归芩牙膏中阿魏酸、黄芩苷含量测定及其药效学研究

doi:10.3969/j.issn.2097-2806.2023.11.015

摘要/Abstract

摘要：

建立高效液相色谱（HPLC）测定归芩牙膏中阿魏酸、黄芩苷含量的方法，研究其抗炎、镇痛、止血等药理作用。通过HPLC对归芩牙膏中君药当归、黄芩的主要成分阿魏酸、黄芩苷进行含量测定，色谱柱为Agilent ZOBAX SB C₁₈（2.1 mm×150 mm，5 μm），流动相为乙腈-0.2%乙酸水溶液（梯度洗脱），流速为1.0 mL/min，检测波长为320 nm，柱温为35 ℃。通过建立二甲苯致小鼠耳肿胀模型、小鼠热板法和小鼠肝脏创面出血模型考察归芩牙膏抗炎、镇痛及止血作用。并通过ELISA法检测小鼠耳廓组织中肿瘤坏死因子-α （TNF-α）、白介素-1β （IL-1β）、白介素-6 （IL-6）等炎症因子的表达情况。阿魏酸、黄芩苷进样量分别在3.04~304 μg/mL，2.86~286 μg/mL范围内进样质量浓度与峰面积均呈现良好的线性关系（R²>0.999 7）；精密度、稳定性、重复性试验RSD均小于3.0%；平均加样回收率分别为98.95%，99.87%，RSD分别为1.51%，0.22%。样品中每1 g含阿魏酸应不低于0.54 mg，含黄芩苷应不低于16.8 mg。HPLC法专属性强，重复性和稳定性良好，可用于归芩牙膏的质量控制。药效学实验表明归芩牙膏具有良好的抗炎、镇痛和止血作用。本研究为后期归芩牙膏的开发及临床应用提供了实验依据。

关键词: 归芩牙膏, 阿魏酸, 黄芩苷, 含量测定, 高效液相色谱

Abstract:

A method for determining the content of ferulic acid and baicalein in the toothpaste by high performance liquid chromatography （HPLC） was established, and its pharmacological effects such as anti-inflammatory, analgesic and hemostatic effects were studied. Angelica and Baical skullcap root are the main medicines of Gui Qin toothpaste. The content of ferulic acid and baicalein, the main components of Gui Qin toothpaste, was determined by HPLC. The column was Agilent ZOBAX SB C₁₈ （2.1 mm×150 mm, 5 μm）, the mobile phase was acetonitrile-0.2% acetic acid aqueous solution （gradient elutation）, the flow rate was 1.0 mL/min, the detection wavelength was 320 nm, and the column temperature was 35 ℃. The anti-inflammatory, analgesic and hemostatic effects of Gui Qin toothpaste were investigated by establishing a model of xylene-induced ear swelling, mouse hot plate method and mouse liver wound bleeding model. The expression of inflammatory factors such as tumor necrosis factor-α （TNF-α）, interleukin-1β （IL-1β） and interleukin-6 （IL-6） in the mouse ear tissues was detected by ELISA. The injection rates of ferulic acid and baicalin show a good linear relationship between the injection concentration and the peak area in the range of 3.04-304 μg/mL and 2.86-286 μg/mL, respectively （R²>0.999 7）. The RSDs of the precision, stability and repeatability tests are less than 3.0%. The average dosing recovery rates are 98.95% and 99.87%, and the RSDs are 1.51% and 0.22%, respectively. The sample should contain no less than 0.54 mg of ferulic acid per 1 g and no less than 16.8 mg of baicalin. The HPLC method has strong specificity, good repeatability and stability, and can be used for quality control of Gui Qin toothpaste. Pharmacodynamic experiments have shown that Gui Qin toothpaste has good anti-inflammatory, analgesic and hemostatic effects. This study provides an experimental basis for the development and clinical application of Gui Qin toothpaste in the later stage.

Key words: Gui Qin toothpaste, ferulic acid, baicalein, content determination, high performance liquid chromatography

中图分类号:

TQ658

郑丽霞, 高慧琴, 李飒, 郭从嘉, 郝倩莹, 吴国泰. 归芩牙膏中阿魏酸、黄芩苷含量测定及其药效学研究[J]. 日用化学工业（中英文）, 2023, 53(11): 1345-1350.

Zheng Lixia, Gao Huiqin, Li Sa, Guo Congjia, Hao Qianying, Wu Guotai. Determination of ferulic acid and baicalin content in Gui Qin toothpaste and its pharmacodynamic study[J]. China Surfactant Detergent & Cosmetics, 2023, 53(11): 1345-1350.

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参考文献 11

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时间/min	φ（流动相A）/%	φ（流动相B）/%
0~ 2	13	87
2~18	13→30	87→70
18~25	30	70
25~28	30→13	70→87
28~30	13	87

成分	样品含量/mg	加入量/mg	测得量/mg	加样回收率/%	平均加样回收率/%	RSD/%
阿魏酸	0.053 6	0.04	0.093 0	98.50	98.95	1.51
	0.054 0	0.04	0.092 6	96.50
	0.054 2	0.04	0.093 8	99.00
	0.052 4	0.05	0.102 8	100.80
	0.053 6	0.05	0.102 0	96.80
	0.054 2	0.05	0.104 0	99.60
	0.053 6	0.06	0.114 0	100.67
	0.052 4	0.06	0.112 0	99.33
	0.053 0	0.06	0.112 6	99.33
黄芩苷	1.685 6	1.34	3.026 0	100.03	99.87	0.22
	1.680 2	1.34	3.016 6	99.73
	1.690 6	1.34	3.022 0	99.36
	1.680 2	1.68	3.359 0	99.93
	1.690 6	1.68	3.369 0	99.91
	1.679 8	1.68	3.360 2	100.02
	1.680 2	2.01	3.686 0	99.79
	1.681 4	2.01	3.690 0	99.93
	1.685 6	2.01	3.697 4	100.09

成分	阿魏酸峰面积/（mAU·min）	ρ（阿魏酸）/（μg/mL）	阿魏酸含量/%	黄芩苷峰面积/（mAU·min）	ρ（黄芩苷）/（μg/mL）	黄芩苷含量/%
1	847.6	7.35	0.054	9 480.9	227.19	1.68
2	847.0	7.35		9 479.6	227.16
3	846.4	7.34		9 479.5	227.16

组别	剂量	耳廓肿胀度	肿胀抑制率/%
空白组	0.2 mL NS	—	—
模型组	0.2 mL NS	8.25±1.80	—
阳性对照组	0.2 g	4.63±1.56^**Δ	43.94
牙膏基质组	0.2 g	6.09±0.92^*	26.21
归芩牙膏组	0.2 g	4.11±1.54^**ΔΔ	50.15

组别	剂量	ρ（TNF-α）/（ng/L）
空白组	0.2 mL NS	932.5 ±20.41
模型组	0.2 mL NS	1 178.85±37.83^**
阳性对照组	0.2 g	1 098.65±67.75
牙膏基质组	0.2 g	1 158.02±94.08
归芩牙膏组	0.2 g	1 068.96±48.32^ΔΔ