高效液相色谱法测定化妆品中14种α-羟基酸和羟基酸酯

doi:10.3969/j.issn.2097-2806.2024.03.015

摘要/Abstract

摘要：

采用高效液相色谱法（HPLC）建立同时测定化妆品中14种α-羟基酸和羟基酸酯含量的方法，并结合高效液相色谱-串联质谱联用法（HPLC-MS/MS）确证结果。采用高效液相色谱法，以0.05 mol/L的磷酸氢二铵水溶液和甲醇为流动相进行梯度洗脱，采用Waters T3-C18 （4.6×250 mm，5 µm）色谱柱分离，采用二极管阵列检测器检测，以214 nm为检测波长，外标法定量。采用高效液相色谱-串联质谱联用法，以0.1%甲酸水溶液和含0.1%甲酸乙腈为流动相进行梯度洗脱，采用Waters T3-C18 （2.1×100 mm，1.8 µm）色谱柱分离，电喷雾电离，正、负离子多反应监测模式定性。14种化合物在各自质量浓度范围内呈良好的线性关系，相关系数R²均大于0.999。4种基质样品，14种化合物平均加标回收率均在85%~115%范围内，相对标准偏差均小于5%，检出限为1~150 μg/g，定量限为3~450 μg/g。对7批含α-羟基酸和酯的样品进行液质确证，结果均为阳性。该方法操作简便、准确度好、灵敏度高，适用于化妆品中14种α-羟基酸及其酯的测定。

关键词: 化妆品, α-羟基酸, 乳酸甲酯, 乳酸乙酯, 乳酸丁酯, 高效液相色谱法

Abstract:

The study aimed to establish a method for simultaneous determination of 14 kinds of α-hydroxy acids and their esters in cosmetics by high performance liquid chromatography （HPLC），and the results were confirmed by high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）. HPLC was used for gradient elution with 0.05 mol/L diammonium hydrogen phosphate-methanol as the mobile phase. The analytes were separated on a Waters T3-C18 （4.6×250 mm, 5 µm） column and detected at the wavelengths of 214 nm. Quantification analysis was performed by the external standard method. HPLC-MS/MS was used for gradient elution with 0.1% formic acid-acetonitrile containing 0.1% formic acid as the mobile phase, and the analytes were separated on a Waters T3-C18 （2.1×100 mm, 1.8 µm） column. Qualitative analysis was performed by electronic spray ionization under a positive ion and negative ion mode with multiple reaction monitoring （MRM）. The linearity of the 14 components in the concentration range is good, with the correlation coefficient r≥0.999. The average spiked recoveries of 14 compounds in 4 cosmetic matrices are all in the range of 85%-115%, and the relative standard deviations are all less than 5%. The limits of detections range from 1 μg/g to 150 μg/g. The limits of quantitation range from 3 μg/g to 450 μg/g. Seven batches of samples containing α-hydroxy acid and its ester were confirmed by HPLC-MS/MS and the results are positive. The results indicate that the method is simple, sensitive and accurate, and is suitable for the determination of 14 kinds of α-hydroxy acids and their esters in cosmetics.

Key words: cosmetics, α-hydroxy acids, methyl lactate, ethyl lactate, butyl lactate, high performance liquid chromatography （HPLC）

中图分类号:

TQ658

张丽媛, 颜琳琦, 程巧鸳, 戚绿叶, 王容, 黄柳倩. 高效液相色谱法测定化妆品中14种α-羟基酸和羟基酸酯[J]. 日用化学工业（中英文）, 2024, 54(3): 353-359.

Liyuan Zhang, Linqi Yan, Qiaoyuan Cheng, Lvye Qi, Rong Wang, Liuqian Huang. Determination of 14 kinds of α-hydroxy acids and hydroxy esters in cosmetics[J]. China Surfactant Detergent & Cosmetics, 2024, 54(3): 353-359.

