基于HS-SPME-GC-MS与分子对接技术的3种香型红茶挥发性成分研究

茶叶科学 ›› 2025, Vol. 45 ›› Issue (2): 318-332.

基于HS-SPME-GC-MS与分子对接技术的3种香型红茶挥发性成分研究

张鹏¹, 黄艳², 魏成江¹, 郑志强¹, 吴伟伟¹, 郑昌坤¹, 申卫伟³, 于英杰³, 林馥茗^2,*, 孙威江^1,*

1.福建农林大学园艺学院,福建福州 350002;
2.福建农林大学安溪茶学院,福建泉州 362400;
3.中国茶叶流通协会北京 100801

收稿日期:2024-12-10 修回日期:2025-02-24 出版日期:2025-04-15 发布日期:2025-04-30
通讯作者: *497147822@qq.com;swj8103@126.com
作者简介:张鹏,男,硕士研究生,主要从事茶叶品质化学方面的研究。
基金资助:
中央引导地方科技发展专项（2022L3071）、福建张天福茶叶发展基金会科技创新基金（FJZTF01）、两岸“三茶融合”标准共通试点项目、五指山红茶品质提升关键技术示范与推广（KH200116A）

The Study of Volatile Components in Three Scented Types of Black Tea Based on HS-SPME-GC-MS and Molecular Docking Technology

ZHANG Peng¹, HUANG Yan², WEI Chengjiang¹, ZHENG Zhiqiang¹, WU Weiwei¹, ZHENG Changkun¹, SHEN Weiwei³, YU Yingjie³, LIN Fuming^2,*, SUN Weijiang^1,*

1. College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, China;
2. Anxi College of Tea Science, Fujian Agriculture and Forestry University, Quanzhou 362400, China;
3. China Tea Circulation Association, Beijing 100801, China

Received:2024-12-10 Revised:2025-02-24 Online:2025-04-15 Published:2025-04-30

摘要/Abstract

摘要： 近年来,蜜香型、果香型和草本薄荷香型红茶深受消费者的关注与喜爱,但其香气特征的成因亟需深入研究与解析。本研究采用顶空固相微萃取-气相色谱-质谱联用（HS-SPME-GC-MS）,结合正交偏最小二乘法判别分析（OPLS-DA）的变量投影重要性值（VIP）和相对香气活性值（ROAV）,确认了3种香型红茶的关键挥发性化合物,最终利用分子对接技术探讨关键挥发性化合物与嗅感受体的结合位点和相互作用。结果表明,3种香型红茶的挥发性成分含量存在显著差异,共有13种挥发性化合物被鉴定为导致这些香型差异的关键因素。在蜜香型红茶中,大马士酮、苯甲醛和芳樟醇氧化物Ⅰ是主要贡献挥发性化合物;在果香型红茶中,庚醛、3,6-亚壬基-1-醇、2-庚酮、(E)-柠檬醛和6-甲基-5-庚烯-2-酮起着至关重要的作用;而草本薄荷香型红茶中的清凉感与水杨酸甲酯密切相关。分子对接结果显示,红茶关键挥发性化合物能自发与OR1A1、OR1G1、OR2W1、OR5M3、OR7D4和OR8D1嗅感受体结合,其中OR1A1是感知3种香型特征的关键受体,关键挥发性化合物主要通过激活OR1A1的3个氨基酸残基（TYR258、PHE206和VAL254）而发生氢键和疏水相互作用,从而促进3种香气的展现。本研究揭示了3种香型红茶特征香气形成的原因,为提升红茶风味品质和实现定向加工提供了重要的理论依据。

