Integrated LC-MS and network pharmacology methods to screen quantitative indicators in the Hippocampus histrix Kaup and method transfer

Hippocampus histrix Kaup is a popular marine medicine with high medicinal and healthcare values. In this study, liquid chromatography-mass spectrometry (LC-MS) analysis combined with network pharmacological method was used to screen for suitable quantitative indicators for the quality control of H. histrix Kaup. Firstly, an LC-MS analytical method for the simultaneous determination of 12 nucleosides in extracts of H. histrix Kaup was established. And then, a network pharmacological method incorporated target prediction, protein-protein interaction network, components-targets network, and targets-pathways network was performed to screen for quantitative indicators. Finally, the developed LC-MS method was transferred to liquid chromatographs to improve the generalizability of the method. All 12 nucleotides were authenticated in extracts of H. histrix Kaup by comparing with the standards. The optimal chromatographic separation conditions are as follows: the chromatographic separation was achieved on an Acquire BEH-C18 column (2.1 mm * 100 mm, 1.7 mu m) and gradient elution was performed using methanol solution and buffer (0.30% formic acid and 10 mmol/L ammonium acetate) as mobile phase at a flow rate of 0.15 mL/min and an acquisition wavelength of 260 nm. Network pharmacology results showed that adenosine, and uridine show excellent pharmacological activity. Integration the content, correlation, chromatographic separation, and pharmacological activity of each com- pound in H. histrix Kaup, uridine and adenosine were tentatively determined as quantitative indicators for quality control in H. histrix Kaup. The established LC-MS method was successfully transferred to liquid chromatographs, and the method is stable and reliable for the quality control of H. histrix Kaup. This developed integrated strategy was successfully used to screen quantitative indicators in the H. histrix Kaup.