低共熔溶剂-球磨法提取竹叶黄酮碳苷工艺优化

doi:10.3969/j.issn.0253-2417.2023.04.009

摘要/Abstract

摘要：

以低共熔溶剂为提取溶剂，用球磨法从毛竹叶中提取黄酮碳苷类化合物，采用高效液相色谱-飞行时间质谱(LC-Q-TOF-MS)测定竹叶中4种黄酮碳苷(异荭草苷、荭草苷、异牡荆苷和牡荆苷)的含量，通过单因素试验和正交试验，对低共熔溶剂的含水量、提取时间、液料比等因素进行考察。结果表明，从竹叶中提取黄酮碳苷的最佳工艺为：低共熔溶剂(氯化胆碱和乙二醇构成)含水量50%、提取时间150 s、液料比6∶1(mL∶g)。在该条件下，从竹叶中提取4种黄酮碳苷的总提取得率为1.55 mg/g，其中，异荭草苷0.99 mg/g、荭草苷0.23 mg/g、牡荆苷0.04 mg/g、异牡荆苷0.29 mg/g。该方法提取得率比浸渍提取法提高了7.6%，且节约了时间和有机溶剂。利用该优化工艺，结合LC-Q-TOF-MS检测，成功应用于以毛竹叶为原料制成竹茶样品的黄酮碳苷提取与测定。

关键词: 黄酮碳苷, 球磨法, 低共熔溶剂, 提取, 竹叶

Abstract:

Ball mill method was used to extract flavone C-glycosides from bamboo leaves using deep eutectic solvents(DESs) as extraction solvent. The contents of four flavone C-glycosides(isoorientin, orientin, isovitexin and vitexin) in bamboo leaves were determined by liquid chromatography quadrupole time-of-flight mass spectrometry(LC-Q-TOF-MS). By single factor experiments and orthogonal design, the parameters including water content, extraction time and liquid-solid ratio of DESs were investigated. The results showed that the optimized extraction process of flavone C-glycosides from bamboo leaves was as follows: DESs(composed of choline chloride and ethylene glycol) water content of 50%, extraction time of 150 s, and liquid-solid ratio of 6∶1(mL∶g). Under these conditions, the total extraction yield of four flavone C-glycosides from bamboo leaves was 1.55 mg/g, including isoorientin of 0.99 mg/g, orientin of 0.23 mg/g, vitexin of 0.04 mg/g and isovitexin of 0.29 mg/g. The extraction yield of this method was 7.6% higher than that of immersion extraction, and the time and organic solvent were saved. This optimized technology combined with LC-Q-TOF-MS detection was successfully applied to the extraction and determination of flavone C-glycosides in bamboo tea samples made from bamboo leaves.

Key words: flavone C-glycosides, ball mill, deep eutectic solvents, extraction, bamboo leaves

中图分类号:

TQ35
R284.2

刘亚倩, 王进, 汤锋. 低共熔溶剂-球磨法提取竹叶黄酮碳苷工艺优化[J]. 林产化学与工业, 2023, 43(4): 60-66.

Yaqian LIU, Jin WANG, Feng TANG. Optimization of Extraction Process of Flavone C-glycosides from Bamboo Leaves by Deep Eutectic Solvent-Ball Mill Method[J]. Chemistry and Industry of Forest Products, 2023, 43(4): 60-66.

