Name: profiling volatile compounds in blackcurrant fruit using headspace solid-phase microextraction coupled to gas chromatography-mass spectrometry
Uploaded: 2021-06-09T14:00:00
Description: Watch this Scientific Journal Video about Profiling Volatile Compounds in Blackcurrant Fruit using Headspace Solid-Phase Microextraction Coupled to Gas Chromatography-Mass Spectrometry at JoVE.com

The authors thank the Servicios Centrales de Apoyo a la Investigaci&#243;n from University of Malaga for HS-SPME/GC-MS measurements. We acknowledge the assistance of Sara Fern&#225;ndez-Palacios Campos in volatile quantification. We also thanks GoodBerry&#180;s consortium members for providing the fruit material.

1. Fruit harvesting
<ol>
	<li>Grow between 4 to 6 plants per genotype and/or treatment to ensure sufficient fruit material and variability.</li>
	<li>If possible, harvest the samples on the same date; if there is not enough fruit material, pool together samples harvested on different dates. 
	NOTE: It is recommended that the harvest time (morning, noon, afternoon) remains approximately identical as VOC profiles are affected by daytime/circadian rhythm28,29...

<ol>
	<li>Klee, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improving+the+flavor+of+fresh+fruits:+Genomics,+biochemistry,+and+biotechnology.">Improving the flavor of fresh fruits: Genomics, biochemistry, and biotechnology.</a> New Phytologist. 187 (1), 44-56 (2010).</li><li>Ferr&#227;o, L. F. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genome-wide+association+of+volatiles+reveals+candidate+loci+for+blueberry+flavor.">Genome-wide association of volatiles reveals candidate loci for blueberry flavor.</a> New Phytologist. 226 (6), 1725-1737 (2020).</li><li>Klee, H. J., Tieman, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genetics+of+fruit+flavour+preferences.">The genetics of fruit flavour preferences.</a> Nature Reviews Genetics. 19, 347-356 (2018).</li><li>Vallarino, J. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+quantitative+trait+loci+and+candidate+genes+for+primary+metabolite+content+in+strawberry+fruit.">Identification of quantitative trait loci and candidate genes for primary metabolite content in strawberry fruit.</a> Horticulture Research. 6, 4 (2019).</li><li>Jung, K., Fastowski, O., Poplacean, I., Engel, K. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+and+sensory+evaluation+of+volatile+constituents+of+fresh+blackcurrant+(Ribes+nigrum+L.)+fruits.">Analysis and sensory evaluation of volatile constituents of fresh blackcurrant (Ribes nigrum L.) fruits.</a> Journal of Agricultural and Food Chemistry. 65 (43), 9475-9487 (2017).</li><li>Vallarino, J. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+diversity+of+strawberry+germplasm+using+metabolomic+biomarkers.">Genetic diversity of strawberry germplasm using metabolomic biomarkers.</a> Scientific Reports. 8, 14386 (2018).</li><li>Zhang, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insights+into+the+major+aroma-active+compounds+in+clear+red+raspberry+juice+(Rubus+idaeus+L.+cv.+Heritage)+by+molecular+sensory+science+approaches.">Insights into the major aroma-active compounds in clear red raspberry juice (Rubus idaeus L. cv. Heritage) by molecular sensory science approaches.</a> Food Chemistry. 336, 127721 (2021).</li><li>Farneti, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploring+blueberry+aroma+complexity+by+chromatographic+and+direct-injection+spectrometric+techniques.">Exploring blueberry aroma complexity by chromatographic and direct-injection spectrometric techniques.</a> Frontiers in Plant Science. 8, 617 (2017).</li><li>Tikunov, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genetic+and+functional+analysis+of+flavor+in+commercial+tomato:+the+FLORAL4+gene+underlies+a+QTL+for+floral+aroma+volatiles+in+tomato+fruit.">The genetic and functional analysis of flavor in commercial tomato: the FLORAL4 gene underlies a QTL for floral aroma volatiles in tomato fruit.</a> The Plant Journal. 103 (3), 1189-1204 (2020).</li><li>Tieman, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+chemical+genetic+roadmap+to+improved+tomato+flavor.">A chemical genetic roadmap to improved tomato flavor.</a> Science. 355 (6323), 391-394 (2017).</li><li>S&#225;nchez-Sevilla, J. F., Cruz-Rus, E., Valpuesta, V., Botella, M. A., Amaya, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deciphering+gamma-decalactone+biosynthesis+in+strawberry+fruit+using+a+combination+of+genetic+mapping,+RNA-Seq+and+eQTL+analyses.">Deciphering gamma-decalactone biosynthesis in strawberry fruit using a combination of genetic mapping, RNA-Seq and eQTL analyses.</a> BMC Genomics. 15, 218 (2014).