Extraction Process and Antioxidant Activities Screening of Total Flavonoids from Citrus depressa Hayat Leaves

doi:10.11924/j.issn.1000-6850.casb2024-0534

Abstract

Abstract:

This study aimed to optimize the extraction process of total flavonoids from Citrus depressa Hayat leaves (CDHL), and investigate their antioxidant activities. Total flavonoids yield was used as evaluation index, the effects of extraction solvent, solid-liquid ratio, extraction time, and extraction frequency on the total flavonoids yield from CDHL were investigated by single-factor experiments. The extraction condition was optimized by orthogonal experiment. The antioxidant activity of total flavonoids was studied by measuring the removal capacity of DPPH, ABTS, hydroxyl radicals and the reducing capacity were investigated in vitro. The best extraction process conditions for total flavonoids from CDHL were distilled water reflux for 1 h and 2 times, with liquid ratio of 1:25 (g/mL). The mean extraction of total flavonoids under this condition was (32.60±2.16) mg/g. The IC₅₀ of DPPH, ABTS and hydroxyl radicals were 12.03, 105.79 and 1.10 μg/mL respectively, and its reducing ability was strong. This extraction process was stable and reliable for extracting total flavonoids from CDHL. The total flavonoids from CDHL can be used as a novel natural antioxidant in functional foods.

Key words: Citrus depressa Hayat leaves, total flavonoids, extraction technology, orthogonal test, antioxidation

WANG Jie, YU Xiaoxia, HE Jianzhang, CHEN Jiantu, WU Meifang. Extraction Process and Antioxidant Activities Screening of Total Flavonoids from Citrus depressa Hayat Leaves[J]. Chinese Agricultural Science Bulletin, 2025, 41(8): 133-139.

Figures/Tables 11

References 27

[1]	赖瑞云, 黄珺梅, 林建忠, 等. 台湾香檬在厦门地区的引种初报[J]. 亚热带植物科学, 2018, 47(3):289-291.
[2]	SALMAH Y, MOHD G H, SWEE K G. Naringin content in local citrus fruits[J]. Food chemistry, 1990, 37(2):113-121.
[3]	谷乐, 刘韶松, 何丽娟, 等. 台湾香檬果皮川陈皮素分离制备及其抗肿瘤活性研究[J]. 亚热带植物科学, 2021, 50(2):83-87.
[4]	CHIEN W, SAPUTRI D S, LIN H. Valorization of Taiwan's Citrus depressa Hayata peels as a source of nobiletin and tangeretin using simple ultrasonic-assisted extraction[J]. Current research in food science, 2022, 5:278-287.
[5]	YONATHAN A, GOKI M, HAJIME T, et al. Cultivation line and fruit ripening discriminations of Shiikuwasha (Citrus depressa Hayata) peel oils using aroma compositional, electronic nose, and antioxidant analyses[J]. Food research international, 2015, 67(Jan.): 108-110.
[6]	MINAGAWA A, OTANI Y, KUBOTA T, et al. The citrus flavonoid, nobiletin, inhibits peritoneal dissemination of human gastric carcinoma in SCID mice[J]. Japanese journal of cancer research: Gann, 2001, 92(12):1322-1328.
[7]	SHINICHI U, HIROFUMI Y, RYU M, et al. Synergistic antitumor effect of a combination of paclitaxel and carboplatin with nobiletin from Citrus depressa on non-small-cell lung cancer cell lines[J]. Planta medica, 2014, 80(6):452-457. doi: 10.1055/s-0034-1368321 pmid: 24687742
[8]	AKACHI T, SHIINA Y, OHISHI Y, et al. Hepatoprotective effects of flavonoids from Shekwasha (Citrus depressa) against D-galactosamine-induced liver injury in rats[J]. Journal of nutritional science and vitaminology, 2010, 56(1):60-67. pmid: 20354348
[9]	EUN Y L, SE H K, SUKKUM N C, et al. Efficacy of polymethoxylated flavonoids from citrus depressa extract on alcohol-induced liver injury in mice[J]. Biotechnology and bioprocess engineering, 2019, 24(6):907-914.
[10]	CHIEN W, SAPUTRI D S, LIN H Y. Valorization of Taiwan's Citrus depressa Hayata peels as a source of nobiletin and tangeretin using simple ultrasonic-assisted extraction[J]. Current research in food science, 2022,5:278-287.
[11]	YONATHAN A, IKUKO T, SAYURI I, et al. Volatile aroma components and antioxidant activities of the flavedo peel extract of unripe Shiikuwasha (Citrus depressa Hayata)[J]. Journal of food science, 2012, 77(4):445-469.
[12]	KAZUNA M, SHOKO W, TOMOKO F, et al. Quality improvement of Shiikuwasha (Citrus depressa Hayata) juice by combining anion exchange treatment and insoluble pulp addition[J]. Journal of the Japanese society for food science and technology-nippon Shokuhin Kagaku Kogaku Kaishi, 2009, 56(4):193-199.
[13]	KAZUNA M, TOMOKO F, NOBUYUKI K, et al. Effect of some extracting methods and storage temperature on quality stability of Shiikuwasha (Citrus depressa Hayata) fruit juice during storage: stability of polymethoxylated flavones and synephrine[J]. Food preservation science, 2009, 35(1):3-9.
[14]	王杰. 驯化台湾香檬中川陈皮素与辛弗林的鉴别与测定[J]. 海峡药学, 2022, 34(12):48-52.
[15]	孙国艳, 李华, 杨长花, 等. 响应面法优化淫羊藿总黄酮的提取工艺[J]. 化学与生物工程, 2022, 39(11):37-41.
[16]	刘威良, 蔡建, 高文荣, 等. 云南野生火棘叶甾醇提取工艺优化及体外活性研究[J]. 食品工业科技, 2024, 45(11):1-8.
[17]	郭瑛, 赵文惠, 李安林, 等. 疏花蔷薇花总多酚提取工艺优化及其抗氧化活性[J]. 食品研究与开发, 2024, 45(3):116-122.
[18]	王源, 宋梦楠, 刘忍阳, 等. 杜鹃兰叶总黄酮提取工艺的优化及抗氧化活性研究[J]. 湖北医药学院学报, 2023, 42(1):1-6.
[19]	陈桐, 侯林清, 覃凡倚, 等. 竹叶柴胡总黄酮提取及抗氧化作用研究[J]. 辽宁化工, 2023, 52(2):187-189.
[20]	夏雨弘, 刘颖, 周茗, 等. 小二仙草总黄酮提取工艺的优化及其抗氧化活性分析[J]. 食品工业科技, 2023, 44(18):244-250.
[21]	李东香, 关荣发, 黄海智, 等. 3种新疆沙棘黄酮的提取优化及抗氧化活性对比[J]. 中国食品学报, 2023, 23(4):157-167.
[22]	桂利利, 吴正坤, 余惠凡, 等. 黄药叶总黄酮提取工艺优化及其抗氧化活性研究[J]. 食品研究与开发, 2021, 42(19):100-108.
[23]	李金金, 孟琦, 崔馨燕, 等. 响应面法优化地参三萜酸提取工艺及抗氧化活性分析[J]. 南方农业学报, 2022, 53(8):2261-2271.
[24]	李兰, 段政, 汤有宏, 等. 金银花总黄酮的提取工艺及其抗氧化性研究[J]. 酿酒, 2022, 49(6):131-136.
[25]	贺敬霞, 牟倩倩, 杨滨, 等. 中药黄酮类成分提取工艺研究进展[J]. 亚太传统医药, 2017, 13(15):30-36. doi: 10.11954/ytctyy.201715013
[26]	王慧. 低共熔溶剂在黄芩有效成分提取及制备中的应用研究[D]. 太原: 山西大学, 2019.
[27]	李婷婷. 微波辅助低共熔溶剂提取黄芩中主要黄酮成分研究[D]. 哈尔滨: 东北林业大学, 2014.