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参考文献 18

1	HU Y, ZHAO G, YIN F, et al. Effects of roasting temperature and time on aldehyde formation derived from lipid oxidation in scallop(Patinopecten yessoensis) and the deterrent effect by antioxidants of bamboo leaves[J/OL]. Food Chemistry, 2022, 369: 130936[2022-04-10]. https://doi.org/10.1016/j.foodchem.2021.130936.
2	YU Y , LI Z , CAO G , et al. Bamboo leaf flavonoids extracts alleviate oxidative stress in HepG2 cells via naturally modulating reactive oxygen species production and Nrf2-mediated antioxidant defense responses[J]. Journal of Food Science, 2019, 84 (6): 1609- 1620. doi: 10.1111/1750-3841.14609
3	ARAUJO SOUSA B, NASCIMENTO SILVA O, FARIAS PORTO W, et al. Identification of the active principle conferring anti-inflammatory and antinociceptive properties in bamboo plant[J/OL]. Molecules, 2021, 26(10): 3054[2022-04-10]. https://doi.org/10.3390/molecules26103054.
4	LIU L , PANG M , ZHANG Y . Lipase-catalyzed regioselective synthesis of flavone C-glucosides esters and high-efficiency oil-soluble antioxidant of bamboo leaves[J]. European Journal of Lipid Science and Technology, 2015, 117 (10): 1636- 1646. doi: 10.1002/ejlt.201400541
5	邹小琳, 吕兆林, 王媛媛, 等. 竹叶黄酮不同热加工方式稳定性[J]. 北京林业大学学报, 2016, 38 (11): 111- 117.
	ZOU X L , LYU Z L , WANG Y Y , et al. Stability analysis of bamboo leaf flavonoids under different thermal processing treatments[J]. Journal of Beijing Forestry University, 2016, 38 (11): 111- 117.
6	LI H , WANG X , LIU C , et al. An efficient pretreatment for the selectively hydrothermal conversion of corncob into furfural: The combined mixed ball milling and ultrasonic pretreatments[J]. Industrial Crops and Products, 2016, 94, 721- 728. doi: 10.1016/j.indcrop.2016.09.052
7	XIE J , SHI L , ZHU X , et al. Mechanochemical-assisted efficient extraction of rutin from Hibiscus mutabilis L[J]. Innovative Food Science & Emerging Technologies, 2011, 12 (2): 146- 152.
8	李大刚, 徐耀宗, 王志文, 等. CCD法探析三种辅助提取茶多酚工艺研究[J]. 茶叶通讯, 2021, 48 (4): 712- 716. doi: 10.3969/j.issn.1009-525X.2021.04.019
	LI D G , XU Y Z , WANG Z W , et al. Study on three assisted extraction processes of tea polyphenols by CCD experiment design[J]. Journal of Tea Communication, 2021, 48 (4): 712- 716. doi: 10.3969/j.issn.1009-525X.2021.04.019
9	贾永忠. 类离子液体[M]. 北京: 化学工业出版社, 2015.
	JIA Y Z . Ionic Liquid Analogues[M]. Beijing: Chemical Industry Press, 2015.
10	POTKA-WASYLKA J, RUTKOWSKA M, GUARDIA MDL. Are deep eutectic solvents useful inchromatography? A short review[J/OL]. Journal of Chromatography A, 2021, 1639: 461918[2022-04-10]. https://doi.org/10.1016/j.chroma.2021.461918.
11	岳少云, 金鑫, 董新月, 等. 低共熔溶剂提取香椿籽总黄酮效率的研究[J]. 阜阳师范学院学报(自然科学版), 2020, 37 (2): 67- 71.
	YUE S Y , JIN X , DONG X Y , et al. Extraction rate of total flavonoids from Toona sinensis seeds with different deep eutectic solvents[J]. Journal of Fuyang Normal University(Natural Science), 2020, 37 (2): 67- 71.
12	孔方, 李莉, 刘言娟. 超声辅助低共熔溶剂提取苹果叶中的总黄酮[J]. 食品工业科技, 2020, 41 (14): 134- 147.
	KONG F , LI L , LIU Y J . Ultrasonic-assisted deep eutectic solvents extraction of total flavonoids from apple leaves[J]. Science and Technology of Food Industry, 2020, 41 (14): 134- 147.
13	WANG J , ZHOU Y , WANG M , et al. High-throughput analysis for artemisinins with deep eutectic solvents mechanochemical extraction and direct analysis in real time mass spectrometry[J]. Analytical Chemistry, 2018, 90 (5): 3109- 3117. doi: 10.1021/acs.analchem.7b04060
14	WANG M , WANG J , ZHOU Y , et al. Ecofriendly mechanochemical extraction of bioactive compounds from plants with deep eutectic solvents[J]. ACS Sustainable Chemistry & Engineering, 2017, 5 (7): 6297- 6303.
15	WANG M , WANG J , ZHANG Y , et al. Fast environment-friendly ball mill-assisted deep eutectic solvent-based extraction of natural products[J]. Journal of Chromatography A, 2016, 1443, 262- 266.
16	WANG J, YUE Y, JIANG H, et al. Rapid screening for flavone C-glycosides in the leaves of different species of bamboo and simultaneous quantitation of four marker compounds by HPLC-UV/DAD[J/OL]. International Journal of Analytical Chemistry, 2012, 2012: 205101[2022-04-10]. https://10.1155/2012/20510.
17	檀珮雯. 毛竹有效成分提取的关键技术及其手工皂的研究[D]. 合肥: 安徽农业大学, 2019.
	TAN P W. Key techniques of bamboo active components extracting and its research on handmand soap[D]. Hefei: Anhui Agricultrual University, 2019.
18	娄文勇. 离子液体中生物催化不对称反应研究[M]. 广州: 华南理工大学出版社, 2017.
	LOU W Y . Study on the Biocatalytic Asymmetric Reactions in Ionic Liquids[M]. Guangzhou: South China University of Technology Press, 2017.

