高效液相色谱法测定追风活血膏中8种成分

doi:10.3969/j.issn.1001-1803.2022.12.012

摘要/Abstract

摘要：

建立了高效液相色谱（HPLC）测定追风活血膏中士的宁、乌头碱、马钱子碱、当归、乳香、没药、蜕皮甾酮和青藤碱8种成分含量的方法。色谱柱为Agilent ZORBAXSB-C₁₈柱（4.6 mm×250 mm, 5 μm）；流动相：甲醇（A相），0.1% H₃PO₄水溶液（B相）；流速1.0 mL/min；柱温室温；检测波长256 nm。结果显示，8种成分能够达到良好分离，在各自线性范围内线性关系良好，相关系数r均大于0.999；平均加样回收率在98.74%~99.67%之间，相对标准偏差（RSD）在0.85%~1.76%范围内。5批次样品中士的宁、乌头碱、马钱子碱、当归、乳香、没药、蜕皮甾酮和青藤碱的含量分别为0.031~0.349，0.021~0.166，0.024~0.298，0.021~0.219，0.017~0.186，0.015~0.158，0.015~0.149，0.016~0.179 mg/g。本文所建立的HPLC法操作简单、稳定可控，可用于追风活血膏的质量控制。

关键词: 追风活血膏, 高效液相色谱法, 质量控制

Abstract:

A high performance liquid chromatographic （HPLC） method was developed for determination of the eight components in Zhuifeng Huoxue Ointment, namely, brucine, aconitine, strychnine, angelica, frankincense, myrrh, ecdysterone and sinomenine. The chromatographic column was an Agilent ZORBAXSB-C₁₈ column （4.6 mm×250 mm, 5 μm）; the mobile phase was methanol （phase A） and 0.1% aqueous solution of H₃PO₄ （phase B）; the flow rate was 1.0 mL/min; the column temperature was room temperature; the detection wavelength was 256 nm. The average recoveries ranged from 98.74%-99.67%, and the relative standard deviations （RSDs） were in the range of 0.85%-1.76%. The content of strychnine, aconitine, brucine, angelica, frankincense, myrrh, ecdysterone and sinomenine in the five batches of samples were 0.031-0.349, 0.021-0.166, 0.024-0.298, 0.021-0.219, 0.017-0.186, 0.015-0.158, 0.015-0.149, 0.016-0.179 mg/g, respectively. The HPLC method developed is simple, stable and controllable, and can be used for the quality control of Zhuifeng Huoxue Ointment.

Key words: Zhuifeng Huoxue Ointment, high performance liquid chromatography, quality control

中图分类号:

TQ658

万刘静,张利. 高效液相色谱法测定追风活血膏中8种成分[J]. 日用化学工业（中英文）, 2022, 52(12): 1342-1347.

Wan Liujing,Zhang Li. Determination of 8 components in Zhuifeng Huoxue Ointment by high performance liquid chromatography[J]. China Surfactant Detergent & Cosmetics, 2022, 52(12): 1342-1347.

