芦丁在大鼠体内的代谢产物鉴定与代谢途径分析

doi:10.3976/j.issn.1002-4026.2023.05.002

山东科学 ›› 2023, Vol. 36 ›› Issue (5): 9-18.doi: 10.3976/j.issn.1002-4026.2023.05.002

芦丁在大鼠体内的代谢产物鉴定与代谢途径分析

王常瞵^a(), 高明周^b, 郭英慧^a, 孙亚^b, 于小钧^c, 闫志^c, 王杰琼^c^,^*(), 乔明琦^a^,^*()

a.山东中医药大学中医学院,山东济南 250355
b.山东中医药大学中医药创新研究院,山东济南 250355
c.山东中医药大学药学院,山东济南 250355

收稿日期:2022-10-19 出版日期:2023-10-20 发布日期:2023-10-12
通信作者: 王杰琼,乔明琦 E-mail:wcl19960125@outlook.com;jieqiong2016@126.com;qmingqi@sdutcm.com
作者简介:王常瞵(1996—),男,研究方向为肝藏象病理生理与情志致病机理研究。E-mail:wcl19960125@outlook.com
基金资助:
国家自然科学基金面上项目(81473558);山东省自然科学基金(ZR202102270167);国家科技重大专项(重大新药创制)(SQ2017ZX091064);山东省自然科学基金重点项目(ZR2020ZD17);山东省医药卫生科技发展计划(202105010467)

Identification of rutin metabolites and analysis of rutin metabolic pathway in rats

WANG Changlin^a(), GAO Mingzhou^b, GUO Yinghui^a, SUN Ya^b, YU Xiaojun^c, YAN Zhi^c, WANG Jieqiong^c^,^*(), QIAO Mingqi^a^,^*()

a. School of Traditional Chinese Medicine, Shandong University of Traditional Chinese Medicine, Jinan 250355, China
b. Institute of Traditional Chinese Medicine Innovation, Shandong University of Traditional Chinese Medicine, Jinan 250355, China
c. School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, China

Received:2022-10-19 Online:2023-10-20 Published:2023-10-12
Contact: WANG Jieqiong, QIAO Mingqi E-mail:wcl19960125@outlook.com;jieqiong2016@126.com;qmingqi@sdutcm.com

摘要/Abstract

摘要：

分析大鼠口服芦丁后血浆、尿液和粪便中的代谢产物并评价芦丁在大鼠体内的代谢途径。大鼠灌胃给予250 mg/kg芦丁后,采集血浆、尿液和粪便,以固相萃取法处理生物样品,利用超高效液相色谱-四级杆-静电场轨道阱质谱,以0.05%甲酸水(A)-0.05%甲酸乙腈(B)为流动相梯度洗脱,分别在正、负离子模式下采集各样品数据,通过高分辨提取离子流图和平行反应监测模式,结合色谱保留时间、质量测量、诊断离子等确定芦丁的代谢物并探究芦丁在大鼠体内的代谢途径。在正、负离子模式下共检测和鉴定了29种芦丁代谢产物,其主要代谢途径为甲基化、葡萄糖醛酸化、硫酸酯化及其复合反应等。建立了芦丁的整体代谢图谱,为其进一步的药效学评价及开发利用提供了参考依据,同时也为药物代谢鉴定研究提供了一种综合研究方法。

关键词: 芦丁, 超高效液相色谱-四级杆-静电场轨道阱质谱, 代谢产物, 代谢途径

Abstract:

Herein, metabolites in plasma, urine and feces of rats were analyzed after oral administration of rutin and the metabolic pathway of rutin was evaluated. After intragastric administration of 250 mg/kg rutin, plasma, urine, and feces were collected and treated via solid phase extraction. Ultra-high performance liquid chromatography-Q Exactive hybrid quadrupole-orbitrapmass spectrometry (UPLC-Q-Orbitrap MS) was used with 0.05% formic acid water (A)-0.05% formic acid acetonitrile (B) as mobile phase gradient elution. The sample data were collected in positive- and negative-ion modes. The metabolites and metabolic pathway of rutin in rats were determined via high resolution extraction ion chromatography in the parallel reaction monitoring mode, combined with chromatographic retention time, accurate mass measurement and diagnostic ions.Twenty-nine rutin metabolites were detected and identified in positive and negative ion modes,and their main metabolic pathways were methylation, glucuronidation, sulfation and their compound reaction. The study provided the overall metabolic profile of rutin, which will provide a reference for further pharmacodynamic evaluation, development, and utilization in the future and offer a comprehensive research method for drug metabolism identification.

Key words: rutin, UPLC-Q-Orbitrap MS, metabolites, metabolic pathways

中图分类号:

R285

王常瞵, 高明周, 郭英慧, 孙亚, 于小钧, 闫志, 王杰琼, 乔明琦. 芦丁在大鼠体内的代谢产物鉴定与代谢途径分析[J]. 山东科学, 2023, 36(5): 9-18.

