis E264 in a bioreactor showed active RLs biosynthesis of up to 0.6–0.8 g/L (34–40 mg/g FWH) in a short duration (72 h). In terms of the kinetic parameters, the FWH-RLs outperformed other supplemented pure/waste streams. Interestingly, the recovered RLs had a long chain length, with Rha-Rha-C12-C14 being the predominant isoform and exhibiting a strong emulsification ability (E24, 54.6%). To the best of our knowledge, this study is the first to prove bioreactor-level RLs production and their abundance in food waste. Moreover, the feasibility of this developed process could propel next-generation biosurfactants, lower waste burdens, and increase the industrial applicability of RLs, thereby significantly contributing to the development of a circular bioeconomy.

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Saccharomyces uvarum yeast isolate consumes acetic acid during fermentation of high sugar juice and juice with high starting volatile acidity.

Inglis, D., Kelly, J., van Dyk, S., Dowling, L., Pickering, G. & Kemp, B. (2020). OENO One, 54(2).

Aim: A Saccharomyces uvarum isolate was assessed for its ability to metabolize acetic acid present in juice and during the fermentation of partially dehydrated grapes. The impact on other yeast metabolites was also compared using an S. uvarum isolate and an S. cerevisiae wine yeast. The upper limit of fruit concentration that allowed the S. uvarum isolate to ferment wines to < 5 g/L residual sugar was defined. Methods and results: Cabernet franc grapes were partially dehydrated to three different post-harvest sugar targets (24.5 °Brix, 26.0 °Brix, and 27.5 °Brix) along with non-dehydrated grapes (21.5 °Brix control). Musts from all treatments were vinified with either the S. uvarum isolate CN1, formerly identified as S. bayanus, or S. cerevisiae EC1118. All wines were successfully vinified to less than 5 g/L residual sugar. Fermentation kinetics between the two yeasts were similar for all wines other than 27.5 °Brix, where CN1 took three days longer. During fermentation with CN1, acetic acid peaked on day two, then decreased in concentration, resulting in final wine acetic acid lower than that measured on day two. Wines fermented with EC1118 showed an increase in acetic acid over the time-course of fermentation. Significantly lower wine oxidative compounds (acetic acid, acetaldehyde and ethyl acetate) and higher glycerol resulted in wine produced with CN1 in comparison to EC1118. Both yeasts produced comparable ethanol at each Brix level tested. Further studies showed that CN1 lowered acetic acid seven-fold from 0.48 g/L in juice to 0.07 g/L in wine whereas EC1118 reduced acetic acid to 0.18 g/L. Conclusions: The autochthonous S. uvarum yeast isolate successfully fermented partially dehydrated grapes to < 5 g/L sugar up to 27.5 ºBrix. The consumption rate of acetic acid was faster than its production during fermentation, resulting in low acetic acid, acetaldehyde and ethyl acetate in wine in comparison to a commercial S. cerevisiae yeast while consistently producing higher glycerol. Significance and impact of the study: The S. uvarum yeast isolate can metabolize acetic acid during fermentation to significantly lower acetic acid, ethyl acetate and acetaldehyde in wine. It can also reduce acetic acid by seven-fold from the starting juice to the finished wine, which could have potential application for managing sour rot arising in the vineyard or during the dehydration process in making appassimento-style wines.

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Selection of low nitrogen demand yeast strains and their impact on the physicochemical and volatile composition of mead.

Schwarz, L. V., Marcon, A. R., Delamare, A. P. L., Agostini, F., Moura, S. & Echeverrigaray, S. (2020). Journal of Food Science and Technology, 1-12.

Mead is an ancient alcoholic beverage produced through the fermentation of a diluted solution of honey. Due to the peculiar and varied composition of honey, mead production faces several problems, such as slow or stuck fermentations mainly due to the low nitrogen concentration, lack of uniformity of the final product and the production of unpleasant aromas. In this context, this work aimed to select low nitrogen-demand yeast strains and evaluate their potential for the production of mead. Therefore, among 21 commercial wine yeast strains, 5 were selected based on their fermentative behavior at low assimilable nitrogen concentrations. The selected strains were further evaluated for their contributions in meads produced with limited nitrogen availability, and the results showed significant differences on some physicochemical parameters like biomass production, residual sugars, glycerol concentration, and fermentative rate. Moreover, meads obtained with selected strains differed in the concentration of several volatile compounds. The volatile compounds concentration and the principal component analysis based on odor activity values allowed separating strains into three groups. In general, S. cerevisiae var bayanus strains (QA23, Spark, and AWRI-R2) were the largest producers of aromatic compounds, particularly those with floral and fruity descriptors. The selection of yeast strains with low nitrogen-demand and different volatile compounds production can be explored by mead makers to limit fermentation problems and obtain characteristic products.

