灯盏花化妆品原料的抗氧化活性及质量控制研究

doi:10.3969/j.issn.1001-1803.2021.12.006

摘要/Abstract

摘要：

对灯盏花化妆品原料（EBCI）的抗氧化活性进行研究,同时建立其中两种指标成分的含量测定方法。采用DPPH、ABTS、·OH清除法考察EBCI的自由基清除活性,同时用H₂O₂诱导HHF-1细胞氧化损伤模型考察EBCI对细胞存活率和细胞中谷胱甘肽（GSH）含量的影响;建立了HPLC法同时测定EBCI中野黄芩苷和4,5-二-O-咖啡酰奎宁酸的含量测定方法。结果显示EBCI具有DPPH、ABTS和·OH自由基清除能力,IC₅₀分别为46.93、91.36和89.37 μg/mL,在4、20 μg/mL质量浓度下能显著提高氧化损伤HFF-1细胞的存活率且能显著提高细胞中GSH含量。野黄芩苷和4,5-二-O-咖啡酰奎宁酸分别在0.009 3~0.372 2、0.008 5~0.340 9 mg/mL范围内呈现良好线性关系,加样回收率分别为103.47%、103.38%（RSD均小于2%）。结果表明EBCI具有抗氧化作用;所建立的含量测定方法简便、稳定、准确可靠,可用于EBCI质量控制。

关键词: 灯盏花, 化妆品原料, 抗氧化, 野黄芩苷, 4,5-二-O-咖啡酰奎宁酸, 含量测定

Abstract:

Erigeron breviscapus （vant.） Hand. Mazz. is one of the most important traditional Chinese medicinal plants for cardiopathy. Erigeron breviscapus mainly contains flavonoids and caffeic acids, which have obvious antioxidant activities. To study the possibility of developing Erigeron breviscapus as a cosmetics material, the antioxidant activity of Erigeron breviscapus cosmetic ingredients （EBCI） was studied and a method for simultaneous determination of two index compounds therein was established. The scavenging activities of EBCI against DPPH, ABTS and ·OH radicals were evaluated by in vitro method. The effects of EBCI on cell viability and glutathione （GSH） content were investigated by using H₂O₂-induced damage in HHF-1 cells. HPLC method for simultaneous determination of scutellarin and 4,5-dicaffeoylquinic acid in EBCI was established. The results show that EBCI have scavenging activities against DPPH, ABTS and ·OH radicals, with IC₅₀ values being 46.93, 91.36 and 89.37 μg/mL, respectively. The cell viability of H₂O₂-induced injury in HFF-1 cells and the GSH content in the cells could be significantly increased at the concentrations of 4 and 20 μg/mL. 4,5-Dicaffeoylquinic acid and scutellarin showed good linearity in the linear ranges of 0.008 5-0.340 9 mg/mL and 0.009 3-0.372 2 mg/mL, respectively. Average recoveries of scutellarin and 4,5-dicaffeoylquinic acid were 103.47% and 103.38% （RSD<2%）, respectively. This method is accurate, simple and well reproducible, and can be used for the quality control of EBCI. It shows the antioxidant activity of EBCI in this study.

Key words: Erigeron breviscapus, cosmetic ingredient, antioxidant, scutellarin, 4,5-dicaffeoylquinic acid, content determination

中图分类号:

TQ658

李倩,徐明,肖茹月,罗春,李战国,梅双喜. 灯盏花化妆品原料的抗氧化活性及质量控制研究[J]. 日用化学工业, 2021, 51(12): 1202-1207.

Li Qian,Xu Ming,Xiao Ruyue,Luo Chun,Li Zhanguo,Mei Shuangxi. Antioxidant activity analysis and quality control of cosmetic ingredients from Erigeron breviscapus[J]. China Surfactant Detergent & Cosmetics, 2021, 51(12): 1202-1207.

