超高效液相色谱-四级杆飞行时间质谱结合Progenesis QI 分析不同品种橙的指纹图谱及其差异性代谢物

doi:10.3864/j.issn.0578-1752.2018.13.010

摘要/Abstract

摘要： 【目的】建立分析不同品种橙之间差异性代谢物的方法，找出各品种橙的特征代谢物，服务于橙的代谢组学研究，为橙果实和橙汁的鉴别提供参考。【方法】选取相同产地8个不同的橙品种，取其果皮的甲醇提取物，使用超高效液相色谱-四级杆飞行时间质谱（UPLC-QToF-MS）得到对应的指纹图谱，结合组学软件Progenesis QI，通过主成分分析、偏最小二乘判别分析和变量变化趋势图等进行综合评价，筛选出各品种间的差异性代谢物。根据精确分子量、二级碎片、标准品以及数据库和相关文献信息，对差异性代谢物进行鉴定。【结果】通过UPLC-QToF-MS指纹图谱得出8个橙品种的代谢物存在明显差异，使用Progenesis QI建立的流程筛选得到各品种正、负离子模式下的差异性代谢物，包含保留时间、分子式、二级碎片等信息。鉴定出17种差异代谢物，其中3-羟基-5,7,3',4'-四甲氧基黄酮和木犀草素-7-O-新橙皮糖苷是血橙区别于其他7种橙的特征代谢物，橙皮苷是S26锦橙的代谢标志物，芸香柚皮苷-4'-O-葡萄糖苷和异橙黄酮可以作为8045甜橙的特征代谢物。【结论】基于UPLC-QToF-MS和Progenesis QI建立的方法可以筛选出8个品种橙果实的差异性代谢物，鉴定了其中的17种物质，分别对应不同品种。该研究方法和技术路线具有通用性，适用于不同水果、不同产地、不同成熟度等因素引起的样品差异分析。

关键词: 超高效液相色谱-四级杆飞行时间质谱, 橙, Progenesis QI, 代谢物

Abstract: 【Objective】The objective of this study is to establish an efficient method to investigate the differences between different varieties of oranges, and to find their differential metabolites, which will serve the citrus metabolomics and provide reference for the identification of oranges and orange juice. 【Method】 8 orange cultivars from the same area were chosen to obtain the methanol extracts of their peels, and then based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QToF-MS), the LC-MS fingerprints of the 8 oranges were obtained with obvious differences. Combined with the software Progenesis QI, the differential metabolites were screened in positive and negative modes through principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) as well as loadings plots and VIP (variable importance for projection) values. Furthermore, the differential metabolites were identified based on retention times, accurate mass, MS/MS fragments, reference standards and public databases.【Result】Based on the established method, differential metabolites of the 8 orange cultivars were screened. 17 compounds had been identified and among them, and 6 compounds were confirmed by comparison with their commercial standards. 3-Hydroxy-5,7,3',4'-tetramethoxyflavone and luteolin-7-O-neohesperidoside were the characteristic metabolites of Xuecheng. Hesperidin was the marker compound of S26 Jincheng. Narirutin-4'-glucoside and Isosinensetin could act as the characteristic metabolites of 8045 Tiancheng. 【Conclusion】 The method was versatile and suitable for the analysis of sample differences caused by varieties, geographical origins and maturity.

Key words: ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry, orange, Progenesis QI, metabolites

赵希娟，赵无疾，许华超. 超高效液相色谱-四级杆飞行时间质谱结合Progenesis QI 分析不同品种橙的指纹图谱及其差异性代谢物[J]. 中国农业科学, 2018, 51(13): 2551-2560.

ZHAO XiJuan, ZHAO WuJi, XU HuaChao. Analysis of the Fingerprints of Different Orange Varieties and Their Differential Metabolites Based on Ultra-Performance Liquid Chromatography Coupled with Quadrupole Time-of-Flight Mass Spectrometry and Progenesis QI[J]. Scientia Agricultura Sinica, 2018, 51(13): 2551-2560.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2018.13.010

https://www.chinaagrisci.com/CN/Y2018/V51/I13/2551

参考文献

[1] 刘贤青, 张红艳. HPLC-Q-TOF/MS分析脐橙果实中的类黄酮. 植物科学学报, 2014, 32(6): 638-644.

LIU X Q, ZHANG H Y. Determination of flavonoids from Navel Orange (Citrus sinensis) fruits by HPLC-Q-TOF/MS. Plant Science Journal, 2014, 32(6): 638-644. (in Chinese)

[2] 从彦丽, 彭梦雪, 刘冬, 孙海燕, 唐旭蔚. 柑橘在体外模拟胃肠消化过程中总多酚、总黄酮及总抗氧化活性的变化规律. 食品科学, 2016, 37(17): 96-103.

