Shi Baoxian, Huang Weihua, Cheng Jieke
College of Chemistry and Molecular Science Wuhan University, Wuhan, PR China.
Electrophoresis. 2007 May;28(10):1595-600. doi: 10.1002/elps.200600615.
A sensitive fluorescence detection system with an Hg-lamp as the excitation source and a photon counter as the detector for microchip CE (MCE) has been developed. O-Phthaldialdehyde (OPA, lambda(ex) = 340 nm) was employed to label the catecholamine neurotransmitters such as dopamine (DA), norepinephrine (NE), and amino acid neurotransmitters including alanine (Ala), taurine (Tau), glycine (Gly), glutamic acid (Glu), and aspartic acid (Asp). The separation of seven derivatized neurotransmitters was successfully performed in MCE and the detection limits (S/N = 3) for DA, NE, Ala, Tau, Gly, Glu, and Asp were 0.85, 0.49, 0.23, 0.15, 0.13, 0.18, and 0.29 fmol, respectively. The system was then successfully applied for separation and determination of neurotransmitters in rat pheochromocytoma (PC 12) cells, and the average amounts of analyte per cell from a cell population were 2.5 fmol for DA, 3.3 fmol for Ala, 8.2 fmol for Tau, 4.0 fmol for Gly, and 1.9 fmol for Glu, respectively. By single-cell injection mode, electrophoresis separation and quantitative measurement of Glu in individual PC 12 cells was obtained. The average value of Glu per cell from single PC 12 cells analysis was found to be 3.5 +/- 3.1 fmol.
已开发出一种灵敏的荧光检测系统，该系统以汞灯作为激发源，以光子计数器作为微芯片毛细管电泳（MCE）的检测器。邻苯二甲醛（OPA，激发波长λ(ex)=340 nm）用于标记儿茶酚胺神经递质，如多巴胺（DA）、去甲肾上腺素（NE），以及氨基酸神经递质，包括丙氨酸（Ala）、牛磺酸（Tau）、甘氨酸（Gly）、谷氨酸（Glu）和天冬氨酸（Asp）。在MCE中成功实现了7种衍生化神经递质的分离，DA、NE、Ala、Tau、Gly、Glu和Asp的检测限（信噪比S/N = 3）分别为0.85、0.49、0.23、0.15、0.13、0.18和0.29 fmol。该系统随后成功应用于大鼠嗜铬细胞瘤（PC 12）细胞中神经递质的分离和测定，细胞群体中每个细胞的分析物平均含量分别为：DA 2.5 fmol、Ala 3.3 fmol、Tau 8.2 fmol、Gly 4.0 fmol和Glu 1.9 fmol。通过单细胞进样模式，实现了单个PC 12细胞中Glu的电泳分离和定量测量。单个PC 12细胞分析中每个细胞Glu的平均值为3.5±3.1 fmol。
