Identification of Antioxidant Components and Tyrosinase Specific Inhibitors from <i>Osmanthus fragrans</i> Flower by Using Online UPLC-ABTS<sup>+</sup>·-assay and UF- LC-MS Technology

ZHOU Huiji; LI Tingzhao; LI Bo

doi:10.13386/j.issn1002-0306.2021070130

Volume 43 Issue 7

Mar. 2022

Turn off MathJax

Article Contents

Abstract

References

Science and Technology of Food Industry > 2022 > 43(7): 67-79. > DOI: 10.13386/j.issn1002-0306.2021070130

ZHOU Huiji, LI Tingzhao, LI Bo. Identification of Antioxidant Components and Tyrosinase Specific Inhibitors from Osmanthus fragrans Flower by Using Online UPLC-ABTS⁺·-assay and UF- LC-MS Technology[J]. Science and Technology of Food Industry, 2022, 43(7): 67−79. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021070130.

Citation:

PDF (7089 KB)

Identification of Antioxidant Components and Tyrosinase Specific Inhibitors from Osmanthus fragrans Flower by Using Online UPLC-ABTS⁺·-assay and UF- LC-MS Technology

ZHOU Huiji^1,,
LI Tingzhao^{1, 2},
LI Bo^{1, 2, ,}

1.
Amway (Shanghai) Technology Development Co., Ltd., Shanghai 201203, China
2.
Amway (China) Botanical R&D Center, Wuxi 214145, China)

More Information

Received Date: July 12, 2021
Available Online: February 10, 2022

Graphical Abstract

Abstract

Abstract

Objective: To investigate and analyze antioxidant activities and tyrosinase inhibition of the aqueous extract of Osmanthus fragrans flower (O. fragrans), as well as its active ingredients. Methods: In this study, antioxidant activities of extract of O. fragrans were carried out by using DPPH·, ABTS⁺· radical scavenging assays and total antioxidant activity (FRAP). Qualitative analysis of major active components was performed by UPLC-PDA-QDa-ABTS⁺· online method and UF-LC-MS method were used to qualitatively identify the antioxidant components and tyrosinase inhibitors from O. fragrans extract respectively. Oxidative damage model of skin fibroblasts induced by H₂O₂ was used to evaluate the protective effect of oxidative stress of O. fragrans aqueous extract and its chemical components. Results: Aqueous extract of O. fragrans had good scavenging effect on DPPH· and ABTS⁺· free radical in vitro, and its EC₅₀ values of free radical scavenging effect on DPPH· and ABTS⁺· were 37.66, 38.32 μg/mL, respectively. When the concentration of O. fragrans extract reached 2 mg/mL, FRAP value could be as high as 3.17. The EC₅₀ of tyrosinase diphenoloxidase inhibition of O. fragrans extract was 595.9 μg/mL. Through analysis of UPLC-Triple-TOF/MS, a total of 28 chemical compositions were preliminarily identified. Five antioxidant active compounds which also greatly combined with tyrosinase were screened out from O. fragrans extract by simultaneously using UPLC-PDA-QDa-ABTS⁺· online method and UF-LC-MS method. Among them, the main active ingredient was verbascoside. Antioxidant and tyrosinase assay showed that verbascoside had significant antioxidant activity and tyrosinase inhibitory activity. EC₅₀ values of verbascoside of scavenging effect on DPPH· and ABTS⁺· were 10.27, 14.96 μg/mL, respectively. When the concentration of verbascoside was 1 mg/mL, FRAP value could be up to 4.22. The EC₅₀ of tyrosinase monophenoloxidase inhibition of verbascoside was 477.5 μg/mL, but with little diphenoloxidase inhibitiory effect. The oxidative damage model induced by H₂O₂ was used to further verify the significant protective effects of O. fragrans extract and verbascoside on oxidative damage of skin fibroblasts. Conclusion: The aqueous extract of O. fragrans and its main active component, verbascoside, had good antioxidant activity, tyrosinase inhibitory activity and oxidative stress protection ability.
- Osmanthus fragrans Lour.,
- verbascoside,
- UPLC-ABTS⁺·,
- online antioxidation,
- UF-LC-MS,
- tyrosinase inhibitors

FullText(HTML)

References (50)