CONG Y L, PENG M X, LIU D, SUN H Y, TANG X W. Changes in total polyphenols, total flavonoids and antioxidant activity of Citrus during in vitro gastrointestinal digestion process. Food Science, 2016, 37(17): 96-103. (in Chinese)

[3] JANDRI? Z, CANNAVAN A. An investigative study on differentiation of citrus fruit/fruit juices by UPLC-QToF MS and chemometrics. Food Control, 2017, 72: 173-180.

[4] Muntean E. Simultaneous carbohydrate chromatography and unsuppressed ion chromatography in detecting fruit juices adulteration. Chromatographia, 2010, 71(1): 69-74.

[5] PROTTI M, VALLE F, POLI F, RAGGIA M A, MERCOLINI L. Bioactive molecules as authenticity markers of Italian Chinotto (Citrus×myrtifolia) fruits and beverages. Journal of Pharmaceutical and Biomedical Analysis, 2015, 104: 75-80.

[6] LIM S, LEE J G, LEE E J. Comparison of fruit quality and GC–MS-based metabolite profiling of kiwifruit ‘Jecy green’: Natural and exogenous ethylene-induced ripening. Food Chemistry, 2017, 234: 81-92.

[7] BORGES G, CROZIER A. HPLC–PDA–MS fingerprinting to assess the authenticity of pomegranate beverages. Food Chemistry, 2012, 135: 1863-1867.

[8] 陈永刚, 林励. 柑橘属常用中药黄酮类成分HPLC指纹图谱研究与比较. 中国中药杂志, 2011, 36(19): 2660-2665.

CHEN Y G, LIN L. Study and comparison on HPLC fingerprints of flavonoids of frequently used Chinese materia medica in citrus. China Journal of Chinese Materia Medica, 2011, 36(19): 2660-2665. (in Chinese)

[9] 郑洁, 赵其阳, 张耀海, 焦必宁. 超高效液相色谱法同时测定柑橘中主要酚酸和类黄酮物质. 中国农业科学, 2014, 47(23): 4706-4717.

ZHENG J, ZHAO Q Y, ZHANG Y H, JIAO B N. Simultaneous determination of main flavonoids and phenolic acids in citrus fruit by ultra performance liquid chromatography. Scientia Agricultura Sinica, 2014, 47(23): 4706-4717. (in Chinese)

[10] 刘贤青, 涂虹, 王守创, 张红艳, 罗杰, 徐娟. 不同类型柑橘果实汁胞中类黄酮的液相色谱质谱联用分析. 植物生理学报, 2016, 52(5): 762-770.

LIU X Q, TU H, WANG S C, ZHANG H Y, LUO J, XU J. Flavonoid composition of citrus juice sacs determined by high-performance liquid chromatography coupled with tandem electrospray ionization mass spectrometry. Plant Physiology Journal, 2016, 52(5): 762-770. (in Chinese)

[11] SANDÍN-ESPAÑ P, MATEO-MIRANDA M, LÓPEZ-GOTI C, DE CAL A, ALONSO-PRADOS J L. Development of a rapid and direct method for the determination of organic acids in peach fruit using LC-ESI-MS. Food Chemistry, 2016, 192(Sul C): 268-273.

[12] AVULA B, SAGI S, WANG Y H, WANG M, GAFNER S, MANTHEY J A, KHAN I A. Liquid chromatography-electrospray ionization mass spectrometry method analysis of limonoid aglycones, limonoid acids, limonoid glucosides and flavonoids from grapefruit seeds, other citrus species and dietary supplements. Planta Medica, 2016, 82: 1058-1069.

[13] LÓPEZ-COBO A, GÓMEZ-CARAVACA A M, PASINI F, CABONI M F, SEGURA-CARRETERO A, FERNÁNDEZ-GUTIÉRREZ A. HPLC-DAD-ESI-QTOF-MS and HPLC-FLD-MS as valuable tools for the determination of phenolic and other polar compounds in the edible part and by-products of avocado. LWT-Food Science and Technology, 2016, 73: 505-513.

[14] 陈林林, 米强, 辛嘉英. 柑橘皮精油成分分析及抑菌活性研究. 食品科学, 2010, 31(17): 25-28.