</li><li>Kumar, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genome-wide+scans+reveal+genetic+architecture+of+apple+flavour+volatiles.">Genome-wide scans reveal genetic architecture of apple flavour volatiles.</a> Molecular Breeding. 35, 118 (2015).</li><li>Bauchet, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+major+loci+and+genomic+regions+controlling+acid+and+volatile+content+in+tomato+fruit:+implications+for+flavor+improvement.">Identification of major loci and genomic regions controlling acid and volatile content in tomato fruit: implications for flavor improvement.</a> New Phytologist. 215 (2), 624-641 (2017).</li><li>S&#225;nchez, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+integrative+'+omics'+approach+identifies+new+candidate+genes+to+impact+aroma+volatiles+in+peach+fruit.">An integrative ' omics' approach identifies new candidate genes to impact aroma volatiles in peach fruit.</a> BMC Genomics. 14, 343 (2013).</li><li>Hummer, K. E., Dale, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Horticulture+of+Ribes.">Horticulture of Ribes.</a> Forest Pathology. 40 (3-4), 251-263 (2010).</li><li>Vagiri, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenols+and+ascorbic+acid+in+black+currants+(Ribes+nigrum+L.):+Variation+due+to+genotype,+location,+and+year.">Phenols and ascorbic acid in black currants (Ribes nigrum L.): Variation due to genotype, location, and year.</a> Journal of Agricultural and Food Chemistry. 61 (39), 9298-9306 (2013).</li><li>Marsol-Vall, A., Kortesniemi, M., Karhu, S. T., Kallio, H., Yang, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Profiles+of+volatile+compounds+in+blackcurrant+(Ribes+nigrum)+cultivars+with+a+special+focus+on+the+influence+of+growth+latitude+and+weather+conditions.">Profiles of volatile compounds in blackcurrant (Ribes nigrum) cultivars with a special focus on the influence of growth latitude and weather conditions.</a> Journal of Agricultural and Food Chemistry. 66 (28), 7485-7495 (2018).</li><li>Varming, C., Petersen, M. A., Poll, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+isolation+methods+for+the+determination+of+important+aroma+compounds+in+black+currant+(Ribes+nigrum+L.)+juice,+using+nasal+impact+frequency+profiling.">Comparison of isolation methods for the determination of important aroma compounds in black currant (Ribes nigrum L.) juice, using nasal impact frequency profiling.</a> Journal of Agricultural and Food Chemistry. 52 (6), 1647-1652 (2004).</li><li>Andersson, J., von Sydow, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+aroma+of+black+currants+I.+Higher+boiling+compounds.">The aroma of black currants I. Higher boiling compounds.</a> Acta Chemica Scandinavica. 18, 1105-1114 (1964).</li><li>Andersson, J., von Sydow, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+aroma+of+black+currants.+III.+Chemical+characterization+of+different+varieties+and+stages+of+ripeness+by+gas+chromatography.">The aroma of black currants. III. Chemical characterization of different varieties and stages of ripeness by gas chromatography.</a> Acta Chemica Scandinavica. 20, 529-535 (1966).</li><li>Andersson, J., von Sydow, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+aroma+of+black+currants+II.+Lower+boiling+compounds.">The aroma of black currants II. Lower boiling compounds.</a> Acta Chemica Scandinavica. 20, 522-528 (1966).</li><li>Marsol-Vall, A., Laaksonen, O., Yang, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+processing+and+storage+conditions+on+volatile+composition+and+odor+characteristics+of+blackcurrant+(Ribes+nigrum)+juices.">Effects of processing and storage conditions on volatile composition and odor characteristics of blackcurrant (Ribes nigrum) juices.</a> Food Chemistry. 293, 151-160 (2019).</li><li>Del Castillo, M. L. R., Dobson, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Varietal+differences+in+terpene+composition+of+blackcurrant+(Ribes+nigrum+L)+berries+by+solid+phase+microextraction/gas+chromatography.">Varietal differences in terpene composition of blackcurrant (Ribes nigrum L) berries by solid phase microextraction/gas chromatography.</a> Journal of the Science of Food and Agriculture. 82 (13), 1510-1515 (2002).</li><li>Azzi-Achkouty, S., Estephan, N., Ouaini, N., Rutledge, D. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Headspace+solid-phase+microextraction+for+wine+volatile+analysis.">Headspace solid-phase microextraction for wine volatile analysis.</a> Critical Reviews in Food Science and Nutrition. 57 (10), 2009-2020 (2017).</li><li>Vallarino, J. G., Antonio, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acquisition+of+volatiles+compounds+by+gas+chromatography-mass+spectrometry+(GC-MS).">Acquisition of volatiles compounds by gas chromatography-mass spectrometry (GC-MS).