References

[1]	施婷婷, 杨秀莲, 王良桂. 3个桂花品种花香组分动态特征及花被片结构解剖学观测[J]. 南京林业大学学报:自然科学版,2020,44(4):12−20. [SHI Tingting, YANG Xiulian, WANG Lianggui. Observation on the dynamic characteristics of floral components of three Osmanthus fragrans varieties and the structural anatomy of tepals[J]. Journal of Nanjing Forestry University:Natural Science Edition,2020,44(4):12−20.
[2]	薛锦赢, 殷佳雅, 黄秀玉, 等. 发酵型桂花酒的工艺优化[J]. 食品研究与开发,2021,42(10):6. [XUE Jinying, YIN Jiaya, HUANG Xiuyu, et al. Process optimization of fermented osmanthus wine[J]. Food Research and Development,2021,42(10):6.
[3]	李振兴, 白杨. 纷纷加码"中国味" 百事可乐和可口可乐的竞争再升级[J]. 中国食品,2020,794(10):76−77. [LI Zhengxing, BAI Yang. The competition between Pepsi and Coca-Cola is escalating[J]. China Food,2020,794(10):76−77.
[4]	刘有停, 张卫红, 周桐, 等. 桂花提取液在抗皮肤老化方面的功效研究[J]. 香料香精化妆品,2019,173(2):29−32. [LIU Youting, ZHANG Weihong, ZHOU Tong, et al. Study on the efficacy of Osmanthus fragrans extract in anti-aging skin[J]. Fragrance, Fragrance and Cosmetics,2019,173(2):29−32.
[5]	赵东, 袁志彬, 孙琳, 等. 桂花及桂花酒的研究进展[J]. 酿酒科技,2017(1):90−94. [ZHAO Dong, YUAN Zhibin, SUN Lin, et al. Research progress of Osmanthus and Osmanthus wine[J]. Wine Science and Technology,2017(1):90−94.
[6]	宋晓芳, 范宝磊, 曾祥玲, 等. HPLC-MS/MS法同时测定不同桂花品种中7个多酚类活性成分的含量[J]. 药物分析杂志,2019,39(10):1811−1820. [SONG Xiaofang, FANG Baolei, ZENG Xiangling, et al. HPLC-MS/MS simultaneous determination of the contents of 7 polyphenolic active components in different Osmanthus species[J]. Journal of Journal of Pharmaceutical Analysis,2019,39(10):1811−1820.
[7]	丁立新, 李焕, 范宝俭, 等. HPLC法测定不同产地桂花中红景天苷和毛蕊花糖苷的含量[J]. 药物分析杂志,2013,33(5):894−897. [DING Lixin, LI Huan, FAN Baojian, et al. Determination of salidroside and verbascoside in Osmanthus fragrans from different origins by HPLC[J]. Journal of Pharmaceutical Analysis,2013,33(5):894−897.
[8]	SHI Yunshi, PENG Mijun, ZHANG Yuping, et al. Combination of preparative HPLC and HSCCC methods to separate phosphodiesterase inhibitors from Eucommia ulmoides bark guided by ultrafiltration-based ligand screening[J]. Analytical & Bioanalytical Chemistry,2013,405(12):4213−4223.
[9]	AYA A K, MURAT K, et al. Antioxidant components of Viburnum opulus L. determined by on-line HPLC-UV-ABTS radical scavenging and LC-UV-ESI-MS methods[J]. Food Chemistry,2015,175:106−114. doi: 10.1016/j.foodchem.2014.11.085
[10]	YANG Yinjun, SUN Xinguang, LIU Jinjun, et al. Quantitative and qualitative analysis of flavonoids and phenolic acids in snow chrysanthemum (Coreopsis tinctoria Nutt.) by HPLC-DAD and UPLC-ESI-QTOF-MS[J]. Molecules,2016,21(10):1307. doi: 10.3390/molecules21101307
[11]	甘小娜, 彭博, 李廷钊, 等. UPLC-PDA-MS-ABTS阳离子自由基在线分析藤茶的抗氧化活性成分[J]. 食品科学,2020,41(18):254−259. [GAN Xiaona, PENG Bo, LI Tingzhao, et al. Cationic free radical online analysis of the antioxidant active components of vine tea[J]. Food Science,2020,41(18):254−259.
[12]	RANI M P, PADMAKUMARI K P. HPTLC and reverse phase HPLC methods for the simultaneous quantification and in vitro screening of antioxidant potential of isolated sesquiterpenoids from the rhizomes of Cyperus rotundus[J]. Journal of Chromatography B Analytical Technologies in the Biomedical & Life Sciences,2012,904:22−28.