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The Use of CRISPR-Cas9 Genome Editing to Determine the Importance of Glycerol Uptake in Wine Yeast During Icewine Fermentation.

Muysson, J., Miller, L., Allie, R. & Inglis, D. L. (2019). Fermentation, 5(4), 93.

The high concentration of sugars in Icewine juice causes formidable stress for the fermenting Saccharomyces cerevisiae, causing cells to lose water and shrink in size. Yeast can combat this stress by increasing the internal concentration of glycerol by activating the high osmolarity glycerol response to synthesize glycerol and by actively transporting glycerol into the cell from the environment. The H+/glycerol symporter, Stl1p, has been previously characterized as being glucose repressed and inactivated, despite osmotic stress induction. To further investigate the role of Stl1p in Icewine fermentations, we developed a rapid single plasmid CRISPR-Cas9-based genome editing method to construct a strain of the common Icewine yeast, S. cerevisiae K1-V1116, that lacks STL1. In an Icewine fermentation, the ∆STL1 strain had reduced fermentation performance, and elevated glycerol and acetic acid production compared to the parent. These results demonstrate that glycerol uptake by Stl1p has a significant role during osmotically challenging Icewine fermentations in K1-V1116 despite potential glucose downregulation.

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Metschnikowia pulcherrima selected strain for ethanol reduction in wine: Influence of cell immobilization and aeration condition.

Canonico, L., Comitini, F. & Ciani, M. (2019). Foods, 8(9), 378.

One of the most important problems in the winemaking field is the increase of ethanol content in wine. Wines with high ethanol level negatively affect wine flavor and human health. In this study, we evaluated the use of a selected strain of Metschnikowia pulcherrima in immobilized form and under different aeration conditions, to reduce the ethanol content evaluating the volatile profile of the resulting wines. In a preliminary screening the best conditions regarding free/immobilized cells, static/aerated fermentation and inoculation level were identified. Bench-Top fermentation trials with different aeration conditions showed that the use of M. pulcherrima selected strain with aeration flow of 20 mL/L/min during the first 72 h of fermentation, led an ethanol reduction of 1.38% (v/v) in comparison with Saccharomyces cerevisiae control strain. The analytical profile of the resulting wines did not show any negative feature. Indeed, the concentration of ethyl acetate, that above its sensory threshold impacts negatively the wine sensory profile, was found at an acceptable level. On the other hand, an increase in the concentration of significant fruity and flower compounds was found.

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Specific molecular interactions between vitis vinifera and botrytis cinerea are required for noble rot development in grape berries.

Lovato, A., Zenoni, S., Tornielli, G. B., Colombo, T., Vandelle, E. & Polverari, A. (2019). Postharvest Biology and Technology, 156, 110924.

Under peculiar climatic conditions, the beneficial form of the necrotrophic fungus Botrytis cinerea can develop on grape berries as a latent infection, known as noble rot, which induces positive biochemical and metabolic changes in the berries, including an increase in the sugar content and the production of aromatic compounds that improve wine quality. The infected berries undergo rapid withering, which is required to produce famous sweet white wines such as Sauternes and Tokaj. To gain insight into the molecular interactions between grapevine berries (Vitis vinifera) and B. cinerea during the establishment of noble rot, we prepared a large-scale transcriptomics dataset representing noble rot development and carried out a comparative meta-analysis with gray mold infection and natural post-harvest withering. In particular, we artificially induced berry botrytization of two grape varieties (Garganega and Möller-Thurgau) and sampled them at different stages of noble rot for comparative whole-transcriptome analysis, highlighting important common transcriptional reprogramming in both varieties reflecting an accelerated withering process. Simultaneously, we analyzed the modulation of B. cinerea genes and compared the expression profile during noble rot development with the previously reported gray mold infection profile, revealing the onset of an infection process by the fungus in its beneficial form associated with reduced virulence. This, together with the restrained plant defense response observed in botrytized berries, may favour the development of noble rot instead of gray mold. Finally, the comprehensive meta-analysis of gene expression during noble rot infection, gray mold and post-harvest withering led to the identification of key genes specifically modulated during noble rot infection.

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Coordinated Transcriptional Control of Adipocyte Triglyceride Lipase (Atgl) by transcription factors Sp1 and PPARγ during Adipocyte Differentiation.