图/表 4

图1

表1

表2

表3

参考文献 17

[1]	Xu Minmin, Liu Shaofan, Wang Dingju. Clinical observation on acute gouty arthritis (damp-heat arthritis syndrome) treated by acupoint application of zhongyi plaster[J]. China Emergency in Traditional Chinese Medicine, 2018, 27 (1) : 112-114.
[2]	Wang Chuanbang, Liu Tingting, Mou Liqiu, et al. Research progress and application prospects of a new microemulsion drug delivery system[J]. China Pharmaceuticals, 2016, 25 (1) : 5-10.
[3]	Xu Bo, Zhang Laigen, Zhang Nongshan, et al. Clinical study on 372 cases of knee osteoarthritis treated with “mems microneedle penetration promoting’easy layer’ application technique”[J]. Shizhen Medicine and Materia Medica Research, 2016, 27 (8) : 1912-1914.
[4]	Liu Di, Zhang Bingyang, Yao Tie, et al. Research progress on the chemical constituents and pharmacological effects of Boswellia serrata[J]. Chinese Herbal Medicine, 2020, 51 (22) : 5900-5914.
[5]	Zhang Haodong, Dou Hongli, Ye Mengyi. A study on the herbal literature of myrrh’s properties and efficacy application[J]. Shanxi Journal of Traditional Chinese Medicine, 2020, 36 (3) : 49-57.
[6]	Li Shuang, Li Rui, Zeng Yong, et al. Research progress on the chemical constituents and pharmacological effects of chuanwu[J]. China Journal of Chinese Materia Medica, 2019, 44 (12) : 2433-2443. doi: 10.19540/j.cnki.cjcmm.20190221.004 pmid: 31359708
[7]	Yao Nan, Wang Zhiwang, Fu Xiaoyan, et al. Research progress on the effect of angelica volatile oil and its phthalides on smooth muscle[J]. The Chinese Journal of Modern Applied Pharmacy, 2019, 36 (21) : 2738-2742.
[8]	Fan Jian, Shen Ying, Zhao Juanping, et al. Application status and research progress of topical Chinese medicinal ointments for rheumatism treatment[J]. Chinese Journal of Experimental Formulas, 2019, 25 (2) : 206-213.
[9]	Wang Dandan, Li Wuyin. Existing problems of black plaster preparation process and the status quo of improvement research[J]. China Medical Herald, 2019, 16 (4) : 30-33.
[10]	Zhang Bohua, Zhang Yisheng, Li Jie, et al. Study on the preparation process of black plaster for children with cough and asthma[J]. China Pharmacist, 2015, 18 (2) : 332-334.
[11]	Yin Shanshan, Wang Yanqiu, Zhou Guoliang, et al. Effects of different penetration enhancers on the in vitro transdermal absorption of naringin in sanwei dieda fengshi plaster[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2016, 22 (7) : 15-18.
[12]	Jiang Minyu, Yan Dan, Chen Jiao, et al. Preparation of sanqi dieda ointment and its in vitro transdermal properties[J]. Chinese and Herbal Medicine, 2017, 48 (22) : 4639-4647.
[13]	Ma Zhenyong, Li Yukui, He Jinfeng, et al. Experimental study on long-term skin toxicity of compound aconitum microemulsion in rats[J]. Journal of Chinese Medicine, 2017, 32 (12) : 2423-2425.
[14]	Yuan Weibin, Wu Han, Huang Xiaobing, et al. Study on the safety evaluation of the new process wentong ointment[J]. Chinese Materia Medica, 2017, 40 (8) : 1956-1965.
[15]	Zeng Zhaoyi, Liu Jia, Xiao Fang, et al. The effect of three ways of adding volatile oil on the matrix and in vitro release of analgesic gel patch[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2017, 23 (15) : 20-25.
[16]	Yuan Jie, Liu Daofang. Study on the quality standard of jieguling plaster[J]. Chinese Patent Medicine, 2016, 38 (3) : 575-578.
[17]	Duan Jijiang, Li Shasha, Weng Lidong, et al. Determination of berberine hydrochloride in tianqi dieda fengshi ointment by HPLC[J]. Chinese Journal of Experimental Formulas, 2013, 19 (13) : 85-87.

成分	回归方程	线性范围/（μg/mL）	r
士的宁	Y=15.014+274.516X	0.059~0.592	0.999 9
乌头碱	Y=732.026-15.781X	0.131~1.310	0.999 9
马钱子碱	Y=167.127+478.137X	0.118~1.180	0.999 8
当归	Y=237.514+115.241X	0.071~0.714	0.999 9
乳香	Y=72.241+173.320X	0.079~0.792	0.999 8
没药	Y=632.843+392.173X	0.091~0.914	0.999 9
蜕皮甾酮	Y=184.243+112.834X	0.174~1.740	0.999 8
青藤碱	Y=461.367+228.258X	0.068~0.684	0.999 9