[13]	CALDERN M A I. Development of an ultrafiltration-liquid chromatography/mass spectrometry (UF-LC/MS) based ligand-binding assay and an LC/MS based functional assay for Mycobacterium tuberculosis shikimate kinase[J]. Analytical Chemistry,2010,82(9):3616−3621. doi: 10.1021/ac902849g
[14]	S FANG, L SHEN, Z MA. Screening for ligands of human aromatase from mulberry (Mori alba L.) leaf by using high-performance liquid chromatography/tandem mass spectrometry[J]. Food Chemistry,2011,126(3):1337−1343. doi: 10.1016/j.foodchem.2010.11.096
[15]	CHOI Y, JERMIHOV K, NAM S J, et al. Correction to screening natural products for inhibitors of quinone reductase-2 using ultrafiltration LC–MS[J]. Analytical Chemistry,2011,83(11):4325−4325. doi: 10.1021/ac2010197
[16]	SONG H P, ZHANG H, FU Y, et al. Screening for selective inhibitors of xanthine oxidase from Flos Chrysanthemum using ultrafiltration LC–MS combined with enzyme channel blocking[J]. Journal of Chromatography B,2014,961:56−61. doi: 10.1016/j.jchromb.2014.05.001
[17]	LI Y, DENG B, YANG S, et al. A colorimetric fluorescent probe for the detection of tyrosinase and its application for the food industry[J]. Journal of Photochemistry and Photobiology A Chemistry,2021,419(13):113458.
[18]	VALVERDE P, GARCIA J C, JIMENEZ-CERVANTES C, et al. Tyrosinase isoenzymes in mammalian melanocytes. 2. Differential activation by alpha-melanocyte-stimulating hormone[J]. Febs Journal,2010,217(2):541−548.
[19]	LIU M C, YANG S J, DA H, et al. A simple and convenient method for the preparation of antioxidant peptides from walnut (Juglans regia L. ) protein hydrolysates[J]. Chemistry Central Journal,2016,10:39−39. doi: 10.1186/s13065-016-0184-x
[20]	王计瑞, 谭均, 李隆云, 等. 国槐不同组织器官中多成分分析及抗氧化活性研究[J]. 中草药,2020,51(17):4513−4520. [WANG Jirui, TAN Jun, LI Longyun, et al. Multi-component analysis and antioxidant activity in different tissues and organs of Chinese Sophora japonicus[J]. Chinese Herbal Medicine,2020,51(17):4513−4520.
[21]	HIMANI Singh, MADHURI Kaushish Lily, KOUSHALYA Dangwal. Viburnum mullaha D. DON fruit (Indian Cranberry): A potential source of polyphenol with rich antioxidant, anti-elastase, anti-collagenase and anti-tyrosinase activities[J]. International Journal of Food Properties,2016,20(5-8):1729−1739.
[22]	王苗苗, 严欢, 田合, 等. 薰衣草提取物的体外抗氧化活性及酶抑制活性研究[J]. 食品工业科技,2019,40(16):14−26. [WANG Miaomiao, YAN Huan, TIAN He, et al. Study on the antioxidant activity and enzyme inhibition activity of lavender extract in vitro[J]. Food Industry Science and Technology,2019,40(16):14−26.
[23]	王丹, 时志春, 李军, 等. HPLC-ABTS⁺·在线检测快速评价茶叶总抗氧化能力[J]. 食品工业,2020,41(8):280−283. [WANG Dan, SHI Zhichun, LI Jun, et al. HPLC-ABTS⁺· on-line detection to quickly evaluate the total antioxidant capacity of tea[J]. Food Industry,2020,41(8):280−283.
[24]	陈菁, 周建桥, 谭国英, 等. 在线提取HPLC-ABTS联用快速分析枳椇子中抗氧化成分[J]. 药物分析杂志,2021,41(2):203−209. [CHEN Jing, ZHOU Jianqiao, TAN Guoying, et al. On-line extraction of HPLC-ABTS combined with rapid analysis of the antioxidant components in Hovenia dulcis[J]. Chinese Journal of Pharmaceutic,2021,41(2):203−209.
[25]	LIM, SOON, SUNG. Identification of tyrosinase specific inhibitors from Xanthium strumarium fruit extract using ultrafiltration-high performance liquid chromatography[J]. Journal of Chromatography B,2015,1002:319−328. doi: 10.1016/j.jchromb.2015.08.030