WANG Changlin, GAO Mingzhou, GUO Yinghui, SUN Ya, YU Xiaojun, YAN Zhi, WANG Jieqiong, QIAO Mingqi. Identification of rutin metabolites and analysis of rutin metabolic pathway in rats[J]. Shandong Science, 2023, 36(5): 9-18.

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参考文献 23

[1]	马溶, 庞广昌. 芦丁对现代文明病的作用[J]. 食品科学, 2013, 34(7): 307-311. doi: 10.7506/spkx1002-6630-201307065
[2]	AL-DHABI N A, ARASU M V, PARK C H, et al. An up-to-date review of rutin and its biological and pharmacological activities[J]. EXCLI Journal, 2015, 14: 59-63. DOI: 10.17179/excli2014-663. pmid: 26535031
[3]	MAULUDIN R, MÜLLER R H, KECK C M. Development of an oral rutin nanocrystal formulation[J]. International Journal of Pharmaceutics, 2009, 370(1/2): 202-209. DOI: 10.1016/j.ijpharm.2008.11.029.
[4]	WEBSTER R P, GAWDE M D, BHATTACHARYA R K. Protective effect of rutin, a flavonol glycoside, on the carcinogen-induced DNA damage and repair enzymes in rats[J]. Cancer Letters, 1996, 109(1/2): 185-191. DOI: 10.1016/S0304-3835(96)04443-6.
[5]	NEGAHDARI R, BOHLOULI S, SHARIFI S, et al. Therapeutic benefits of rutin and its nanoformulations[J]. Phytotherapy Research, 2021, 35(4): 1719-1738. DOI: 10.1002/ptr.6904. pmid: 33058407
[6]	CARBONARO M, GRANT G. Absorption of quercetin and rutin in rat small intestine[J]. Annals of Nutrition & Metabolism, 2005, 49(3): 178-182. DOI: 10.1159/000086882.
[7]	YANG J, QIAN D W, JIANG S, et al. Identification of rutin deglycosylated metabolites produced by human intestinal bacteria using UPLC-Q-TOF/MS[J]. Journal of Chromatography B,AnalyticalTechnologiesin the Biomedical and Life Sciences, 2012, 898: 95-100. DOI: 10.1016/j.jchromb.2012.04.024.
[8]	周鹏飞, 王建壮, 庞小雄, 等. 离体培养人肠道菌群对芦丁代谢的研究[J]. 广东药学院学报, 2011, 27(6): 582-586. DOI: 10.3969/j.issn.1006-8783.2011.06.008.
[9]	伍明江, 吴晓磊, 张德芹, 等. UPLC-Q-TOF/MS鉴定芦丁在大鼠体内的代谢产物[J]. 中国实验方剂学杂志, 2017, 23(17): 91-97. DOI: 10.13422/j.cnki.syfjx.2017170091.
[10]	邓志鹏, 张喆, 周红燕, 等. 异绿原酸C大鼠体内代谢产物鉴定[J]. 世界科学技术-中医药现代化, 2022, 24(5): 2071-2078.
[11]	吴励萍, 张祎盈, 冯一帆, 等. 杞菊景明颗粒在大鼠血浆、胆汁、尿液、粪便中代谢产物的鉴定[J]. 中成药, 2022, 44(6): 1725-1731. DOI: 10.3969/j.issn.1001-1528.2022.06.001.
[12]	TAMÁS K, WACHTER-KISS E, KORMÁNY R. Hydrazine determination in allopurinol using derivatization and SPE for sample preparation[J]. Journal of Pharmaceutical and Biomedical Analysis, 2018, 152: 25-30. DOI: 10.1016/j.jpba.2018.01.035. pmid: 29414015
[13]	JAGADEESAN K K, ROSSETTI C, ABDELQADER A, et al. Filter plate-based screening of MIP SPE materials for capture of the biomarker pro-gastrin-releasing peptide[J]. SLAS Discovery: Advancing Life Sciences R & D, 2017, 22(10): 1253-1261. DOI: 10.1177/2472555216689494.
[14]	WANG F, CAO G S, LI Y, et al. Characterization of forsythoside A metabolites in rats by a combination of UHPLC-LTQ-Orbitrap mass spectrometer with multiple data processing techniques[J]. Biomedical Chromatography, 2018, 32(5): e4164. DOI: 10.1002/bmc.4164.
[15]	XIANG Z D, WANG S P, LI H R, et al. Detection and identification of catalpol metabolites in the rat plasma, urine and faeces using ultra-high performance liquid chromatography-Q exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry[J]. Current Drug Metabolism, 2021, 22(3): 173-184. DOI: 10.2174/1389200221999201125205515. pmid: 33243112
[16]	崔议方, 王红, 阮健, 等. 基于UHPLC-Q-Exactive Orbitrap MS鉴定欧前胡素在大鼠体内的代谢产物[J]. 中国中药杂志, 2022, 47(15): 4202-4213. DOI: 10.19540/j.cnki.cjcmm.20220307.203.
[17]	JIANG S, WANG S P, DONG P P, et al. A comprehensive profiling and identification of liquiritin metabolites in rats using ultra-high-performance liquid chromatography coupled with linear ion trap-orbitrap mass spectrometer[J]. Xenobiotica, 2021, 51(5): 564-581. DOI: 10.1080/00498254.2020.1854366. pmid: 33222601
[18]	DONG P P, SHI L, WANG S P, et al. Rapid profiling and identification of vitexin metabolites in rat urine, plasma and faeces after oral administration using a UHPLC-Q-exactive orbitrap mass spectrometer coupled with multiple data-mining methods[J]. Current Drug Metabolism, 2021, 22(3): 185-197. DOI: 10.2174/1389200221999210101232841. pmid: 33397253
[19]	LIU M H, TONG X, WANG J X, et al. Rapid separation and identification of multiple constituents in traditional Chinese medicine formula Shenqi Fuzheng Injection by ultra-fast liquid chromatography combined with quadrupole-time-of-flight mass spectrometry[J]. Journal of Pharmaceutical and Biomedical Analysis, 2013, 74: 141-155. DOI: 10.1016/j.jpba.2012.10.024.
[20]	YE J B, REN G, LI W Y, et al. Characterization and identification of prenylated flavonoids from Artocarpus heterophylluslam. roots by quadrupole time-of-flight and linear trap quadrupole orbitrap mass spectrometry[J]. Molecules (Basel, Switzerland), 2019, 24(24): 4591. DOI: 10.3390/molecules24244591.
[21]	LIU X C, FAN X Y, WANG X, et al. Structural characterization and screening of chemical markers of flavonoids in Lysimachiae Herba and Desmodii Styracifolii Herba by ultra high-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry based metabolomics approach[J]. Journal of Pharmaceutical and Biomedical Analysis, 2019, 171: 52-64. DOI: 10.1016/j.jpba.2019.04.003. pmid: 30965221
[22]	QU Y X, ZHANG C N, LIU R H, et al. Rapid characterization the chemical constituents of Bergenia purpurascens and explore potential mechanism in treating osteoarthritis by ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry combined with network pharmacology[J]. Journal of Separation Science, 2020, 43(16): 3333-3348. DOI: 10.1002/jssc.201901284.
[23]	叶馨源, 吴建明, 杨杰, 等. 白背三七化学成分及代表性成分质谱裂解特征的研究进展[J]. 中草药, 2021, 52(21):6687-6700. DOI: 10.7501/j.issn.0253-2670.2021.21.028.