图/表 6

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参考文献 14

[1]	Liang Jiaqi. Summary of patented technologies for the application of α-hydroxy acids in cosmetics[J]. Shandong Chemical Industry, 2020, 49 (14) : 70-71, 76.
[2]	Li Yangjie, Wu Zhen, Xiao Shuxiong, et al. Simultaneous determination of 10 α-hydroxy acids in cosmetics by high performance liquid chromatography[J]. Chin J Health Lab Tec, 2017, 27 (19) : 2757-2760.
[3]	Zuo Xue, Han Xiaoqian, Liu Chenghao, et al. Determination of ten kinds of α-hydroxy acids in cosmetics with quantitative analysis of multi-components by single marker[J]. Journal of Hygiene Research, 2020, 49 (2) : 285-289, 319. doi: 10.19813/j.cnki.weishengyanjiu.2020.02.020 pmid: 32290947
[4]	He Qiaosang, Lu Yan. Determinnation of 6 kinds of organic acid in cosmetics by high performance liquid chromatography[J]. Chemical Industry Times, 2016, 19 (5) : 991-993.
[5]	Meng Huimin, Li Li. The effect and clinical application of fruit acid[J]. J Dermatology and Venereology, 2014, 36 (3) : 155-157.
[6]	Standardization Administration. Safety and technical standards for cosmetics[S]. Beijing: Standards Press of China, 2016.
[7]	Zhang Hong, Wang Bin, Zhou Shiwei. Determination of α-hydroxy acids (AHAs) in cosmetics by gas chromatography[J]. Sh J Prev Med, 2002, 14 (6) : 261-262.
[8]	Zhong Zhixiong, Du Daan, Liang Xuxia. Determination of α-hydroxy acids in cosmetics by iron chromatography[J]. Chin J Health Lab Tec, 2001, 11 (1) : 21-26.
[9]	Zhong Zhixiong, Liang Chunshui, Du Daan. Determination of organic acids in cosmetics by ion-exchange chromatography suppressed conductivity detection[J]. J Environ Health, 2006, 23 (4) : 357-360.
[10]	Zhu Huijuan, Liu Siyang, Yan Yanwei, et al. Comparison of detection methods of α-hydroxy acids in cosmetics and disinfectants[J]. J Environ Health, 2014, 31 (3) : 267-268.
[11]	Moona K, Limb C, Kimb S, et al. Facile determination of the absolute configurations of α-hydroxy acids by chiral derivatization coupled with liquid chromatography-mass spectrometry analysis[J]. J Chromatogr A, 2013, 1272: 141-144. doi: 10.1016/j.chroma.2012.11.006
[12]	Fransson B, Ragnarsson U. Separation of enantiomers of α-hydroxy acids by reversed-phase liquid chromatography after derivatization with 1-(9-fluorenyl) ethyl chloroformat[J]. J Chromatogr A, 1998, 827: 31-36. doi: 10.1016/S0021-9673(98)00770-5
[13]	Huang An, Zeng Limei, Feng Xuehua, et al. Application research of the natural organic acid to the vesicant and thickener in the cosmetic industry[J]. Journal of Changsha University of Electric Power (Natural Science), 2003, 18 (3) : 84-85.
[14]	National Medical Products Administration. Announcement of the National Medical Products Administration on the Issuance of the Catalogue of Used Cosmetic Ingredients (2021 Edition) (No. 62, 2021) [EB/OL].(2021)[2023-03-10]. https://www.nmpa.gov.cn/directory/web/nmpa/xxgk/ggtg/qtggtg/20210430162707173.html.

序号	组分名称	母离子（m/z）	子离子（m/z）	碰撞能/V
1	葡糖醛酸	193.2	113.1	-14
1	葡糖醛酸	193.2	131.1	-11
2	酒石酸	149.1	87.1	-14
2	酒石酸	149.1	73	-18
3	羟基乙酸	75.2	45.1	-16
3	羟基乙酸	75.2	47.1	-15
4	苹果酸	133.2	115	-16
4	苹果酸	133.2	71.2	-17
5	乳酸	89.2	43.1	-13
5	乳酸	89.2	45.1	-15
6	柠檬酸	191.1	111.1	-12
6	柠檬酸	191.1	87.1	-11
7	2-羟基丁酸	103.2	57.1	-13
7	2-羟基丁酸	103.2	45.1	-16
8	乳酸甲酯	104.9	45	+9
8	乳酸甲酯	104.9	33	+15
9	扁桃酸	151.2	107.1	-7
10	乳酸乙酯	118.9	91.1	+9
			45	+17
			47.1	+11
11	乳酸丁酯	147.3	45.1	+14
			91.1	+7
			63	+8
12	二苯乙醇酸	227.1	183.3	-9
13	柠檬酸三乙酯	299.2	210.7	+13
13	柠檬酸三乙酯	299.2	178.7	+15
14	羟基辛酸	159.2	113.2	-16
14	羟基辛酸	159.2	45	-17