[26]	CHANG J L, PARK S K, JIN Y K, et al. Melanogenesis regulatory activity of the ethyl acetate fraction from Arctium lappa L. leaf on α-MSH–induced B16/F10 melanoma cells[J]. Industrial Crops and Products,2019,138:111581. doi: 10.1016/j.indcrop.2019.111581
[27]	王春宇, 姜民, 侯冠宇, 等. 小柴胡汤对过氧化氢诱导的人皮肤成纤维细胞氧化应激损伤的保护作用[J]. 中国实验诊断学,2018,22(6):1074−1077. [WANG Chunyu, JIANG Min, HOU Guanyu, et al. Protective effect of Xiaochaihu decoction on oxidative stress damage of human skin fibroblasts induced by hydrogen peroxide[J]. Chinese Laboratory Diagnosis,2018,22(6):1074−1077.
[28]	黄玲艳. 桂花提取物的抗氧化及延缓衰老作用研究[D]. 扬州: 扬州大学, 2017 WANG Lingyan. The antioxidant and anti-aging effects of Osmanthus fragrans extract[D]. Yangzhou: Yangzhou university, 2017.
[29]	WU L C, CHANG L H, CHEN S H, et al. Antioxidant activity and melanogenesis inhibitory effect of the acetonic extract of Osmanthus fragrans: A potential natural and functional food flavor additive[J]. LWT-Food Science and Technology,2009,42(9):1513−1519. doi: 10.1016/j.lwt.2009.04.004
[30]	FEI Z, PENG J, ZHAO Y. Varietal classification and antioxidant activity prediction of Osmanthus fragrans Lour. flowers using UPLC–PDA/QTOF–MS and multivariable analysis[J]. Food Chemistry,2017,217:490−497. doi: 10.1016/j.foodchem.2016.08.125
[31]	LIAO X, HONG Y, CHEN Z. Identification and quantification of the bioactive components in Osmanthus fragrans roots by HPLC-MS/MS[J]. Journal of Pharmaceutical Analysis,2021,11(3):299−307. doi: 10.1016/j.jpha.2020.06.010
[32]	钟询龙, 王若伦, 段炼, 等. 基于UPLC/Q-TOF-MS技术分析女贞子与墨旱莲配伍协同增效的物质基础[J]. 中国医院药学杂志,2017,37(19):1887−1891. [ZHONG Xunlong, WANG Ruolun, DUAN Lian, et al. Based on UPLC/Q-TOF-MS technology to analyze the material basis of the synergy of the compatibility of Ligustrum lucidum and Eclipta prostrata[J]. Chinese Journal of Hospital Pharmacy,2017,37(19):1887−1891.
[33]	夏玉英, 陈铃, 刘方乐, 等. LC-MS/MS法测定脑透析液中3种苯乙醇苷类成分的浓度及其体内外探针回收率考察[J]. 天然产物研究与开发,2017,29(11):1841−1845. [XIA Yuying, CHEN Ling, LIU Fangle, et al. Determination of the concentration of three phenethyl alcohol glycosides in cerebral dialysis fluid by LC-MS/MS and investigation of the probe recovery rate in vivo and in vitro[J]. Research and Development of Natural Products,2017,29(11):1841−1845.
[34]	尹伟, 宋祖荣, 刘金旗, 等. 桂花的化学成分研究[J]. 中国中药杂志,2015,40(4):679−685. [YI Wei, SONG Zurong, LIU Jinqi, et al. Study on the chemical constituents of Osmanthus fragrans[J]. Chinese Journal of Traditional Chinese Medicine,2015,40(4):679−685.
[35]	吴超然, 房仙颖, 萧伟. 桂花非挥发性成分及药理活性研究进展[J]. 天然产物研究与开发,2017,29(8):1439−1448. [WU Chaoran, FANG Xianyin, XIAO Wei. Research progress on non-volatile constituents and pharmacological activities of Osmanthus fragrans[J]. Natural Product Research and Development,2017,29(8):1439−1448.
[36]	林秀莲, 宋粉云, 潘玄玄, 等. UPLC-Q-TOF-MS分析补肾强身片中化学成分[J]. 中国实验方剂学杂志,2018,24(19):61−67. [LIN Xiulian, SONG Fenyun, PAN Xuanxuan, et al. Chemical constituents in the kidney tonifying and body strengthening tablets[J]. Chinese Journal of Experimental Formulas,2018,24(19):61−67.
[37]	CUI Y, WANG Q, SHI X, et al. Simultaneous quantification of 14 bioactive constituents in Forsythia suspensa by liquid chromatography-electrospray ionisation-mass spectrometry[J]. Phytochemical Analysis Pca,2010,21(3):253−260. doi: 10.1002/pca.1194
