高效液相色谱法测定口洁含漱液中13种化学成分的含量

doi:10.3969/j.issn.2097-2806.2023.10.016

摘要/Abstract

摘要：

建立了高效液相色谱法（HPLC）测定口洁含漱液中新绿原酸、绿原酸、咖啡酸、木犀草苷、3,5-O-二咖啡酰基奎宁酸、4,5-O-二咖啡酰基奎宁酸、木犀草素、芹菜素、马钱苷、当药苷、断氧化马钱子苷、（R,S）-告依春、直铁线莲宁B共13种成分的含量。以Waters Symmetry C₁₈（250 mm×4.6 mm，5 μm）为色谱柱，流动相：乙腈-0.3%磷酸，梯度洗脱，流速为1.0 mL/min，柱温为30 ℃，检测波长为348，240和224 nm。结果显示，口洁含漱液中13种化学成分的线性关系良好（r≥0.999 1），平均加样回收率为96.92%~100.11%，RSD为0.53%~1.58%。所建立的高效液相波长切换法结果准确，可用于口洁含漱液的质量控制。

关键词: 口洁含漱液, 高效液相色谱法, 口腔护理用品, 含量测定

Abstract:

The study aimed to develop an high performance liquid chromatography （HPLC） method for the determination of neochlorogenic acid, chlorogenic acid, caffeic acid, luteolin-7-O-glucoside, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, luteolin, apigenin, loganin, sweroside, secoxyloganin; （R, S）-goitrin and clemastanin B in Koujie gargle. Separation was carried out using Waters Symmetry C₁₈（250 mm×4.6 mm, 5 μm） with mobile phase of acetonitrile-0.3% phosphoric acid in gradient elution. The detection wavelength was set at 348 nm, 240 nm and 224 nm. The flow rate was 1.0 mL/min, and the column temperature was 30 ℃. The results show that the linear relationship of 13 chemical components in Koujie gargle is good （r≥0.999 1）, the average recovery ranges from 96.92% to 100.11%, and the RSD ranges from 0.53% to 1.58%. The established HPLC wavelength switching method is accurate which can be used for the quality control of Koujie gargle.

Key words: Koujie gargle, high performance liquid chromatography, oral care products, content determination

中图分类号:

TQ658

吴晓燕, 宋发祥. 高效液相色谱法测定口洁含漱液中13种化学成分的含量[J]. 日用化学工业（中英文）, 2023, 53(10): 1233-1240.

Wu Xiaoyan, Song Faxiang. Determination of 13 chemical components in Koujie gargle by HPLC[J]. China Surfactant Detergent & Cosmetics, 2023, 53(10): 1233-1240.

图/表 4

图1

表1

表2

加样回收试验结果（n=9）"