</a> Plant Metabolomics: Methods and Protocols. 1778, 225-239 (2018).</li><li>Moreira, N., Lopes, P., Cabral, M., Guedes de Pinho, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HS-SPME/GC-MS+methodologies+for+the+analysis+of+volatile+compounds+in+cork+material.">HS-SPME/GC-MS methodologies for the analysis of volatile compounds in cork material.</a> European Food Research and Technology. 242, 457-466 (2016).</li><li>Rambla, J. L., L&#243;pez-Gresa, M. P., Bell&#233;s, J. M., Granell, A., Alonso, J. M., Stepanova, A. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolomic+profiling+of+plant+tissues.">Metabolomic profiling of plant tissues.</a> Plant Functional Genomics and Protocols, Methods in Molecular Biology. 1284, 221-235 (2015).</li><li>Abbas, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volatile+terpenoids:+multiple+functions,+biosynthesis,+modulation+and+manipulation+by+genetic+engineering.">Volatile terpenoids: multiple functions, biosynthesis, modulation and manipulation by genetic engineering.</a> Planta. 246 (5), 803-816 (2017).</li><li>Kolosova, N., Gorenstein, N., Kish, C. M., Dudareva, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+circadian+methyl+benzoate+emission+in+diurnally+and+nocturnally+emitting+plants.">Regulation of circadian methyl benzoate emission in diurnally and nocturnally emitting plants.</a> Plant Cell. 13 (10), 2333-2347 (2001).</li><li>Dudareva, N., Pichersky, E., Gershenzon, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biochemistry+of+plant+volatiles.">Biochemistry of plant volatiles.</a> Plant Physiology. 135 (4), 1893-1902 (2004).</li><li>Borges, R. M., Ranganathan, Y., Krishnan, A., Ghara, M., Pramanik, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=When+should+fig+fruit+produce+volatiles?+Pattern+in+a+ripening+process.">When should fig fruit produce volatiles? Pattern in a ripening process.</a> Acta Oecologica. 37 (6), 611-618 (2011).</li><li>Jarret, D. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+transcript+and+metabolite+atlas+of+blackcurrant+fruit+development+highlights+hormonal+regulation+and+reveals+the+role+of+key+transcription+factors.">A transcript and metabolite atlas of blackcurrant fruit development highlights hormonal regulation and reveals the role of key transcription factors.</a> Frontiers in Plant Science. 9, 1-22 (2018).</li><li>Li, B., Lecourt, J., Bishop, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+non-destructive+early+assessment+of+fruit+ripeness+towards+defining+optimal+time+of+harvest+and+yield+prediction-a+review.">Advances in non-destructive early assessment of fruit ripeness towards defining optimal time of harvest and yield prediction-a review.</a> Plants. 7 (1), 3 (2018).</li><li>Ul-Hassan, M. N., Zainal, Z., Ismail, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Green+leaf+volatiles:+Biosynthesis,+biological+functions+and+their+applications+in+biotechnology.">Green leaf volatiles: Biosynthesis, biological functions and their applications in biotechnology.</a> Plant Biotechnology Journal. 13 (6), 727-739 (2015).</li><li>Gaston, A., Osorio, S., Denoyes, B., Rothan, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applying+the+Solanaceae+strategies+to+strawberry+crop+improvement.">Applying the Solanaceae strategies to strawberry crop improvement.</a> Trends in Plant Science. 25 (2), 130-140 (2020).</li><li>Gilbert, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identifying+breeding+priorities+for+blueberry+flavor+using+biochemical,+sensory,+and+genotype+by+environment+analyses.">Identifying breeding priorities for blueberry flavor using biochemical, sensory, and genotype by environment analyses.</a> PLoS ONE. 10 (9), 0138494 (2015).</li><li>Bueno, M., Resconi, V. C., Campo, M. M., Ferreira, V., Escudero, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+robust+HS-SPME-GC-MS+method+for+the+analysis+of+solid+food+samples.+Analysis+of+volatile+compounds+in+fresh+raw+beef+of+differing+lipid+oxidation+degrees.">Development of a robust HS-SPME-GC-MS method for the analysis of solid food samples. Analysis of volatile compounds in fresh raw beef of differing lipid oxidation degrees.</a> Food Chemistry. 281, 49-56 (2019).</li><li>Burzynski-Chang, E. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HS-SPME-GC-MS+analyses+of+volatiles+in+plant+populations-quantitating+compound+&#215;+individual+matrix+effects.">HS-SPME-GC-MS analyses of volatiles in plant populations-quantitating compound &#215; individual matrix effects.</a> Molecules. 23 (10), 2436 (2018).</li></ol>