Roy, D., Farabaugh, K. T., Wu, J., Charrier, A., Smas, C., Hatzoglou, M., Thirumurgan, K. & Buchner, D. A. (2017). Journal of Biological Chemistry, jbc-M117.

The breakdown of stored fat deposits into its components is a highly regulated process that maintains plasma levels of free fatty acids to supply energy to cells. Insulin-mediated transcription of Atgl, the enzyme that mediates the rate-limiting step in lipolysis, is a key point of this regulation. In conditions such as obesity or insulin resistance, Atgl transcription is often misregulated, which can contribute to overall disease progression. The mechanisms by which Atgl is induced during adipogenesis are not fully understood. We utilized computational approaches to identify putative transcriptional regulatory elements in Atgl and then tested the effect of these elements and the transcription factors that bind to them in cultured pre- and mature adipocytes. Herein, we report that Atgl is downregulated by the basal transcription factor Sp1 in preadipocytes, and that the magnitude of downregulation dependents on interactions between Sp1 and PPARγ. In mature adipocytes, when PPARγ is abundant, PPARγ abrogated the transcriptional repression by Sp1 at the Atgl promoter and upregulated Atgl mRNA expression. Targeting the PPARγ-Sp1 interaction could be a potential therapeutic strategy to restore insulin sensitivity by modulating Atgl levels in adipocytes.

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Cytosolic Redox Status of Wine Yeast (Saccharomyces Cerevisiae) under Hyperosmotic Stress during Icewine Fermentation.

Yang, F., Heit, C. & Inglis, D. L. (2017). Fermentation, 3(4), 61.

Acetic acid is undesired in Icewine. It is unclear whether its production by fermenting yeast is linked to the nicotinamide adenine dinucleotide (NAD+/NADH) system or the nicotinamide adenine dinucleotide phosphate (NADP+/NADPH) system. To answer this question, the redox status of yeast cytosolic NAD(H) and NADP(H) were analyzed along with yeast metabolites to determine how redox status differs under Icewine versus table wine fermentation. Icewine juice and dilute Icewine juice were inoculated with commercial wine yeast Saccharomyces cerevisiae K1-V1116. Acetic acid was 14.3-fold higher in Icewine fermentation than the dilute juice condition. The ratio of NAD+ to total NAD(H) was 24-fold higher in cells in Icewine fermentation than the ratio from the dilute juice condition. Conversely, the ratio of NADP+ to total NADP(H) from the dilute fermentation was 2.9-fold higher than that in the Icewine condition. These results support the hypothesis that in Icewine, increased NAD+ triggered the catalysis of NAD+-dependent aldehyde dehydrogenase(s) (Aldp(s)), which led to the elevated level of acetic acid in Icewine, whereas, in the dilute condition, NADP+ triggered NADP+-dependent Aldp(s), resulting in a lower level of acetic acid. This work, for the first time, analyzed the yeast cytosolic redox status and its correlation to acetic acid production, providing a more comprehensive understanding of the mechanism of acetic acid production in Icewine.

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Chemical composition and in vitro antimicrobial and cytotoxic activities of plum (Prunus domestica L.) wine.

Miljić, U., Puškaš, V., Velićanski, A., Mašković, P., Cvetković, D. & Vujić, J. (2016). Journal of the Institute of Brewing, 122(2), 342-349.

A moderate intake of wine is associated with a positive impact on human health owing to the effects of important biologically active components present in the wine in large amounts. The aim of this study was to examine the chemical composition and to assess antimicrobial and cytotoxic activities of fruit wines produced from three plum varieties (Čačanska rana, Čačanska lepotica and Požegača) commonly grown in Serbia as an approach to assess the quality and acceptability of these wines as a functional food. Furthermore, the activity of a series of control samples was assessed in order to determine components from the wine that are responsible for its functional properties. The plum wines produced showed considerable antimicrobial activity against six bacterial and two yeast strains used in this study. In addition to antimicrobial activity, the plum wines showed a significant cytotoxic effect (IC50 < 50 µg Ml-1) on the growth of three tested cancer cell lines (Hep2c, RD and L2OB). Regarding the determined activities, Čačanska rana plum wine achieved the best results. The results indicated that the antimicrobial activity of the plum wines was, in large part, based on the effects of the total acids and the pH value, while the contribution of ethanol and the content of the phenolic compounds were not significant. Similar conclusions were drawn regarding the cytotoxic activity of this fruit wine. The results can be seen as a contribution to the global acceptance of fruit wines as a functional food, with the accent placed on moderate consumption. An important advantage of fruit wines (in particular plum wine), compared with traditional grape wine, is their lower alcohol content.

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Isolation and characterisation of a heat-resistant peptidase from Pseudomonas panacis withstanding general UHT processes.