CHEN L L, MI Q, XIN J Y. Composition analysis and antibacterial activity of the essential oil from citrus peel. Food Science, 2010, 31(17): 25-28. (in Chinese)

[15] XING T T, ZHAO X J, ZHANG Y D, LI Y F. Fast separation and sensitive quantitation of polymethoxylated flavonoids in the peels of Citrus using UPLC-Q-TOF-MS. Journal of Agricultural and Food Chemistry, 2017, 65(12): 2615-2627.

[16] YANG Y, ZHAO X J, PAN Y, ZHOU Z. Identification of the chemical compositions of Ponkan peel by ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Analytical Methods, 2016, 8(4): 893-903.

[17] JANDRI? Z, ROBERTS D, RATHOR M N, ABRAHIM A, IALAM M, CANNAVAN A. Assessment of fruit juice authenticity using UPLC–QToF MS: A metabolomics approach. Food Chemistry, 2014, 148: 7-17.

[18] 常玉玮, 王国栋. LC-MS在植物代谢组学分析中的应用. 生命科学, 2015, 27(8): 978-985.

CHANG Y W, WANG G D. Application of LC-MS in plant metabolomics. Chinese Bulletin of Life Sciences, 2015, 27(8): 978-985. (in Chinese)

[19] GUO J, YUE T, YUAN Y, WANG Y. Chemometric classification of apple juices according to variety and geographical origin based on polyphenolic profiles. Journal of Agricultural and Food Chemistry, 2013, 61(28): 6949-6963.

[20] JANDRI? Z, ISLAM M, SINGH D K, CANNAVAN A. Authentication of Indian citrus fruit/fruit juices by untargeted and targeted metabolomics. Food Control, 2017, 72: 181-188.

[21] ZHANG J, YANG W, LI S, YAO S, QI P, YANG Z, FENG Z, HOU J, CAI L, YANG M, WU W, GUO D-A. An intelligentized strategy for endogenous small molecules characterization and quality evaluation of earthworm from two geographic origins by ultra-high performance HILIC/QTOF MS^E and Progenesis QI. Analytical and Bioanalytical

Chemistry, 2016, 408(14): 3881-3890.

[22] YAO C, YANG W, ZHANG J, QIU S, CHEN M, SHI X, PAN H, WU W, GUO D. UHPLC-Q-TOF-MS-based metabolomics approach to compare the saponin compositions of Xueshuantong injection and Xuesaitong injection. Journal of Separation Science, 2017, 40(4): 834-841.

[23] GIKA H G, THEODORIDIS G A, WINGATE J E, WILSON I D. Within-Day reproducibility of an HPLC-MS-based method for metabonomic analysis: application to human urine. Journal of Proteome Research, 2007, 6(8): 3291-3303.

[24] CHEN S, KONG H, LU X, LI Y, YIN P, ZENG Z, XU G. Pseudotargeted metabolomics method and its application in serum biomarker discovery for hepatocellular carcinoma based on ultra high-performance liquid chromatography/triple quadrupole mass spectrometry. Analytical Chemistry, 2013, 85(17): 8326-8333.

[25] CUYCKENS F, CLAEYS M. Mass spectrometry in the structural analysis of flavonoids. Journal of Mass Spectrometry, 2004, 39(1): 1-15.

[26] YE X, CAO D, ZHAO X, SONG F, HUANG Q, FAN G, WU F. Chemical fingerprint and metabolic profile analysis of Citrus reticulate ‘Chachi’ decoction by HPLC-PDA-IT-MSn and HPLC- Quadrupole-Orbitrap-MS method. Journal of Chromatography B, 2014, 970(Sul C): 108-120.

[27] KIM H G, KIM G-S, LEE J H, PARK S, JEONG W Y, KIM Y-H, KIM J H, KIM S T, CHO Y A, LEE W S, LEE S J, JIN J S, SHIN S C. Determination of the change of flavonoid components as the defence materials of Citrus unshiu Marc. fruit peel against Penicillium digitatum by liquid chromatography coupled with tandem mass spectrometry. Food Chemistry, 2011, 128(1): 49-54.

[28] JAYAPRAKASHA G K, DANDEKAR D V, TICHY S E, PATIL B S. Simultaneous separation and identification of limonoids from citrus using liquid chromatography-collision-induced dissociation mass spectra. Journal of Separation Science, 2011, 34(1): 2-10.

[29] WANG S, YANG C, TU H, ZHOU J, LIU X, CHENG Y, LUO J, DENG X, ZHANG H, XU J. Characterization and metabolic diversity of flavonoids in Citrus species. Scientific Reports, 2017, 7(1): 10549.

[30] STANDER M A, KÜHN W, HITEN N F. Survey of South African fruit juices using a fast screening HILIC-MS method. Food Additives & Contaminants: Part A, 2013, 30(9): 1473-1484.