Breeding for fruit aroma has long been hindered by the complex genetics and biochemistry underlying the synthesis of volatile compounds and the lack of technologies for proper phenotyping. However, recent advances in metabolomic platforms, combined with genomic tools, are finally allowing the identification of the metabolites responsible for consumer preferences and to breed crops with improved flavor3. While most progress has been achieved in the model fruit, tomato9,...

Flavor is an essential quality trait for any fruit, impacting consumer acceptance and thus significantly affecting marketability. Flavor perception involves a combination of the taste and olfactory systems and depends chemically on the presence and concentration of a wide range of compounds that accumulate in edible plant parts, or in case of VOCs, are emitted by the ripe fruit1,2. While traditional breeding has focused on agronomic traits such as yield and pest resistance, fruit quality trait improvement, including flavor, has long been neglected due to the genetic complexity and the difficulty to properly phenotype these characteristics, leading to consumer discontent3,4. Recent advances in metabolomic platforms have been successful in identifying and quantifying key compounds responsible for fruit taste and aroma5,6,7,8. Furthermore, the combination of metabolite profiling with genomic or transcriptomic tools allows the elucidation of the genetics underlying fruit flavor, which in turn will help breeding programs develop new varieties with enhanced organoleptic characteristics2,4,9,10,11,12,13,14.
Blackcurrant (Ribes nigrum) berries are highly appreciated for their flavor and nutritional properties, being widely cultivated across the temperate zones of Europe, Asia, and New Zealand15. Most of the production is processed for food products and beverages, which are very popular in the Nordic countries, mainly due to the berries&#39; organoleptic properties. The intense color and flavor of the fruit are the result of a combination of anthocyanins, sugars, acids, and VOCs present in the ripe fruits16,17,18. The analysis of blackcurrant volatiles goes back to the 1960s19,20,21. More recently, several studies have focused on blackcurrant VOCs, identifying important compounds for fruit aroma perception and assessing the impact of genotype, environment, or storage and processing conditions on VOC content5,17,18,22,23.
Because of its numerous advantages, the technique of choice for high-throughput volatile profiling is HS-SPME/GC-MS24,25. A silica fiber, coated with a polymeric phase, is mounted on a syringe device, allowing the adsorption of the volatiles in the fiber until an equilibrium phase is reached. Headspace extraction protects the fiber from the nonvolatile compounds present in the matrix24. SPME can successfully isolate a high number of VOCs present at highly variable concentrations (parts per billion to parts per million)25. In addition, it is a solvent-free technique that requires limited sample processing. Other advantages of HS-SPME are the ease of automation and its relatively low cost.
However, its success can be limited, depending on the chemical nature of the VOCs, the extraction protocol (including time, temperature, and salt concentration), sample stability, and the availability of sufficient fruit tissue26,27. This paper presents a protocol for blackcurrant VOCs isolated by HS-SPME and analyzed by gas chromatography coupled with an ion trap mass spectrometer. A balance between the quantity of plant material, sample stability, and duration of extraction and chromatography was achieved to be able to process high numbers of blackcurrant samples, some of them presented in this study. In particular, VOC profiles and/or chromatograms of five cultivars (&#39;Andega&#39;, &#39;Ben Tron&#39;, &#39;Ben Gairn&#39;, &#39;Ben Tirran&#39;, and &#39;Tihope&#39;) will be presented and discussed as example data. Furthermore, the same protocol has been successfully put into practice for VOC measurement in other fruit berry species such as strawberry (Fragaria x ananassa), raspberry (Rubusidaeus), and blueberry (Vaccinium spp.).