成分	样品中含量/mg	加入量/mg	测得量/mg	回收率/%	平均加样回收率/%	RSD/%
新绿原酸	0.361 0	0.285 6	0.639 6	97.55	99.02	1.08
	0.361 0	0.285 6	0.649 1	100.88
	0.361 0	0.285 6	0.646 4	99.93
	0.361 0	0.357 0	0.711 6	98.21
	0.361 0	0.357 0	0.710 8	97.98
	0.361 0	0.357 0	0.716 7	99.64
	0.361 0	0.428 4	0.783 1	98.53
	0.361 0	0.428 4	0.784 7	98.90
	0.361 0	0.428 4	0.787 6	99.58
绿原酸	5.929 0	4.715 2	10.593 3	98.92	98.21	1.1
	5.929 0	4.715 2	10.609 8	99.27
	5.929 0	4.715 2	10.612 0	99.32
	5.929 0	5.894 0	11.794 6	99.52
	5.929 0	5.894 0	11.701 5	97.94
	5.929 0	5.894 0	11.702 7	97.96
	5.929 0	7.072 8	12.785 9	96.95
	5.929 0	7.072 8	12.793 1	97.05
	5.929 0	7.072 8	12.784 3	96.92
咖啡酸	0.072 0	0.058 4	0.128 5	96.75	97.92	1.49
	0.072 0	0.058 4	0.130 1	99.49
	0.072 0	0.058 4	0.128 9	97.43
	0.072 0	0.073 0	0.142 3	96.30
	0.072 0	0.073 0	0.142 1	96.03
	0.072 0	0.073 0	0.144 9	99.86
	0.072 0	0.0876	0.159 1	99.43
	0.072 0	0.087 6	0.158 4	98.63
	0.072 0	0.087 6	0.157 3	97.37
木犀草苷	0.476 0	0.377 6	0.856 9	100.87	100.11	1.07
	0.476 0	0.377 6	0.843 7	97.38
	0.476 0	0.377 6	0.852 6	99.74
	0.476 0	0.472 0	0.949 4	100.30
	0.476 0	0.472 0	0.949 9	100.40
	0.476 0	0.472 0	0.950 8	100.59
	0.476 0	0.566 4	1.044 9	100.44
	0.476 0	0.566 4	1.046 5	100.72
	0.476 0	0.566 4	1.045 3	100.51
3,5-O-二咖啡酰基奎宁酸	1.216 0	0.967 2	2.168 1	98.44	98.22	1.27
	1.216 0	0.967 2	2.176 3	99.29
	1.216 0	0.967 2	2.160 9	97.69
	1.216 0	1.209 0	2.384 9	96.68
	1.216 0	1.209 0	2.381 3	96.39
	1.216 0	1.209 0	2.399 6	97.90
	1.216 0	1.450 8	2.671 4	100.32
	1.216 0	1.450 8	2.653 8	99.10
	1.216 0	1.450 8	2.640 5	98.19
4,5-O-二咖啡酰基奎宁酸	1.757 0	1.408 8	3.109 9	96.03	96.92	1.17
	1.757 0	1.408 8	3.110 7	96.09
	1.757 0	1.408 8	3.111 4	96.14
	1.757 0	1.761 0	3.469 1	97.22
	1.757 0	1.761 0	3.464 2	96.94
	1.757 0	1.761 0	3.450 6	96.17
	1.757 0	2.113 2	3.851 0	99.09
	1.757 0	2.113 2	3.836 5	98.41
	1.757 0	2.113 2	3.789 4	96.18
木犀草素	0.153 0	0.123 2	0.271 4	96.10	98.74	1.43
	0.153 0	0.123 2	0.274 9	98.94
	0.153 0	0.123 2	0.275 6	99.51
	0.153 0	0.154 0	0.306 3	99.55
	0.153 0	0.154 0	0.307 1	100.06
	0.153 0	0.154 0	0.306 4	99.61
	0.153 0	0.184 8	0.337 5	99.84
	0.153 0	0.184 8	0.334 6	98.27
	0.153 0	0.184 8	0.331 9	96.81
芹菜素	0.087 0	0.071 2	0.156 3	97.33	98.52	1.36
	0.087 0	0.071 2	0.157 9	99.58
	0.087 0	0.071 2	0.156 1	97.05
	0.087 0	0.089 0	0.174 2	97.98
	0.087 0	0.089 0	0.175 3	99.21
	0.087 0	0.089 0	0.173 0	96.63
	0.087 0	0.106 8	0.192 3	98.60
	0.087 0	0.106 8	0.193 7	99.91
	0.087 0	0.106 8	0.194 2	100.37
马钱苷	2.248 0	1.803 2	4.026 4	98.62	99.35	0.66
	2.248 0	1.803 2	4.031 5	98.91
	2.248 0	1.803 2	4.039 1	99.33
	2.248 0	2.254 0	4.516 7	100.65
	2.248 0	2.254 0	4.496 3	99.75
	2.248 0	2.254 0	4.495 6	99.72
	2.248 0	2.704 8	4.931 1	99.20
	2.248 0	2.704 8	4.912 0	98.49
	2.248 0	2.704 8	4.938 7	99.48
当药苷	0.687 0	0.553 6	1.229 4	97.98	98.97	1.22
	0.687 0	0.553 6	1.231 6	98.37
	0.687 0	0.553 6	1.224 7	97.13
	0.687 0	0.692 0	1.368 4	98.47
	0.687 0	0.692 0	1.379 3	100.04
	0.687 0	0.692 0	1.385 6	100.95
	0.687 0	0.830 4	1.517 1	99.96
	0.687 0	0.830 4	1.504 2	98.41
	0.687 0	0.830 4	1.512 7	99.43
断氧化马钱子苷	2.791 0	2.229 6	4.951 6	96.91	96.73	0.53
	2.791 0	2.229 6	4.954 3	97.03
	2.791 0	2.229 6	4.952 1	96.93
	2.791 0	2.787 0	5.470 9	96.16
	2.791 0	2.787 0	5.471 7	96.19
	2.791 0	2.787 0	5.469 1	96.09
	2.791 0	3.344 4	6.039 4	97.13
	2.791 0	3.344 4	6.019 2	96.53
	2.791 0	3.344 4	6.054 3	97.58
（R,S）-告依春	0.119 0	0.093 6	0.212 5	99.89	99.21	1.47
	0.119 0	0.093 6	0.210 4	97.65
	0.119 0	0.093 6	0.213 1	100.53
	0.119 0	0.117 0	0.235 3	99.40
	0.119 0	0.117 0	0.236 8	100.68
	0.119 0	0.117 0	0.235 7	99.74
	0.119 0	0.140 4	0.254 3	96.37
	0.119 0	0.140 4	0.257 1	98.36
	0.119 0	0.140 4	0.259 7	100.21
直铁线莲宁B	0.052 0	0.040 8	0.092 5	99.26	97.48	1.58
	0.052 0	0.040 8	0.091 4	96.57
	0.052 0	0.040 8	0.093 1	100.74
	0.052 0	0.051 0	0.101 3	96.67
	0.052 0	0.051 0	0.101 8	97.65
	0.052 0	0.051 0	0.101 1	96.27
	0.052 0	0.061 2	0.111 2	96.73
	0.052 0	0.061 2	0.110 9	96.24
	0.052 0	0.061 2	0.111 5	97.22