Baur, C., Krewinkel, M., Kutzli, I., Kranz, B., von Neubeck, M., Huptas, C., Wenning, M., Scherer, S., Stoeckel, M., Hinrichs, J., Stressler, T. & Stressler, T. (2015). International Dairy Journal, 49, 46-55.

A secreted peptidase from Pseudomonas panacis was identified and purified. Genome sequencing of the producer strain allowed identification of the peptidase as AprA based on a comparison to peptide sequences of mass spectra obtained from the purified enzyme. The amino acid sequence of the 49.4 kDa peptidase was 98% similar to the metallopeptidase AprX from a Pseudomonas fluorescens strain. The peptidase showed maximum activity at pH 8 and 40°C and withstood general ultra-high temperature (UHT) processing (138°C for 18 s) in skim milk, with 88.0 ± 7.7% of the initial enzyme activity remaining after heating. The peptidase showed considerable enzyme activity under storage conditions of UHT milk. The potential for spoilage of milk might during storage was verified by adding very low enzyme activities to UHT-treated milk. The addition of 1 pkat mL-1 peptidase activity resulted in a destabilisation of the milk during four weeks storage.

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Rapid Assessment of Gray Mold (Botrytis cinerea) Infection in Grapes Using Biosensors System.

Cinquanta, L., Albanese, D., De Curtis, F., Malvano, F., Crescitelli, A. & Di Matteo, M. (2015). American Journal of Enology and Viticulture, ajev-2015.

Botrytis cinerea is responsible for the gray mold disease, which causes considerable economic losses for winemakers. Its evaluation in wine grapes is commonly performed through visual estimation, which was demonstrated to be prone to assessor bias. Rapid and simple enzymatic carbon screen printed amperometric biosensors were here used to evaluate gluconic acid and glycerol content on wine grapes at different B. cinerea infection degrees. The lower concentrations measurable by screen-printed amperometric biosensors were 3 mg/L for gluconic acid (corresponding to an infection degree lower than 1%) and 35 mg/L for glycerol; the response times with a flow rate of 0.5 mL/min were in a range of 0.5 to 2 min in the linear ranges. This study demonstrates the effectiveness of the biosensors for rapid analysis of gluconic acid and glycerol in grapes, confirming their high correlation with B. cinerea degree of infection (R2 = 0.98). Thus, the biosensor developed to measure gluconic acid in grapes (or must), was more precise, and gave a faster response than methods that currently exist allowing the percentage of infection of grape berries by B. cinerea to be evaluated.

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Aroma compounds in Ontario Vidal and Riesling icewines. I. Effects of harvest date.

Bowen, A. J. & Reynolds, A. G. (2015). Food Research International, 76, 540-549.

Icewine is a sweet dessert wine made from pressing grapes naturally frozen on the vines. It is likely that freeze/thaw cycles endured by icewine grapes change their chemical and sensory profiles due to climatic events. Our objective was to determine the influence of harvest date on icewine must and wine basic chemical variables and aroma compounds. Riesling and Vidal icewines were made from grapes picked between December 2004 and February 2005; Harvest 1 (H1): 19 December; Harvest 2: 29 December; Harvest 3 (H3): 18 January; and Harvest 4 (H4): 11 February (Vidal only). Icewine musts differed in titratable acidity and pH (Vidal only). All basic wine chemical analytes differed across harvest dates. All aroma compounds differed in Vidal and Riesling wines. Highest concentrations for most aroma compounds were in the last harvest date; 16 of 24 for Vidal and 17 of 23 for Riesling. The latest harvest date had highest ethyl isobutyrate, ethyl 3-methylbutyrate, 1-hexanol, 1-octen-3-ol, 1-octanol, cis-rose oxide, nerol oxide, ethyl benzoate, ethyl phenylacetate, γ-nonalactone and β-damascenone. H1 had highest ethyl butyrate, ethyl hexanoate, linalool, 4-vinylguaiacol and ethyl octanoate. Based on odor activity values, the most odor-potent compounds were β-damascenone, cis-rose oxide, 1-octen-3-ol, ethyl octanoate, ethyl hexanoate, and 4-vinylguaiacol across harvest dates. PCA found most aroma compounds associated with the last harvest date, 4-vinylguaicol excepted, which was associated with H1. Harvest date was considered a discriminating dimension using canonical variant analysis for volatile compounds.

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Safety Information
Symbol : GHS08
Signal Word : Danger
Hazard Statements : H334
Precautionary Statements : P261, P284, P304+P340, P342+P311, P501
Safety Data Sheet
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