滨海刺芹愈伤组织提取物中4种多酚含量测定及抗氧化、抗炎活性研究

doi:10.3969/j.issn.2097-2806.2024.01.008

摘要/Abstract

摘要：

采用热水回流法制备滨海刺芹愈伤组织提取物，基于高效液相色谱法建立同时测定滨海刺芹愈伤组织提取物中迷迭香酸、隐绿原酸、金丝桃苷、槲皮素-3-O-（6''-O-乙酰基）-β-D-吡喃葡萄糖苷四种多酚含量的方法，测定提取物对DPPH自由基、ABTS自由基清除能力。同时通过CCK-8法检测筛选样品给药浓度，建立痤疮丙酸杆菌诱导HaCaT细胞炎症模型，评价其抗炎功效。结果表明，提取物中隐绿原酸含量最高，为5.23 mg/g;迷迭香酸、槲皮素-3-O-（6''-O-乙酰基）-β-D-吡喃葡萄糖苷、金丝桃苷的含量依次为1.30，0.50，0.08 mg/g。提取物具有较好的抗氧化、抗炎能力，对DPPH自由基、ABTS自由基清除能力的IC₅₀值分别为0.323 9，0.296 5 mg/mL，对痤疮丙酸杆菌诱导的HaCaT细胞分泌IL-1β，IL-8也具有显著抑制作用（P<0.05）。

关键词: 滨海刺芹, 高效液相色谱, 多酚, 抗氧化, 抗炎

Abstract:

The preparation of extracts from callus of Eryngium maritimum was conducted by hot water reflux method. A method for simultaneous determination of rosmarinic acid, cryptochlorogenic acid, hyperoside, quercetin-3-O-（6''-O-acetyl）-β-D-glucopyranoside in the callus extract of Eryngium maritimum was established based on high performance liquid chromatography. The scavenging ability of the extracts on DPPH and ABTS free radicals were determined. The screening samples were also assayed by the CCK-8 method to determine the concentration of drug administration and a model of HaCaT cell inflammation induced by Cutibacterium acnes was established to evaluate its anti-inflammatory efficacy. The results show that the extracts contain the highest amount of cryptochlorogenic acid at 5.23 mg/g, and the contents of rosmarinic acid, quercetin-3-O-（6''-O-acetyl）-β-D-glucopyranoside and hyperin were 1.30, 0.50 and 0.08 mg/g, respectively. The extracts display antioxidant and anti-inflammatory abilities. The IC₅₀ values of the extracts on DPPH and ABTS free radical scavenging ability are 0.323 9 and 0.296 5 mg/mL, respectively, and also have significant inhibitory effects on the secretion of IL-1β and IL-8 in HaCaT cells induced by Cutibacterium acnes （P<0.05）.

Key words: Eryngium maritimum L., high performance liquid chromatography, polyphenol, anti-oxidant, anti-inflammatory

中图分类号:

TQ658

肖雪, 张慧荣, 李硕雯, 李孙华, 郭苗苗, 李丽. 滨海刺芹愈伤组织提取物中4种多酚含量测定及抗氧化、抗炎活性研究[J]. 日用化学工业（中英文）, 2024, 54(1): 57-64.

Xue Xiao, Huirong Zhang, Shuowen Li, Sunhua Li, Miaomiao Guo, Li Li. Determination of four polyphenols and study on antioxidant and anti-inflammatory activities of extracts from callus of Eryngium maritimum L.[J]. China Surfactant Detergent & Cosmetics, 2024, 54(1): 57-64.

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编号	种类	回归方程	相关系数（r）	线性范围/ （mg/L）
1	隐绿原酸	y=17.147x-66.697	0.999 8	9~450
2	槲皮素-3-O-（6''-O-乙酰基）-β-D-吡喃葡萄糖苷	y=27.292x+2.858 7	0.999 8	2~100
3	金丝桃苷	y=16.094x+1.085 5	0.999 3	1~ 50
4	迷迭香酸	y=24.637x+7.484 9	0.999 8	8~400

成分	峰面积/（mAU·s）					RSD/%
成分	混标-1	混标-2	混标-3	混标-4	混标-5	RSD/%
隐绿原酸	1 846.16	1 859.83	1 859.87	1 863.17	1 866.65	0.42
槲皮素-3-O-（6''-O-乙酰基）-β-D-吡喃葡萄糖苷	1 151.55	1 153.67	1 153.59	1 151.91	1 152.89	0.08
金丝桃苷	445.12	443.24	461.26	444.29	431.05	2.42
迷迭香酸	2 120.35	2 135.98	2 136.58	2 141.29	2 140.23	0.40

成分	含量/（mg/g）			平均含量/（mg/g）	RSD/%
成分	样品1	样品2	样品3	平均含量/（mg/g）	RSD/%
隐绿原酸	5.35	5.22	5.13	5.23	2.11
槲皮素-3-O-（6''-O-乙酰基）-β-D-吡喃葡萄糖苷	0.51	0.50	0.50	0.50	1.15
金丝桃苷	0.08	0.08	0.08	0.08	0
迷迭香酸	1.32	1.27	1.32	1.30	2.22

成分	峰面积/（mAU·s）				RSD/%
成分	0 h	6 h	12 h	24 h	RSD/%
隐绿原酸	1 836.16	1 829.83	1 863.17	1 866.65	1.01
槲皮素-3-O-（6''-O-乙酰基）-β-D-吡喃葡萄糖苷	1 154.55	1 153.67	1 159.91	1 152.89	0.27
金丝桃苷	445.12	433.24	444.29	441.05	1.23
迷迭香酸	2 120.35	2 135.98	2 131.29	2 140.23	0.40

成分	加标回收率/%			平均回收率/%	RSD/%
成分	样品1	样品2	样品3	平均回收率/%	RSD/%
隐绿原酸	98.21	101.54	100.35	100.03	1.69
槲皮素-3-O-（6''-O-乙酰基）-β-D-吡喃葡萄糖苷	99.25	98.74	101.27	99.75	1.34
金丝桃苷	98.26	96.88	98.59	97.91	0.93
迷迭香酸	99.57	102.86	97.24	99.89	2.83