序号	离子模式		t_R/min		推测化学式		理论分子量		实验分子量		误差		MS/MS碎片信息
M0	N		3.81		C₂₇H₂₉O₁₆		609.145 01		609.146 48		0.872×10^-6		609(100.00),301(29.76),151(4.87),179 (3.51),273(0.85),107(0.20)
M1	N		3.49		C₂₈H₂₇O₁₉		667.114 10		667.115 42		2.783×10^-6		315(100.00),667(71.99),491(51.74),301 (17.59),151(0.61),179(0.58)
M2	N		3.52		C₂₇H₂₉O₁₉S		689.101 82		689.103 58		1.015×10^-6		689(100.00),301(42.89),381(35.28),179 (3.96),151(3.07)
M3	N		3.79		C₂₁H₁₇O₁₆S		557.023 18		557.023 80		0.873×10^-6		301(100.00),557(50.79),477(16.46),151 (5.76),179(4.89)
M4	N		4.41		C₁₇H₁₃O₇		329.065 57		329.063 40		-4.650×10^-6		329(100.00),165(1.33),121(0.73)
M5	N		4.46		C₁₆H₁₁O₁₀S		395.006 74		395.007 54		2.016×10^-6		315(100.00),395(40.52),151(6.69),301 (5.17)
M6		N		4.69		C₁₈H₁₅O₇		343.081 22		343.082 85		3.468×10^-6		343(100.00),299(1.59),165(0.94),313 (0.28),121(0.15)
M7		N		6.09		C₂₉H₃₃O₁₆		637.176 31		637.177 16		0.428×10^-6		329(100.00),637(4.69),179(1.50),207 (0.50)

序号	可能的代谢物描述	尿液	血浆	粪便
M0	芦丁	+	+	+
M1	甲基槲皮素二葡萄糖醛酸	+	-	-
M2	*芦丁硫酸酯化产物	-	+	+
M3	*槲皮素葡萄糖醛酸的硫酸酯化产物	+	-	-
M4	*二甲基槲皮素	-	+	-
M5	甲基槲皮素硫酸酯	+	-	-
M6	*三甲基槲皮素	+	-	-
M7	*二甲基芦丁	+	+	-

芦丁在大鼠体内的代谢产物鉴定与代谢途径分析

Identification of rutin metabolites and analysis of rutin metabolic pathway in rats

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