Validated UPLC-MS/MS quantification and intracellular PK-PD Modeling of periplocin-related cardiac glycosides in H/R-injured H9c2 cells.

Peng Zhao; Fanjiao Zuo; Caixia Li; Yanjin Li; Haoran Wu; Yingjing Zhao; Yameng Zhu; Jun He

doi:10.1016/j.jchromb.2026.125227

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Validated UPLC-MS/MS quantification and intracellular PK-PD Modeling of periplocin-related cardiac glycosides in H/R-injured H9c2 cells.

Peng Zhao·Fanjiao Zuo·Caixia Li·Yanjin Li·Haoran Wu·Yingjing Zhao·Yameng Zhu·Jun He

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Abstract

Reliable intracellular quantification is essential for characterizing the target-site disposition and exposure-response relationships of bioactive natural products. In this study, an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the simultaneous determination of periplocin and four related cardiac glycoside metabolites in H9c2 cell lysates. Acceptable linearity, precision, recovery, and stability were achieved for intracellular quantification. Cells were treated with each compound at 50 μM, and intracellular concentrations and cell viability were monitored over 48 h. In hypoxia/reoxygenation (H/R) -injured cells, the time to maximum intracellular concentration was shortened for all five compounds, indicating altered cellular disposition under injury conditions. Cell viability was improved by all compounds during the observation period. Pharmacokinetic-pharmacodynamic (PK-PD) integration was performed using a sigmoid Emax model, and acceptable model fits were obtained, with Akaike information criterion (AIC) values ranging from 79.22 to 130.46. Low apparent EC50 values were estimated under this single-dose design, whereas the estimated Ke0 values suggested delayed equilibration with the effect compartment. These findings indicate that sustained cytoprotective responses can be produced by periplocin and related metabolic markers in injured cardiomyocytes. This intracellular bioanalytical strategy provides a quantitative approach for linking cellular exposure to pharmacodynamic response and may support further evaluation of periplocin-related cardiac glycosides.

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Tandem Mass Spectrometry Animals Cardiac Glycosides Liquid Chromatography-Mass Spectrometry Chromatography, High Pressure Liquid Reproducibility of Results Rats Cell Line Linear Models Cell Survival Saponins Myocytes, Cardiac Limit of Detection Cardiac glycosides Intracellular quantification PK-PD modeling Periplocin UPLC-MS/MS Journal Article

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Peng Zhao, Fanjiao Zuo, Caixia Li, Yanjin Li, Haoran Wu, Yingjing Zhao, Yameng Zhu, Jun He. 2026-07-29. Validated UPLC-MS/MS quantification and intracellular PK-PD Modeling of periplocin-related cardiac glycosides in H/R-injured H9c2 cells.. https://doi.org/10.1016/j.jchromb.2026.125227

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