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>10 mL screw top headspace vials</td>
			<td>Thermo Scientific</td>
			<td>10-HSV</td>
			<td></td>
		</tr>
		<tr>
			<td>18 mm screw cap Silicone/PTFE</td>
			<td>Thermo Scientific</td>
			<td>18-MSC</td>
			<td></td>
		</tr>
		<tr>
			<td>5 mL Tube with HDPE screw cap</td>
			<td>VWR</td>
			<td>216-0153</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Thermo Scientific</td>
			<td>75002415</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol for HPLC</td>
			<td>Merck</td>
			<td>34860-1L-R</td>
			<td></td>
		</tr>
		<tr>
			<td>N-pentadecane (D32, 98%)</td>
			<td>Cambridge Isotope Laboratories</td>
			<td>DLM-1283-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium chloride</td>
			<td>Merck</td>
			<td>S9888</td>
			<td></td>
		</tr>
		<tr>
			<td>SPME fiber PDMS/DVB</td>
			<td>Merck</td>
			<td>57345-U</td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless grinding jars for TissueLyser</td>
			<td>Qiagen</td>
			<td>69985</td>
			<td></td>
		</tr>
		<tr>
			<td>TissueLyser II</td>
			<td>Qiagen</td>
			<td>85300</td>
			<td>Can be subsituted by mortar and pestle or cryogenic mill</td>
		</tr>
		<tr>
			<td>Trace GC gas chromatograph-ITQ900 ion trap mass spectrometer</td>
			<td>Thermo Scientific</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Triplus RSH autosampler with automated SPME device</td>
			<td>Thermo Scientific</td>
			<td>1R77010-0450</td>
			<td></td>
		</tr>
		<tr>
			<td>Water for HPLC</td>
			<td>Merck</td>
			<td>270733-1L</td>
			<td></td>
		</tr>
		<tr>
			<td>Xcalibur 4.2 SP1</td>
			<td>Thermo Scientific</td>
			<td></td>
			<td>software</td>
		</tr>
	</tbody>
</table>

profiling volatile compounds in blackcurrant fruit using headspace solid-phase microextraction coupled to gas chromatography-mass spectrometry

There is an increasing interest in measuring volatile organic compounds (VOCs) emitted by ripe fruits for the purpose of breeding varieties or cultivars with enhanced organoleptic characteristics and thus, to increase consumer acceptance. High-throughput metabolomic platforms have been recently developed to quantify a wide range of metabolites in different plant tissues, including key compounds responsible for fruit taste and aroma quality (volatilomics). A method using headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS) is described here for the identification and quantification of VOCs emitted by ripe blackcurrant fruits, a berry highly appreciated for its flavor and health benefits.
Ripe fruits of blackcurrant plants (Ribes nigrum) were harvested and directly frozen in liquid nitrogen. After tissue homogenization to produce a fine powder, samples were thawed and immediately mixed with sodium chloride solution. Following centrifugation, the supernatant was transferred into a headspace glass vial containing sodium chloride. VOCs were then extracted using a solid-phase microextraction (SPME) fiber and a gas chromatograph coupled to an ion trap mass spectrometer. Volatile quantification was performed on the resulting ion chromatograms by integrating peak area, using a specific m/z ion for each VOC. Correct VOC annotation was confirmed by comparing retention times and mass spectra of pure commercial standards run under the same conditions as the samples. More than 60 VOCs were identified in ripe blackcurrant fruits grown in contrasting European locations. Among the identified VOCs, key aroma compounds, such as terpenoids and C6 volatiles, can be used as biomarkers for blackcurrant fruit quality. In addition, advantages and disadvantages of the method are discussed, including prospective improvements. Furthermore, the use of controls for batch correction and minimization of drift intensity have been emphasized.