关键词: 红茶, 独特香型, 挥发性成分, 分子对接, 相对香气活性值

Abstract: In recent years, honey-like, fruity, and herbal mint-scented black teas have attracted considerable consumer attention and preference. However, the underlying mechanisms of their aroma characteristics require further in-depth investigation and analysis. This study employed headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS), combined with variable importance in projection (VIP) and relative odor activity value (ROAV) of orthogonal partial least squares-discriminant analysis (OPLS-DA) to identify key volatile compounds in three scented types of black tea. Molecular docking was then used to explore the binding sites and interactions between the key volatile compounds and olfactory receptors. The results show significant differences in the volatile component contents among the three scented types of black tea, with 13 volatile compounds identified as the critical contributors to these differences. In honey-like black tea, damascenone, benzaldehyde, and linalool oxide I were identified as the major volatile contributors. In fruity-scented black tea, heptanal, 3,6-nonadien-1-ol, 2-heptanone, (E)-citral, and 6-methyl-5-hepten-2-one played pivotal roles. While the cooling sensation in herbal mint-scented black tea was closely associated with methyl salicylate. Molecular docking analysis demonstrates that the key volatile compounds spontaneously bind to olfactory receptors OR1A1, OR1G1, OR2W1, OR5M3, OR7D4, and OR8D1, with OR1A1 identified as the primary receptor for perceiving these aroma characteristics. The binding was facilitated by hydrogen bonding and hydrophobic interactions with three amino acid residues (TYR258, PHE206, and VAL254) of OR1A1, promoting the presentation of the aroma profiles. This study elucidates the mechanisms for the characteristic aroma formations of these three scented types of black tea, providing a theoretical foundation for enhancing black tea flavor quality and achieving targeted processing.

Key words: black tea, unique aroma types, volatile components, molecular docking, relative odor activity value

中图分类号:

S571.1
TS272

张鹏, 黄艳, 魏成江, 郑志强, 吴伟伟, 郑昌坤, 申卫伟, 于英杰, 林馥茗, 孙威江. 基于HS-SPME-GC-MS与分子对接技术的3种香型红茶挥发性成分研究[J]. 茶叶科学, 2025, 45(2): 318-332.

ZHANG Peng, HUANG Yan, WEI Chengjiang, ZHENG Zhiqiang, WU Weiwei, ZHENG Changkun, SHEN Weiwei, YU Yingjie, LIN Fuming, SUN Weijiang. The Study of Volatile Components in Three Scented Types of Black Tea Based on HS-SPME-GC-MS and Molecular Docking Technology[J]. Journal of Tea Science, 2025, 45(2): 318-332.