低共熔溶剂 deep eutectic solvents(DESs)	氢键受体 hydrogen bond acceptor(HBA)	氢键供体 hydrogen bond donor(HBD)	氢键受体/氢键供体物质的量比 molar ratio of HBA/HBD
DES-1	氯化胆碱choline chloride	乙二醇ethylene glycol	1∶2
DES-2	氯化胆碱choline chloride	1, 4-丁二醇1, 4-butane-diol	1∶4
DES-3	氯化胆碱choline chloride	尿素carbamide	1∶2
DES-4	氯化胆碱choline chloride	柠檬酸citric acid	1∶1

序号 No.	保留时间/min retention time	准分子离子峰 [M+H]⁺	误差 error	精确相对分子质量 accurate relative molecular mass	分子式 molecular formula	鉴定 identification
1	5.34	449.107 0	2.25	448.100 6	C₂₁H₂₀O₁₁	异荭草苷isoorientin
2	6.19	449.106 7	3.44	448.100 6	C₂₁H₂₀O₁₁	荭草苷orientin
3	9.03	433.112 0	2.97	432.105 6	C₂₁H₂₀O₁₀	牡荆苷vitexin
4	9.39	433.112 4	1.83	432.105 6	C₂₁H₂₀O₁₀	异牡荆苷isovitexin

编号 No.	A DES-1含水量/% water content of DES-1	B 提取时间/s extraction time	C 液料比(mL∶g) liquid-to-solid ratio	4种黄酮碳苷总得率/(mg·g^-1) total yield of four flavone C-glycosides
1	40	120	6∶1	1.51
2	40	150	7∶1	1.52
3	40	180	8∶1	1.13
4	50	120	7∶1	1.49
5	50	150	8∶1	1.51
6	50	180	6∶1	1.55
7	60	120	8∶1	1.49
8	60	150	6∶1	1.52
9	60	180	7∶1	1.52
k₁	1.39	1.50	1.53
k₂	1.52	1.52	1.51
k₃	1.51	1.40	1.38
R	0.13	0.12	0.15

方差来源 source	离差平方和 sum of squares	自由度 df	均方 mean square	F值 F value	显著性 significant
A	0.032	2	0.016	0.859	p>0.05
B	0.023	2	0.012	0.642	p>0.05
C	0.041	2	0.020	1.084	p>0.05
F_{0.05(2, 2)}=19.00

提取方法 extraction method	提取溶剂 extraction solvent	提取时间 extraction time	4种黄酮碳苷总得率¹⁾/(mg·g^-1) total yield of four flavone C-glycosides	文献 ref.
低共熔溶剂-球磨法 ball mill assisted deep eutectic solvent extraction	n(氯化胆碱)/n(乙二醇)(1∶2) n(choline chloride)/n(ethylene glycol)	2.5 min	1.55±0.03 a	本研究 this study
超声波提取 ultrasonic extraction	V(甲醇)/V(水)(6∶4) V(methanol)/V(water)	90 min	1.60±0.04 a	[16]
浸渍提取 soaking method extraction	V(乙醇)/V(水)(8∶2) V(ethanol)/V(water)	48 h	1.44±0.01 b	[17]