High-throughput VOC profiling in a large set of fruit crops grown under different conditions or locations or belonging to distinct genotypes is necessary for accurate aroma phenotyping. Here, a fast and semi-automated HS-SPME/GC-MS platform for relative VOC quantification in blackcurrant cultivars is presented. VOC detection and identification were based on a library that was developed to profile berry fruit species (Table 1). A typical ripe blackcurrant fruit volatile profile (total ion chromatogram) ob...

Watch this Scientific Journal Video about Profiling Volatile Compounds in Blackcurrant Fruit using Headspace Solid-Phase Microextraction Coupled to Gas Chromatography-Mass Spectrometry at JoVE.com

Profiling Volatile Compounds in Blackcurrant Fruit using Headspace Solid-Phase Microextraction Coupled to Gas Chromatography-Mass Spectrometry

A headspace solid-phase microextraction-gas-chromatography platform is described here for fast, reliable, and semi-automated volatile identification and quantification in ripe blackcurrant fruits. This technique can be used to increase knowledge about fruit aroma and to select cultivars with enhanced flavor for the purpose of breeding.

There is an increasing interest in measuring volatile organic compounds (VOCs) emitted by ripe fruits for the purpose of breeding varieties or cultivars ...

Research

JoVE Journal

Biochemistry

Sheathless Capillary Electrophoresis&#8211;Mass Spectrometry for Metabolic Profiling of Biological Samples

Sheathless Capillary ElectrophoresisÃƒÆ’Ã‚Â¢ÃƒÂ¢Ã¢â‚¬Å¡Ã‚Â¬ÃƒÂ¢Ã¢â€šÂ¬Ã…â€œMass Spectrometry for Metabolic Profiling of Biological Samples

In metabolomics, a wide range of analytical techniques is used for the global profiling of (endogenous) metabolites in complex samples. In this paper, a ...

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Most proteins act in association with others; hence, it is crucial to characterize these functional units in order to fully understand biological ...

A Tandem Liquid Chromatography&#8211;Mass Spectrometry-based Approach for Metabolite Analysis of Staphylococcus aureus

A Tandem Liquid ChromatographyÃƒÆ’Ã‚Â¢ÃƒÂ¢Ã¢â‚¬Å¡Ã‚Â¬ÃƒÂ¢Ã¢â€šÂ¬Ã…â€œMass Spectrometry-based Approach for Metabolite Analysis of Staphylococcus aureus

In an effort to thwart bacterial pathogens, hosts often limit the availability of nutrients at the site of infection. This limitation can alter the ...

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Many exceptional advances have been made in mass spectrometry (MS)-based proteomics, with particular technical progress in liquid chromatography (LC) ...

Rapid Collection of Floral Fragrance Volatiles using a Headspace Volatile Collection Technique for GC-MS Thermal Desorption Sampling

Fragrances of many flower families have been sampled and the volatiles analyzed. Knowing the compounds that make up the fragrances can be an important ...

Nitropeptide Profiling and Identification Illustrated by Angiotensin II

Protein nitration is one of the most important post-translational modifications (PTM) on tyrosine residues and it can be induced by chemical actions of ...

Absolute Quantification of Cell-Free Protein Synthesis Metabolism by Reversed-Phase Liquid Chromatography-Mass Spectrometry

Cell-free protein synthesis (CFPS) is an emerging technology in systems and synthetic biology for the in vitro production of proteins. However, if CFPS is ...

Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling

Immunoaffinity purification mass spectrometry (IP-MS) has emerged as a robust quantitative method of identifying protein-protein interactions. This ...

Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry

Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry

Lipids are structurally diverse amphipathic molecules that are insoluble in water. Lipids are essential contributors to the organization and function of ...

Untargeted Liquid Chromatography-Mass Spectrometry-Based Metabolomics Analysis of Wheat Grain

Understanding the interactions between genes, the environment and management in agricultural practice could allow more accurate prediction and management ...