成分	称取量/g	样品中含量/mg	加入量/mg	测得量/mg	回收率/%	平均回收率/%	RSD/%
士的宁	2.048	0.332	0.334	0.666	99.18	99.31	0.17
	2.194	0.337	0.334	0.671	99.46
	2.004	0.338	0.334	0.672	99.52
	1.991	0.333	0.334	0.667	99.09
	1.994	0.335	0.334	0.669	99.35
	1.993	0.330	0.334	0.663	99.26
乌头碱	2.018	0.185	0.185	0.370	98.54	98.74	0.19
	2.015	0.181	0.185	0.366	98.68
	2.020	0.190	0.185	0.375	98.97
	1.995	0.190	0.185	0.374	98.69
	1.991	0.182	0.185	0.367	98.62
	1.990	0.182	0.185	0.367	99.04
马钱子碱	2.020	0.214	0.214	0.428	99.18	99.25	0.12
	2.012	0.214	0.214	0.428	99.34
	2.095	0.216	0.214	0.430	99.76
	2.038	0.215	0.214	0.429	99.09
	1.999	0.213	0.214	0.427	98.93
	1.998	0.212	0.214	0.426	99.79
当归	2.017	0.102	0.103	0.205	99.84	99.67	0.17
	2.028	0.102	0.103	0.205	99.15
	2.075	0.103	0.103	0.206	99.32
	2.086	0.106	0.103	0.209	99.84
	1.997	0.105	0.103	0.208	99.69
	1.991	0.100	0.103	0.203	100.18
乳香	2.083	0.082	0.084	0.166	99.14	99.36	0.15
	2.049	0.086	0.084	0.169	99.51
	2.094	0.082	0.084	0.166	99.98
	1.996	0.084	0.084	0.167	99.09
	1.999	0.085	0.084	0.168	99.12
	1.991	0.083	0.084	0.167	99.32
没药	2.020	0.090	0.092	0.181	98.74	98.79	0.14
	2.097	0.091	0.092	0.183	98.57
	2.047	0.093	0.092	0.185	99.13
	2.096	0.095	0.092	0.186	98.21
	1.993	0.092	0.092	0.184	98.85
	1.996	0.089	0.092	0.181	99.24
蜕皮甾酮	1.991	0.090	0.088	0.178	98.47	99.18	0.21
	1.998	0.087	0.088	0.175	99.13
	1.997	0.086	0.088	0.175	99.42
	1.998	0.087	0.088	0.175	99.97
	2.017	0.090	0.088	0.178	98.85
	2.085	0.091	0.088	0.179	99.24
青藤碱	2.066	0.147	0.151	0.298	99.17	99.54	0.16
	2.016	0.152	0.151	0.303	99.51
	1.999	0.152	0.151	0.303	99.56
	1.995	0.150	0.151	0.301	99.74
	1.995	0.150	0.151	0.301	99.38
	1.999	0.155	0.151	0.306	99.88

成分	含量/（mg/g）
	批号100801		批号100903		批号101101		批号101201		批号101205
	醇提	油炸	醇提	油炸	醇提	油炸	醇提	油炸	醇提	油炸
士的宁	0.343	0.032	0.348	0.031	0.345	0.032	0.349	0.031	0.346	0.031
乌头碱	0.166	0.021	0.165	0.021	0.161	0.021	0.159	0.021	0.164	0.021
马钱子碱	0.297	0.025	0.291	0.025	0.294	0.024	0.298	0.024	0.292	0.025
当归	0.215	0.022	0.211	0.021	0.210	0.021	0.219	0.022	0.216	0.022
乳香	0.186	0.018	0.184	0.017	0.182	0.017	0.187	0.017	0.185	0.018
没药	0.151	0.016	0.157	0.016	0.154	0.015	0.152	0.015	0.158	0.015
蜕皮甾酮	0.148	0.015	0.141	0.016	0.142	0.016	0.149	0.015	0.144	0.016
青藤碱	0.172	0.017	0.170	0.016	0.179	0.016	0.175	0.017	0.176	0.016