组分	线性范围/（μg/mL）	回归方程	R²	检出限/（μg/g）	定量限/（μg/g）
葡糖醛酸	98.16~3 926	Y=0.151 7X-1.394 9	0.999 7	160	480
酒石酸	52.04~2 082	Y=0.656 3X+3.346 0	0.999 9	60	180
羟基乙酸	97.34~3 894	Y=0.196 3X-0.492 3	1	40	120
苹果酸	101.1~4 045	Y=0.341 6X-1.924 4	1	40	120
乳酸	123.6~4 942	Y=0.223 1X-3.759 0	1	100	300
柠檬酸	124.7~4 989	Y=0.329 2X-3.367 4	1	40	120
2-羟基丁酸	120.7~4 829	Y=0.173 2X-2.353 9	1	80	240
乳酸甲酯	122.1~4 885	Y=0.183 3X-0.167 7	1	80	240
扁桃酸	2.893~115.7	Y=9.793 5X+2.355 8	1	2	6
乳酸乙酯	100.3~4 013	Y=0.178 7X+0.883 2	1	80	240
乳酸丁酯	121.0~4 013	Y=0.182 8X-0.159 3	1	80	240
二苯乙醇酸	2.867~114.7	Y=17.024 1X-2.274 8	1	2	6
柠檬酸三乙酯	123.8~4 950	Y=17.024 1X-2.274 8	1	80	240
羟基辛酸	123.8~4 951	Y=0.146 4X+5.551 0	1	40	120

组分	低浓度回收率/%				RSD/%				高浓度回收率/%				RSD/%
组分	水	乳	凝胶	膏霜	水	乳	凝胶	膏霜	水	乳	凝胶	膏霜	水	乳	凝胶	膏霜
葡糖醛酸	113.3	107	103.7	95.8	2.7	1.7	0.8	0.6	99.2	102.9	101.1	91.6	0.4	0.4	0.7	0.8
酒石酸	94.7	98.7	97.1	90.4	0.5	0.9	1.8	1.5	100.3	101.5	100.8	106.2	0.3	0.6	0.8	0.7
羟基乙酸	93.4	103.2	96.4	97.1	0.8	0.5	1.8	2.1	98.5	102.5	100.2	101.9	0.4	0.6	0.8	0.5
苹果酸	102.1	106.1	104.6	102	0.2	0.6	0.7	1.7	98.2	101.6	100.3	102.6	0.6	0.7	0.7	0.6
乳酸	105.5	109.8	106.7	104.1	0.2	0.8	0.5	0.9	100.7	107.6	103.2	104.1	0.3	0.7	0.8	0.7
柠檬酸	104.2	107.3	106	103	0.3	0.4	0.8	1	99	101.5	100.4	102.4	0.4	0.6	0.8	0.6
2-羟基丁酸钠	105.4	107.1	96.9	103.1	3.1	0.7	0.8	0.8	98.7	101	98.2	102.3	0.7	0.7	0.9	0.5
乳酸甲酯	104.3	105.6	105.5	100.6	0.5	0.6	0.7	1	100.8	99.5	101	101.1	0.6	0.4	0.9	0.5
扁桃酸	106.1	109.2	107.3	95.6	0.8	0.6	1	1.1	99.3	102.1	99.3	101.4	0.6	0.7	1	0.5
乳酸乙酯	110.9	111.9	115.4	107.1	1.7	1.8	0.9	3.6	102	101	102.9	101.5	0.6	0.4	0.6	0.6
乳酸丁酯	109.9	112.8	112.9	104.8	0.6	0.6	0.9	2.4	103.5	100.2	102.9	98.3	0.5	0.7	0.6	0.9
二苯乙醇酸	87.8	98.2	98.3	97.7	1.9	1.8	3.2	1.9	97.4	88.4	91.8	87.7	0.3	2.3	2.2	2.1
柠檬酸三乙酯	100.5	109.3	103.4	103	0.4	2.8	1.4	0.7	99.1	102.2	99.7	99.4	0.4	0.9	0.9	0.6
羟基辛酸	99.5	101.5	100.7	99.3	4	0.5	3.8	1.3	98.6	91.4	96.8	92.5	0.4	1.4	1.2	1

检品名称	检出项	含量/%
××护理液	扁桃酸	0.99
××护理液	柠檬酸三乙酯	2.58
××皙白洁面乳	柠檬酸	0.21
××美白乳液	乳酸	0.08
××美白乳液	乳酸	0.08
××精华水	柠檬酸	0.32
××美白祛斑霜	乳酸	0.43
××美白亮肤爽肤水	柠檬酸	0.11

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