UHPLC-MS/MS analysis of cAMP and cGMP in rat plasma as potential biomarkers of Yin-Yang disharmony in traditional Chinese medicine.

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.

UHPLC-MS/MS analysis of cAMP and cGMP in rat plasma as potential biomarkers of Yin-Yang disharmony in traditional Chinese medicine / 药物分析学报

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.

Further study of TCM for tumor / 中华中医药杂志

Based on the accurate cognition about tumor pathology,relying on observation of clinic cases,combining unsettled problems about tumor,instructed by theory of pathogenesis,the project puts forward the cancerous toxin-rooted and rising and falling of vital qi and pathogen-determined tumor viewpoint.The purpose of this paper is to provide methods and theory for treatment of tumor.