Bao De-Jun, Feng Zhuang-Zhuang, Zhang Xu, Sun Qi, Zhang Zhuo-Na, Hu Xiao-Jian, Zhu Ying, Lin Xiao
China CDC Key Laboratory of Environment and Population Health,National Institute of Environmental Health,Chinese Center for Disease Control and Prevention,Beijing 100021,China.
Se Pu. 2025 Jun;43(6):620-629. doi: 10.3724/SP.J.1123.2024.10031.
In this study, a method for the simultaneous determination of 22 organic ultraviolet absorbers (OUVs) in human serum was established by combining protein precipitation technology (PPT), efficiency lipid removal technology (ELR) and ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The OUVs include five benzophenone compounds, five benzotriazole compounds, two cinnamate ester compounds and their three metabolites, two salicylate compounds, two camphor derivative compounds, one triazine compound, one dibenzoylmethane compound and one amino aminobenzoic acid derivative compound. Chromatography was performed using an Acquity BEH C column (100 mm×2.1 mm, 1.7 μm), gradient elution was carried out using methanol-water and methanol-0.1% ammonia water as the mobile phases. Compounds were detected in both positive and negative electrospray ionization (ESI/ESI) modes using multiple reaction monitoring (MRM), and quantified using stable-isotope internal standards. The experimental results showed that the 22 target compounds exhibited good linear relationships within their respective linear ranges, with the correlation coefficients ()≥0.999 3. The method detection limits (MDLs) ranged from 0.02 to 0.48 ng/mL, and the method quantification limits (MQLs) ranged from 0.02 to 1.60 ng/mL. At the three spiked levels of low, medium and high, the spiked recoveries of the 22 target analytes ranged from 79.9% to 136.1%, the intra-day precisions were from 1.5% to 25.4%, and the inter-day precisions were from 0.6% to 23.5%. After correction by the stable-isotope internal standard method, the matrix effects of the 22 target analytes in fetal bovine serum were 83.0%‒119.9%. The developed method was successfully used to detect 22 OUVs in 110 human serum samples. With the exception of 3-benzylidene camphor (3-BC), 2-(2-benzotriazol-2-yl)-4-methyl-6-(2-propenyl)phenol (UV-9), 4-methylbenzylidene camphor (4-MBC), 2,2'-dihydroxy-4-methoxybenzophenone (BP-8), and 2,4-dihydroxybenzophenone (BP-1), which were not detected, the remaining 17 substances were detected with overall detection rates of 0.9%-65.5% and the detection levels were <MQL‒11.7 ng/mL. The developed analytical method is simple, convenient, highly sensitive, and is expected to become an effective tool for quantifying 22 OUVs in human serum.
本研究通过结合蛋白沉淀技术(PPT)、高效脂质去除技术(ELR)和超高效液相色谱 - 串联质谱法(UPLC-MS/MS),建立了一种同时测定人血清中22种有机紫外线吸收剂(OUVs)的方法。这些OUVs包括5种二苯甲酮化合物、5种苯并三唑化合物、2种肉桂酸酯化合物及其3种代谢物、2种水杨酸酯化合物、2种樟脑衍生物化合物、1种三嗪化合物、1种二苯甲酰甲烷化合物和1种氨基氨基苯甲酸衍生物化合物。采用Acquity BEH C柱(100 mm×2.1 mm,1.7 μm)进行色谱分析,以甲醇 - 水和甲醇 - 0.1%氨水为流动相进行梯度洗脱。采用多反应监测(MRM)在正、负离子电喷雾电离(ESI/ESI)模式下对化合物进行检测,并使用稳定同位素内标进行定量。实验结果表明,22种目标化合物在各自线性范围内呈现良好的线性关系,相关系数(r)≥0.999 3。方法检出限(MDLs)为0.02~0.48 ng/mL,方法定量限(MQLs)为0.02~1.60 ng/mL。在低、中、高3个加标水平下,22种目标分析物的加标回收率为79.9%~136.1%,日内精密度为1.5%~25.4%,日间精密度为0.6%~23.5%。经稳定同位素内标法校正后,22种目标分析物在胎牛血清中的基质效应为83.0%‒119.9%。所建立的方法成功用于检测110份人血清样品中的22种OUVs。除未检测到3 - 亚苄基樟脑(3 - BC)、2 - (2 - 苯并三唑 - 2 - 基) - 4 - 甲基 - 6 - (2 - 丙烯基)苯酚(UV - 9)、4 - 甲基亚苄基樟脑(4 - MBC)、2,2'-二羟基 - 4 - 甲氧基二苯甲酮(BP - 8)和2,4 - 二羟基二苯甲酮(BP - 1)外,其余17种物质均被检测到,总体检出率为0.9% - 65.5%,检测水平为<MQL‒11.7 ng/mL。所建立的分析方法简便、灵敏,有望成为定量人血清中22种OUVs的有效工具。