a 大连理工大学 制药科学与技术学院 大连 116024;
b 大连化学物理研究所 生物技术部 大连 116023;
c 大连医科大学 口腔医学院 大连 116044
Ultrasensitive Detection of Hair Cortisol Based on Portable Raman Spectrometer and Double-layer Paper Microdevice
Gao Zhiganga, Zheng Tingtinga, Deng Jiua, Li Xiaoruia, Qu Yueyanga, Lu Yaob, Liu Tingjiaoc, Luo Yonga, Zhao Weijiea, Lin Bingchenga
a School of Pharmaceutical Science and Technology, Dalian University of Technology, Dalian 116024;
b Biological Division, Dalian Institute of Chemical Physics, Dalian 116023;
c College of Stomatology, Dalian Medical University, Dalian 116044
Cortisol in the human hair is a clinical biomarker of long-term social-and-work-related mental stress, which is of high morbidity rate in the current modern society. This study developed a sensitive hair cortisol assay, featuring sur-face-enhanced Raman spectroscopy, immunoreaction, and a double-layer paper microdevice. The first layer of the paper mi-crodevice was used to remove the hair residue in the hair extract by filtration (sample pretreatment). The second layer was used for competitive immunoreaction and detection. Standard cortisol antigen immobilized in the second layer and the free cortisol in hair extract competed to bind the spiked Raman-active cortisol monoclonal antibody solution. The hair cortisol can be quantitated by the intensity of Raman signal of monoclonal antibody bound on the paper. We found the Raman signal decreased as the cortisol concentration increases in hair samples. The relative Raman intensity measured was linearly proportional to the logarithmic value of the cortisol concentration in hair samples we measured. The detection of limit (LOD) was 1 pg/mL with the portable Raman spectrometer. The RSD of measurement was 8.38% (n=6). In addition, we used LC-MS to measure two real samples as a comparison with our method as above. The results are 0.771 and 0.153 ng/mL by LC-MS method and 0.63 and 0.247 ng/mL by the proposed method. It can be observed that the results are in same order, demonstrating the validity of the proposed method. In addition, 48 samples can be measured in a single chip. These results showed that this method is sensitive, specific, and suitable for large-scale screening of hair cortisol samples.
高志刚, 郑婷婷, 邓九, 李晓瑞, 曲玥阳, 陆瑶, 刘婷娇, 罗勇, 赵伟杰, 林炳承. 基于便携拉曼光谱仪和双层纸芯片的头发皮质醇超高灵敏检测[J]. 化学学报, 2017, 75(4): 355-359.
Gao Zhigang, Zheng Tingting, Deng Jiu, Li Xiaorui, Qu Yueyang, Lu Yao, Liu Tingjiao, Luo Yong, Zhao Weijie, Lin Bingcheng. Ultrasensitive Detection of Hair Cortisol Based on Portable Raman Spectrometer and Double-layer Paper Microdevice. Acta Chim. Sinica, 2017, 75(4): 355-359.