This study presented for the first time the development and validation of a sensitive method for quantification of dopamine, noradrenaline, and adrenaline in Krebs–Henseleit solution by LC–tandem mass spectrometry. Aliquots of 2.0 mL calibrators, quality controls, and samples of Krebs–Henseleit solution incubated with tortoise's aortic ring for 30 min were extracted by solid‐phase extraction. Catecholamine separation was achieved on a 100 × 4.6 mm LiChrospher RP‐8 column and the quantification was performed by a mass spectrometer equipped with an electrospray interface operating in positive ion mode. The run time was 4 min and the calibration curve was linear over the range of 0.1–20.0 ng/mL. The method was applied to the measurement of basal release of dopamine, noradrenaline, and adrenaline from the tortoise Chelonoidis carbonaria aortae in vitro. One aortic ring (30 mm) per tortoise (n = 5) was incubated for 30 min in a 5 mL organ bath filled with Krebs–Henseleit solution. The method demonstrated sensitivity, precision, and accuracy enough for its application in the measurement of basal release of these catecholamines from C. carbonaria aortic rings in vitro. The mean (standard deviation) concentrations of dopamine, noradrenaline, and adrenaline were 3.48 (2.55) ng/mL, 1.40 (0.57) ng/mL, and 1.87 (1.09) ng/mL, respectively.

中文翻译：

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液相色谱-串联质谱法测定克雷伯斯-亨塞利特溶液中多巴胺，去甲肾上腺素和肾上腺素的含量，并测定离体螯虾的基本释放量

这项研究首次提出了通过LC串联质谱法定量分析Krebs-Henseleit溶液中多巴胺，去甲肾上腺素和肾上腺素的灵敏方法的开发和验证。通过固相萃取法提取了2.0 mL校准物，质量控制样品以及与乌龟主动脉环孵育30分钟的Krebs-Henseleit溶液样品。儿茶酚胺的分离是在100×4.6 mm LiChrospher RP-8色谱柱上进行的，定量是通过配有以正离子模式运行的电喷雾接口的质谱仪进行的。运行时间为4分钟，校准曲线在0.1–20.0 ng / mL范围内呈线性。该方法用于测量乌龟中多巴胺，去甲肾上腺素和肾上腺素的基础释放体外螯虾石carbon 。每个乌龟（n  = 5）一个主动脉环（30毫米）在装有Krebs-Henseleit溶液的5毫升器官浴中孵育30分钟。该方法显示出足够的灵敏性，精密度和准确性，可用于测量体外从碳弧菌主动脉环中这些儿茶酚胺的基础释放。多巴胺，去甲肾上腺素和肾上腺素的平均浓度（标准差）分别为3.48（2.55）ng / mL，1.40（0.57）ng / mL和1.87（1.09）ng / mL。