Pott Delphine M, Vallarino José G, Osorio Sonia
Departamento de Biología Molecular y Bioquímica, Campus de Teatinos, Instituto de Hortofruticultura Subtropical y Mediterránea "La Mayora", Universidad de Málaga-Consejo Superior de Investigaciones Científicas.
Departamento de Biología Molecular y Bioquímica, Campus de Teatinos, Instituto de Hortofruticultura Subtropical y Mediterránea "La Mayora", Universidad de Málaga-Consejo Superior de Investigaciones Científicas;
J Vis Exp. 2021 Jun 9(172). doi: 10.3791/62421.
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.
为了培育具有增强感官特性从而提高消费者接受度的品种或栽培品种，人们对测量成熟果实释放的挥发性有机化合物（VOCs）的兴趣与日俱增。最近开发了高通量代谢组学平台，以量化不同植物组织中的多种代谢物，包括负责果实味道和香气品质的关键化合物（挥发物组学）。本文描述了一种使用顶空固相微萃取（HS-SPME）结合气相色谱-质谱联用（GC-MS）的方法，用于鉴定和定量成熟黑加仑果实释放的VOCs，黑加仑是一种因其风味和健康益处而备受赞赏的浆果。收获黑加仑植株（黑醋栗）的成熟果实，并直接在液氮中冷冻。组织匀浆制成细粉后，将样品解冻并立即与氯化钠溶液混合。离心后，将上清液转移到含有氯化钠的顶空玻璃瓶中。然后使用固相微萃取（SPME）纤维和与离子阱质谱仪联用的气相色谱仪提取VOCs。通过积分峰面积，使用每种VOC的特定m/z离子，对所得离子色谱图进行挥发性定量。通过比较在与样品相同条件下运行的纯商业标准品的保留时间和质谱，确认了正确的VOC注释。在欧洲不同地点种植的成熟黑加仑果实中鉴定出了60多种VOCs。在鉴定出的VOCs中，关键香气化合物，如萜类化合物和C6挥发物，可作为黑加仑果实品质的生物标志物。此外，还讨论了该方法的优缺点，包括预期的改进。此外，强调了使用对照进行批次校正和最小化漂移强度。