图/表 6

表 1

表 2

表 3

图 1

表 4

表 5

参考文献 10

[1]	Zhang Qihui, Zhang Li, Niu Huanmin, et al. Erigeron breviscapus preparation in cerebrovascular disease[J]. Jilin Journal of Traditional Chinese Medicine, 2016, 36(9) : 941-943.
[2]	Lu Yizhong, Li Hehua, Lu Yifan, et al. Fleabane combined with bone marrow mesenchymal stem cell transplantation for cerebral infarction[J]. Chinese Journal of Tissue Engineering Research, 2015, 44(50) : 8114-8119.
[3]	Zhou Ling, Xie Liyan, Xu Jie, et al. Simultaneous determination of six major compounds in dengzhanxixin injection by HPLC[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2011, 17(21) : 78-81.
[4]	Chen Chuping, Wang Jianzhuang, Zhou Ye. Determination of scutellarin in erigerontis herba granules[J]. Journal of Guangdong Pharm Aceutical College, 2010, 26(3) : 279-282.
[5]	Zhu Fayan, Yang Fan, Wang Yajun, et al. Research progress of flavonoids in erigerontig herba[J]. Journal of Anhui Agriculture Sciences, 2012, 40(10) : 5853-5857.
[6]	Zhang Jing, Wang Yujie, Pang Yuan, et al. The CE-CL finger prints reseach of antioxidant activity effective components of erigeron injection[J]. Chinese Journal of Ethnomedicine and Ethnopharmacy, 2011: 69-71.
[7]	Ma Li, Liao Yanhua, Han Feng, et al. Determination of scutellarin and 4,5-dicaffeoylquinic acid in Xinnao Luotong Granulesby HPLC[J]. Drugs & Clinic, 2012, 27(6) : 582-584.
[8]	Liu Hong, Yang Xiangliang, Zhou Lizhu, et al. Study on effects scutellarin on scavenging reactive oxygen[J]. Journal of Chinese Medicinal Materials, 2002, 25(7) : 491-493. pmid: 12599762
[9]	Hong Hong, Liu Guoqing. Protection against hydrogen peroxide-induced Cytotoxitity in PC 12 cells by scutellarin[J]. Life Sciences, 2004, 74(24) : 2989-2973.
[10]	Liu Hong, Yang Xiangliang, Wang Ying, et al. Protective effects of scutellarin on superoxide-induced oxidative stress in rats cortical synapto-somes[J]. Acta Pharmacologica Sincia, 2003, 24(11) : 1113-1117.

时间/min	φ（流动相A）/%	φ（流动相B）/%
0~10	95→88	5→12
10~15	95→85	12→15
15~30	85→84	15→16
30~40	84	16
40~55	84→83	16→17
55~60	83→10	17→90

样品	DPPH		ABTS		·OH
样品	ρ/（μg·mL^-1）	清除率/%	ρ/（μg·mL^-1）	清除率/%	ρ/（μg·mL^-1）	清除率/%
EBCI	125	87.91±0.28	250	91.90±1.22	250	85.23±2.69
	62.5	62.82±7.10	125	65.36±0.94	125	60.93±5.59
	31.25	31.79±0.16	62.5	31.41±0.28	62.5	38.36±4.91
	15.625	17.10±1.19	31.25	18.41±0.19	31.25	24.35±1.66
Vc	20.0	93.43±1.46	20.0	83.81±1.98	125	81.81±3.78
	10.0	66.96±12.83	10.0	35.12±2.85	62.5	53.95±3.21
	5.0	35.57±4.08	5.0	20.54±2.09	31.25	37.41±0.57
	2.5	19.92±3.78	2.5	14.17±3.58	15.625	28.86±5.97

组别	ρ/（μg·mL^-1）	细胞存活率/%	GSH含量/（μmol·gprot^-1）
空白对照组	—	100.00±1.08	20.73±4.21
模型组	—	35.79±2.41^###	13.53±1.60^#
EBCI	100	41.24±2.85	19.54±1.51^*
	20	42.43±3.13^*	20.47±3.15^*
	4	41.51±1.21^*	23.63±3.42^*
V_E	100	92.63±11.56^***	20.52±3.61^*
	20	50.67±7.07^*	19.85±2.24^*
	4	34.88±4.11	—

对照品	称样量/mg	样品中含量/mg	加入量/mg	测得量/mg	回收率/%	平均值/%	RSD值/%
野黄芩苷	10.4	0.359 5	0.348 9	0.719 2	103.08	103.47	0.58
	10.2	0.352 6	0.348 9	0.713 8	103.52
	10.7	0.369 9	0.348 9	0.733 3	104.15
	10.6	0.366 5	0.348 9	0.724 4	102.58
	10.3	0.356 1	0.348 9	0.719 2	104.08
	10.5	0.363 0	0.348 9	0.723 9	103.43
4,5-二-O-咖啡酰奎宁酸	10.0	0.145 4	0.127 8	0.280 0	105.30	103.38	1.58
	10.3	0.136 9	0.127 8	0.267 3	102.04
	10.7	0.141 7	0.127 8	0.271 8	101.77
	10.4	0.126 0	0.127 8	0.256 4	102.00
	10.1	0.122 3	0.127 8	0.255 5	104.16
	10.7	0.129 6	0.127 8	0.263 9	105.04

样品批号	w（野黄芩苷）/ %	w（4,5-二-O-咖啡酰奎宁酸）/ %
EBCI-1	3.52	1.26
EBCI-2	3.65	1.40
EBCI-3	3.36	1.40
EBCI-4	3.23	1.00
EBCI-5	3.72	1.11