参考文献

[1] Pan S Y, Nie Q, Tai H C, et al.Tea and tea drinking: China’s outstanding contributions to the mankind[J]. Chinese Medicine, 2022, 17(1): 27. doi: 10.1186/s13020-022-00571-1.
[2] Chen Q C, Zhu Y, Liu Y F, et al.Black tea aroma formation during the fermentation period[J]. Food Chemistry, 2022, 374: 131640. doi:10.1016/j.foodchem.2021.131640.
[3] Sanderson G W, Grahamm H N. Formation of black tea aroma[EB/OL]. American Chemical Society, 2002[2024-11-13]. https://pubs.acs.org/doi/pdf/10.1021/jf60188a007.
[4] Wu Q Y, Zhou Z W, Zhang Y N, et al.Identification of key components responsible for the aromatic quality of Jinmudan black tea by means of molecular sensory science[J]. Foods, 2023, 12(9): 1794. doi: 10.3390/foods12091794.
[5] 欧阳珂, 张成, 廖雪利, 等. 基于感官组学分析玉米香型南川大茶树工夫红茶特征香气[J]. 茶叶科学, 2022, 42(3): 397-408.
Ouyang K, Zhang C, Liao X L, et al.Characterization of the key aroma in corn-scented Congou black tea manufactured from Camellia nanchuanica by sensory omics techniques[J]. Journal of Tea Science, 2022, 42(3): 397-408.
[6] Sun Z C, Lin Y C, Yang H, et al.Characterization of honey-like characteristic aroma compounds in Zunyi black tea and their molecular mechanisms of interaction with olfactory receptors using molecular docking[J]. LWT, 2024, 191: 115640. doi: 10.1016/j.lwt.2023.115640.
[7] Zheng F L, Gan S Y, Zhao X Y, et al.Unraveling the chemosensory attributes of Chinese black teas from different regions using GC-IMS combined with sensory analysis[J]. LWT, 2023, 184: 114988. doi:10.1016/j.lwt.2023.114988.
[8] Liang Y L, Wang Z H, Zhang L Z, et al.Characterization of volatile compounds and identification of key aroma compounds of in different aroma types of Rougui Wuyi rock tea[J]. Food Chemistry, 2024: 139931. doi: 10.1016/j.foodchem.2024.139931.
[9] Liu N F, Shen S S, Huang L F, et al.Revelation of volatile contributions in green teas with different aroma types by GC-MS and GC-IMS[J]. Food Research International, 2023, 169: 112845. doi: 10.1016/j.foodres.2023.112845.
[10] Yang Y Q, Zhu H K, Chen J Y, et al.Characterization of the key aroma compounds in black teas with different aroma types by using gas chromatography electronic nose, gas chromatography-ion mobility spectrometry, and odor activity value analysis[J]. LWT, 2022, 163: 113492. doi: 10.1016/j.lwt.2022.113492.
[11] Pinzi L, Rastelli G.Molecular docking: shifting paradigms in drug discovery[J]. International Journal of Molecular Sciences, 2019, 20(18): 4331. doi: 10.3390/ijms20184331.
[12] Mei S H, Ding J, Chen X M.Identification of differential volatile and non-volatile compounds in coffee leaves prepared from different tea processing steps using HS-SPME/GC-MS and HPLC-Orbitrap-MS/MS and investigation of the binding mechanism of key phytochemicals with olfactory and taste receptors using molecular docking[J]. Food Research International, 2023, 168: 112760. doi: 10.1016/j.foodres.2023.112760.
[13] Xiao Z B, Shen T Y, Ke Q F, et al.Identification of characteristic aroma compounds of Longjing tea and their molecular mechanisms of interaction with olfactory receptors using molecular docking[J]. European Food Research and Technology, 2024, 250(5): 1363-1378.
[14] Zhu J C, Liu X J, Lin Y C, et al.Unraveling the characteristic chestnut aroma compounds in Meitan Cuiya green tea and their interaction mechanisms with broad-spectrum olfactory receptors using molecular docking[J]. LWT, 2024, 194: 115785. doi: 10.1016/j.lwt.2024.115785.
[15] Li H H, Luo L Y, Wang J, et al.Lexicon development and quantitative descriptive analysis of Hunan Fuzhuan brick tea infusion[J]. Food Research International, 2019, 120: 275-284.
[16] Wang Z H, Liang Y L, Gao C X, et al.The flavor characteristics and antioxidant capability of aged Jinhua white tea and the mechanisms of its dynamic evolution during long-term aging[J]. Food Chemistry, 2024, 436: 137705. doi: 10.1016/j.foodchem.2023.137705.
[17] Wang Y J, Huang L F, Deng G J, et al.The shaking and standing processing improve the aroma quality of summer black tea[J]. Food Chemistry, 2024, 454: 139772. doi: 10.1016/j.foodchem.2024.139772.
