Cyclic 3′,5′-adenosine monophosphate (cAMP) and cyclic 3′,5′-guanosine monophosphate (cGMP) are considered as potential biomarkers for Yin-Yang disharmony in traditional Chinese medicine. However, phosphodiesterase-mediated ex vivo degradation of these molecules in biological samples may result in their underestimation. In the present study, a ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for determination of cAMP and cGMP in rat plasma, with special consideration of their stability ex vivo. Following precipitation of proteins from plasma samples with 0.4 M perchloric acid, the analytes were chromatographed on a Shimadzu Shim-pack-XR-ODS II column with 2.5 mM ammonium acetate and methanol in gradient mode. The MS/MS detection was performed using multiple reaction monitoring in the positive electrospray ionization mode. The lower limit of quantification was 0.27 ng/mL for cAMP and 0.37 ng/mL for cGMP. The method was used to determine the plasma cAMP and cGMP levels in normal and Yin deficiency diabetic rats treated with or without Rehmannia glutinosa. The developed method may be useful for evaluating the regulatory effects of Chinese herbal medicine on the levels of cAMP and cGMP in the body.

环状 3',5'-腺苷酸 (cAMP) 和环状 3',5'-鸟苷酸 (cGMP) 被认为是中医阴阳失调的潜在生物标志物。然而，磷酸二酯酶介导的生物样品中这些分子的离体降解可能会导致它们被低估。在本研究中，开发了一种超高效液相色谱-串联质谱 (UHPLC-MS/MS) 方法来测定大鼠血浆中的 cAMP 和 cGMP，并特别考虑了它们的离体稳定性。在用 0.4 M 高氯酸从血浆样品中沉淀蛋白质后，分析物在 Shimadzu Shim-pack-XR-ODS II 柱上用 2.5 mM 醋酸铵和甲醇以梯度模式进行色谱分离。MS/MS 检测是在正电喷雾电离模式下使用多反应监测进行的。cAMP 的定量下限为 0.27 ng/mL，cGMP 的定量下限为 0.37 ng/mL。该方法用于测定正常和阴虚糖尿病大鼠的血浆cAMP和cGMP水平。地黄。所开发的方法可用于评估中草药对体内 cAMP 和 cGMP 水平的调节作用。