[38]	HAN J, MIN Y, HUI G, et al. Analysis of multiple constituents in a Chinese herbal preparation Shuang-Huang-Lian oral liquid by HPLC-DAD-ESI-MSn[J]. J Pharm Biomed Anal,2007,44(2):430−438. doi: 10.1016/j.jpba.2007.02.023
[39]	常冰. 不同桂花品种GC/MS和HPLC指纹图谱分析[D]. 开封: 河南大学, 2019 CHANG Bing. GC/MS and HPLC fingerprint analysis of different Osmanthus fragrans varieties[D]. Kaifeng: Henan University, 2019.
[40]	宋晓芳. 桂花中多酚类成分分析和过氧化麦角甾醇抗肿瘤活性研究[D]. 咸宁: 湖北科技学院, 2019 SONG Xiaofang. Analysis of polyphenols in Osmanthus fragrans and study on anti-tumor activity of ergosterol peroxide[D]. Xianning: Hubei University of Science and Technology, 2019.
[41]	吴金凤, 尹权微, 潘有智, 等. UPLC-Q-TOF-MS/MS快速分析苁蓉总苷胶囊化学成分[J]. 中国中药杂志,2016,41(17):3244−3251. [WU Jingfeng, YI Quanwei, PAN Youzhi, et al. UPLC-Q-TOF-MS/MS quick analysis of chemical components of cistanche glycoside capsules[J]. Chinese Journal of Traditional Chinese Medicine,2016,41(17):3244−3251.
[42]	梁世瓢, 刘飞, 杜飞飞, 等. LC-MS/MS法测定血浆中山奈酚-3-O-芸香糖苷浓度及其在大鼠药动学研究中的应用[J]. 中国临床药理学与治疗学,2006,11(5):491−496. [LIANG Shipiao, LIU Fei, DU Feifei, et al. LC-MS/MS method for determination of kaempferol-3-O-rutinoside in plasma and its application in rat pharmacokinetics[J]. Chinese Clinical Pharmacology and Therapeutics,2006,11(5):491−496.
[43]	赵明慧, 姜子涛, 刘韬, 等. 平榛叶中黄酮类化合物成分的研究[J]. 现代食品科技,2014,30(12):235−240. [ZHAO Minghui, JIANG Zitao, LIU Tao, et al. Study on the flavonoids in the leaves of Corylus heterophylla Fisch[J]. Modern Food Technology,2014,30(12):235−240.
[44]	元新霞, 桂花花期及采后储藏过程中连翘脂素含量变化规律研究[D]. 武汉: 华中科技大学, 2013 YUAN Xinxia, Study on the change of forsythiaside content in Osmanthus fragrans during flowering and postharvest storage[D]. Wuhan: Huazhong University of Science and Technology, 2013.
[45]	代纯, 靳熙茜, 汪海波, 等. 桂花乙醇萃取物的抗氧化活性及主成分结构的鉴定[J]. 武汉轻工大学学报,2019,38(1):9−16. [DAI Chun, JIN Xiqian, WANG Haibo, et al. Antioxidant activity of ethanol extract of Osmanthus fragrans and identification of main component structure[J]. Journal of Wuhan University of Light Industry,2019,38(1):9−16.
[46]	刘东茹, 任亚梅, 伯继芳, 等. 根皮苷及苹果幼果多酚对酪氨酸酶活性的抑制作用[J]. 现代食品科技,2019,35(8):113−119. [LIU Dongru, REN Yamei, BO Jifang, et al. Inhibition of phlorizin and polyphenols from green apple on tyrosinase activity[J]. Modern Food Technology,2019,35(8):113−119.
[47]	史莎莎, 阎胜利, 张方华. 维生素E对H₂O₂诱导的人甲状腺细胞凋亡的保护作用[J]. 免疫学杂志,2011,27(6):45−52. [SHI Shasha, YAN Shengli, ZHANG Fanghua. The protective effect of vitamin E on the apoptosis of human thyroid cells induced by H₂O₂[J]. Journal of Immunology,2011,27(6):45−52.
[48]	金明, 王玉娇, 金梅花, 等. 两种细胞建立肝细胞氧化损伤模型比较[J]. 中国公共卫生,2015,31(3):324−326. [JIN ming, WANG yujiao, JIN meihua, et al. Comparison of two kinds of cells to establish hepatocyte oxidative damage model[J]. Chinese Public Health,2015,31(3):324−326.
[49]	刘雷山. 两种皮肤细胞对过氧化氢的不同反应及CD147在氧化应激所致人皮肤成纤维细胞衰老中的机制研究[D]. 长沙: 中南大学, 2012 LIU Leishan. Study on the different reactions of two kinds of skin cells to hydrogen peroxide and the mechanism of CD147 in the senescence of human skin fibroblasts induced by oxidative stress[D]. Changsha: Zhongnan University, 2012.
[50]	WANG Z. Osmanthus fragrans flower aqueous extract and its enriched acteoside inhibit melanogenesis and ultraviolet-induced pigmentation[J]. Natural Product Communications,2018,13(5):575−580.

