Department of Hygienic Analysis and Detection, The Key Laboratory of Modern Toxicology, Ministry of Education, School of Public Health, Nanjing Medical University, No. 101 Longmian Avenue, Nanjing 211166, China.
Department of Toxicology, Key Laboratory of Modern Toxicology, Ministry of Education, School of Public Health, Nanjing Medical University, No. 101 Longmian Avenue, Nanjing 211166, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Sep 1;1093-1094:91-99. doi: 10.1016/j.jchromb.2018.06.054. Epub 2018 Jun 25.
Tryptophan (TRP) and dopamine (DA) pathways are of great importance for their related pathology and physiology. In the present study, a new reliable and sensitive analytical method was developed and validated for 12 neuroactive metabolites in TRP and DA pathways in mouse serum and brain by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). The method exhibited good sensitivity as the lower limit of detections ranged from 0.10 to 0.50 ng/ml and the lower limit of quantifications ranged from 0.20 to 2.00 ng/ml by derivatization with dansyl chloride (DNS-Cl) following solid phase extraction (SPE) on C18 cartridges. Good linearity (R > 0.99), intra-day precision (<9.8% in serum and <8.8% in brain), inter-day precision (<8.9% in serum and <8.5% in brain) and accuracy (90.3%-110.3% in serum and 86.5%-114.0% in brain) were obtained. The method was successfully applied in measuring 12 neuroactive metabolites in TRP and DA pathways in serum and brain samples of male and female mice to explore the differences between genders. As a result, DA and the turnover of DA to 3,4-dihydroxyphenylacetic acid (DOPAC), 5-hydroxtryptamine (5-HT) to TRP and 5-hydroxyindole acetic acid (5-HIAA) to 5-HT in the serum and norepinephrine (NE) in the brain were significantly different between genders.
色氨酸（TRP）和多巴胺（DA）通路对于它们的相关病理和生理学非常重要。在本研究中，开发并验证了一种新的可靠且灵敏的超高效液相色谱-串联质谱（UHPLC-MS/MS）分析方法，用于检测小鼠血清和脑中 TRP 和 DA 通路中的 12 种神经活性代谢物。该方法通过衍生化后固相萃取（SPE），在 C18 小柱上用dansyl chloride（DNS-Cl）衍生化，具有良好的灵敏度，检测限范围为 0.10-0.50ng/ml，定量限范围为 0.20-2.00ng/ml。该方法具有良好的线性（R>0.99）、日内精密度（血清中<9.8%，脑内中<8.8%）、日间精密度（血清中<8.9%，脑内中<8.5%）和准确度（血清中 90.3%-110.3%，脑内中 86.5%-114.0%）。该方法成功应用于测量雄性和雌性小鼠血清和脑样本中 TRP 和 DA 通路中的 12 种神经活性代谢物，以探索性别之间的差异。结果表明，血清中的 DA 及其转化产物 3,4-二羟基苯乙酸（DOPAC）、5-羟色胺（5-HT）转化产物 TRP 和 5-羟吲哚乙酸（5-HIAA）转化产物 5-HT，以及脑内的去甲肾上腺素（NE）在性别之间存在显著差异。