表2

表3

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组分	回归方程	r	线性范围/（μg/mL）
新绿原酸	Y=9.841 6×10⁵X-813.9	0.999 6	2.39~119.50
绿原酸	Y=1.351 2×10⁶X+1 543.1	0.999 2	40.65~2 032.50
咖啡酸	Y=5.982 4×10⁵X-1 813.6	0.999 3	0.49~24.50
木犀草苷	Y=1.523 9×10⁶X+613.8	0.999 1	3.06~153.00
3,5-O-二咖啡酰基奎宁酸	Y=6.931 8×10⁵X-961.4	0.999 9	7.95~397.50
4,5-O-二咖啡酰基奎宁酸	Y=1.109 7×10⁶X-846.2	0.999 4	11.88~594.00
木犀草素	Y=1.485 1×10⁶X-613.7	0.999 2	1.04~52.00
芹菜素	Y=8.330 6×10⁵X+516.5	0.999 3	0.56~28.00
马钱苷	Y=1.902 4×10⁶X-1 064.2	0.999 5	16.02~801.00
当药苷	Y=1.722 5×10⁶X-436.8	0.999 7	4.35~217.50
断氧化马钱子苷	Y=2.185 7×10⁶X+491.6	0.999 6	19.59~979.50
（R,S）-告依春	Y=1.215 9×10⁶X+1 162.3	0.999 8	0.77~38.50
直铁线莲宁B	Y=3.157 4×10⁵X-899.1	0.999 4	0.36~18.00

批号	含量/（mg/mL）
批号	新绿原酸	绿原酸	咖啡酸	木犀草苷	3,5-O-二咖啡酰基奎宁酸	4,5-O-二咖啡酰基奎宁酸	木犀草素	芹菜素	马钱苷	当药苷	断氧化马钱子苷	（R,S）-告依春	直铁线莲宁B
H2073	0.361	5.929	0.072	0.476	1.216	1.757	0.153	0.087	2.248	0.687	2.791	0.119	0.052
H2111	0.346	5.750	0.069	0.463	1.162	1.691	0.155	0.086	2.691	0.671	2.696	0.115	0.050
H2116	0.379	6.183	0.078	0.504	1.318	1.907	0.164	0.091	2.418	0.734	2.981	0.127	0.054