水平	A时间/h	B次数/次	C料液比/(g/mL)
1	0.5	1	1:15
2	1	2	1:20
3	1.5	3	1:25

水平	A时间/h	B次数/次	C料液比/(g/mL)
1	0.5	1	1:15
2	1	2	1:20
3	1.5	3	1:25

试验号	因素				提取率/ (mg/g)
试验号	A提取时间/h	B提取次数/次	C料液比/ (g/mL)	D空白	提取率/ (mg/g)
1	1	1	1	1	23.64
2	1	2	2	2	30.28
3	1	3	3	3	30.01
4	2	1	2	3	26.76
5	2	2	3	1	31.62
6	2	3	1	2	29.49
7	3	1	3	2	26.94
8	3	2	1	3	28.89
9	3	3	2	1	30.87
K₁	27.98	25.79	27.34	28.71	—
K₂	29.29	30.26	29.30	28.90	—
K₃	28.90	30.12	29.52	28.55	—
R	1.31	4.48	2.18	0.35	—

试验号	因素				提取率/ (mg/g)
试验号	A提取时间/h	B提取次数/次	C料液比/ (g/mL)	D空白	提取率/ (mg/g)
1	1	1	1	1	23.64
2	1	2	2	2	30.28
3	1	3	3	3	30.01
4	2	1	2	3	26.76
5	2	2	3	1	31.62
6	2	3	1	2	29.49
7	3	1	3	2	26.94
8	3	2	1	3	28.89
9	3	3	2	1	30.87
K₁	27.98	25.79	27.34	28.71	—
K₂	29.29	30.26	29.30	28.90	—
K₃	28.90	30.12	29.52	28.55	—
R	1.31	4.48	2.18	0.35	—

方差来源	离差平方和	自由度	均方	F	P
A提取时间	2.73	2	1.36	14.83	>0.05
B提取次数	38.98	2	19.48	211.87	<0.05
C料液比	8.67	2	4.34	47.12	<0.05
误差	0.18	2	0.09	—	—