Supplements (0)

Cited By

Cited by

Periodical cited type(10)

1.	王睿敏，郑冬艳，但霞，李仁芳，曾庆坤，吴凤娇，黄丽，李玲. 具有潜在降压功能益生乳酸菌的筛选及其特性的研究. 食品科技. 2025(01): 1-8 .
2.	马新淼，魏敏敏，张左利，张轶腾，牛希跃，李雨鑫，李婕，许倩. 新疆哈萨克酸马奶中功能性乳酸菌株的筛选、鉴定及功能评价. 食品安全质量检测学报. 2024(07): 151-159 .
3.	刘怡雯，达久阿达，张敏，任秀梅，蒋绍平，田维，吴建平. 牦牛酸乳中乳酸菌的研究进展. 乳品与人类. 2024(04): 37-41 .
4.	雷善钰，江华明，李艳，梁锦鹏，张小平，赵珂，向泉桔，辜运富. 川西高原传统发酵牦牛乳奶酪中乳酸菌多样性及优良乳酸菌的筛选. 应用与环境生物学报. 2023(01): 27-34 .
5.	夏亚男，冯晨晨，韩荣，双全，额尔敦巴雅尔. 高产γ-氨基丁酸乳酸菌的筛选、鉴定及其益生特性研究. 食品科技. 2023(02): 14-20 .
6.	葛善赢，张海涛，王士佳，李佳宸，吴学智，张佰清. 脉冲强光诱变选育高产乳酸植物乳杆菌及其益生特性研究. 中国酿造. 2023(10): 59-64 .
7.	陈显玲，莫小群，杨琴，曾婷，苏龙. 植物乳杆菌XL-02发酵产γ-氨基丁酸条件的优化. 山东化工. 2022(06): 10-14 .
8.	马莉，刘慧燕，方海田，辛世华，李一鸣，贺捷群. 产γ-氨基丁酸乳酸菌的分离鉴定及其发酵条件优化. 中国酿造. 2022(07): 94-100 .
9.	莫小群，王雅，陈显玲，农秀丽，卢丽婷，杨福川，苏龙. 富含γ-氨基丁酸非乳益生菌香蕉发酵饮料工艺研究. 中国果菜. 2022(11): 20-26+31 .
10.	王玲芝，白雪，蒋咏梅. 正交试验法优化灵芝菌丝体γ-氨基丁酸提取工艺. 福建农业科技. 2022(10): 44-48 .