[18] Xu Y J, Liu Y Q, Yang J H, et al.Manufacturing process differences give Keemun black teas their distinctive aromas[J]. Food Chemistry: X, 2023, 19: 100865. doi: 10.1016/j.fochx.2023.100865.
[19] Liao X L, Yan J N, Wang B, et al.Identification of key odorants responsible for cooked corn-like aroma of green teas made by tea cultivar ‘Zhonghuang 1’[J]. Food Research International, 2020, 136: 109355. doi: 10.1016/j.foodres.2020.109355.
[20] Huang S Y, Tao L L, Xu L L, et al.Discrepancy on the flavor compound affect the quality of Taiping Houkui tea from different production regions[J]. Food Chemistry: X, 2024, 23: 101547. doi: 10.1016/j.fochx.2024.101547.
[21] Yin P, Kong Y S, Liu P P, et al.A critical review of key odorants in green tea: identification and biochemical formation pathway[J]. Trends in Food Science & Technology, 2022, 129: 221-232.
[22] Zhu J C, Niu Y W, Xiao Z B.Characterization of the key aroma compounds in Laoshan green teas by application of odour activity value (OAV), gas chromatography-mass spectrometry-olfactometry (GC-MS-O) and comprehensive two-dimensional gas chromatography mass spectrometry (GC × GC-qMS)[J]. Food Chemistry, 2021, 339: 128136. doi: 10.1016/j.foodchem.2020.128136.
[23] 侯智炜, 吕永铭, 马宽, 等. 不同茶树品种的径山茶挥发性成分差异研究[J]. 茶叶科学, 2024, 44(5): 747-762.
Hou Z W, Lü Y M, Ma K, et al.Study on the differences in volatile components of Jingxian tea from different tea cultivars[J]. Journal of Tea Science, 2024, 44(5): 747-762.
[24] Wang Z H, Liang Y L, Wu W W, et al.The effect of different drying temperatures on flavonoid glycosides in white tea: a targeted metabolomics, molecular docking, and simulated reaction study[J]. Food Research International, 2024, 190: 114634. doi: 10.1016/j.foodres.2024.114634.
[25] Wang B Y, Chen H M, Qu F F, et al.Identification of aroma-active components in black teas produced by six Chinese tea cultivars in high-latitude region by GC-MS and GC-O analysis[J]. European Food Research and Technology, 2022, 248(3): 647-657.
[26] Bezman Y, Bilkis I, Winterhalter P, et al.Thermal oxidation of 9'-cis-neoxanthin in a model system containing peroxyacetic acid leads to the potent odorant β-damascenone[J]. Journal of Agricultural and Food Chemistry, 2005, 53(23): 9199-9206.
[27] Zhai X T, Zhang L, Granvogl M, et al.Flavor of tea (Camellia sinensis): a review on odorants and analytical techniques[J]. Comprehensive Reviews in Food Science and Food Safety, 2022, 21(5): 3867-3909.
[28] Ma J Q, Wang Y J, Li J Y, et al.Aroma formation mechanism by the drying step during Congou black tea processing: analyses by HP-SPME and SAFE with GC-MS[J]. LWT, 2024, 198: 116019. doi: 10.1016/j.lwt.2024.116019.
[29] Mahmoud M A A, Kılıç-Büyükkurt Ö, Aboul Fotouh M M, et al. Aroma active compounds of honey: analysis with GC-MS, GC-O, and molecular sensory techniques[J]. Journal of Food Composition and Analysis, 2024, 134: 106545. doi: 10.1016/j.jfca.2024.106545.
[30] Wang X Q, Zeng L T, Liao Y Y, et al.An alternative pathway for the formation of aromatic aroma compounds derived from l-phenylalanine via phenylpyruvic acid in tea (Camellia sinensis (L.) O. Kuntze) leaves[J]. Food Chemistry, 2019, 270: 17-24.
[31] Costa A C, Garruti D S, Madruga M S.The power of odour volatiles from unifloral melipona honey evaluated by gas chromatography-olfactometry osme techniques[J]. Journal of the Science of Food and Agriculture, 2019, 99(9): 4493-4497.
[32] Duru M E, Taş M, Çayan F, et al.Characterization of volatile compounds of Turkish pine honeys from different regions and classification with chemometric studies[J]. European Food Research and Technology, 2021, 247(10): 2533-2544.
[33] Shao C Y, Zhang Y, Lü H P, et al.Aromatic profiles and enantiomeric distributions of chiral odorants in baked green teas with different picking tenderness[J]. Food Chemistry, 2022, 388: 132969. doi: 10.1016/j.foodchem.2022.132969.
[34] Demyttenaere J C R, Willemen H M. Biotransformation of linalool to furanoid and pyranoid linalool oxides by Aspergillus niger[J]. Phytochemistry, 1998, 47(6): 1029-1036.
[35] Makowicz E, Jasicka-Misiak I, Teper D, et al.Botanical origin authentication of polish phacelia honey using the combination of volatile fraction profiling by HS-SPME and lipophilic fraction profiling by HPTLC[J]. Chromatographia, 2019, 82(10): 1541-1553.
[36] Jerković I, Kuś P M.Headspace solid-phase microextraction and ultrasonic extraction with the solvent sequences in chemical profiling of Allium ursinum L. honey[J]. Molecules, 2017, 22(11): 1909. doi: 10.3390/molecules22111909.
[37] Pattamayutanon P, Angeli S, Thakeow P, et al.Volatile organic compounds of Thai honeys produced from several floral sources by different honey bee species[J]. PLoS ONE, 2017, 12(2): e0172099. doi: 10.1371/journal.pone.0172099.
[38] Wang Q W, Xie J L, Wang L L, et al.Comprehensive investigation on the dynamic changes of volatile metabolites in fresh scent green tea during processing by GC-E-Nose, GC-MS, and GC × GC-TOFMS[J]. Food Research International, 2024, 187: 114330. doi: 10.1016/j.foodres.2024.114330.
[39] Wang J T, Zhu Y, Shi J, et al.Discrimination and identification of aroma profiles and characterized odorants in citrus blend black tea with different citrus species[J]. Molecules, 2020, 25(18): 4208. doi: 10.3390/molecules25184208.
[40] Wu Z R, Jiao Y F, Jiang X F, et al.Effects of sun withering degree on black tea quality revealed via non-targeted metabolomics[J]. Foods, 2023, 12(12): 2430. doi: 10.3390/foods12122430.
[41] Deng H L, Chen S S, Zhou Z W, et al.Transcriptome analysis reveals the effect of short-term sunlight on aroma metabolism in postharvest leaves of oolong tea (Camellia sinensis)[J]. Food Research International, 2020, 137: 109347. doi: 10.1016/j.foodres.2020.109347.
[42] Huang W J, Fang S M, Wang J, et al.Sensomics analysis of the effect of the withering method on the aroma components of Keemun black tea[J]. Food Chemistry, 2022, 395: 133549. doi: 10.1016/j.foodchem.2022.133549.
[43] Duan Y, Yu M G, Raza J, et al.Effect of roasting time on aroma quality of Shuixian Wuyi Rock Tea (Camellia sinensis)[J]. Journal of Food Composition and Analysis, 2024, 135: 106662. doi: 10.1016/j.jfca.2024.106662.
[44] Wang D L, Liu Z B, Lan X Y, et al.Unveiling the aromatic intricacies of Wuyi rock tea: a comparative study on sensory attributes and odor-active compounds of Rougui and Shuixian varieties[J]. Food Chemistry, 2024, 435: 137470. doi: 10.1016/j.foodchem.2023.137470.
[45] Wang J, Li M R, Wang H, et al.Decoding the specific roasty aroma Wuyi rock tea (Camellia sinensis: Dahongpao) by the sensomics approach[J]. Journal of Agricultural and Food Chemistry, 2022, 70(34): 10571-10583.
[46] Li X, Zhang L P, Zhang L, et al.Methyl salicylate enhances flavonoid biosynthesis in tea leaves by stimulating the phenylpropanoid pathway[J]. Molecules, 2019, 24(2): 362. doi: 10.3390/molecules24020362.
[47] Kang S Y, Yan H, Zhu Y, et al.Identification and quantification of key odorants in the world’s four most famous black teas[J]. Food Research International, 2019, 121: 73-83.
[48] Zhang D, Huang Y J, Fan X, et al.Effects of solid-state fermentation with Aspergillus cristatus (MK346334) on the dynamics changes in the chemical and flavor profile of dark tea by HS-SPME-GC-MS, HS-GC-IMS and electronic nose[J]. Food Chemistry, 2024, 455: 139864. doi: 10.1016/j.foodchem.2024.139864.
[49] Xiao Z B, Shen T Y, Niu Y W, et al.Unraveling the characteristic aroma compounds in Longjing tea and their interaction mechanisms with s-curve and broad-spectrum olfactory receptors using molecular docking[J]. Food Bioscience, 2024, 60: 104423. doi: 10.1016/j.fbio.2024.
104423.
[50] Zhu J C, Chen Y Q, Liu X J, et al.Exploring the molecular mechanism of the interaction between characteristic aroma compounds in Longjing tea using a combination of sensory and theoretical perspectives: with a focus on linalool and methyl jasmonate on olfactory receptor of OR52D1[J]. Industrial Crops and Products, 2025, 224: 120314. doi: 10.1016/j.indcrop.2024.120314.
[51] Zeng S T, Zhang L L, Li P, et al.Molecular mechanisms of caramel-like odorant-olfactory receptor interactions based on a computational chemistry approach[J]. Food Research International, 2023, 171: 113063. doi: 10.1016/j.foodres.2023.113063.