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The waters that were present on early Mars may have been habitable. Characterising environments analogous to these waters and investigating the viability of their microbes under simulated martian chemical conditions is key to developing hypotheses on this habitability and potential biosignature formation. In this study, we examined the viability of microbes from the Anderton Brine Springs (United Kingdom) under simulated martian chemistries designed to simulate the chemical conditions of water that may have existed during the Hesperian. Associated changes in the fluid chemistries were also tested using inductively coupled plasma-optical emission spectroscopy (ICP-OES). The tested Hesperian fluid chemistries were shown to be habitable, supporting the growth of all of the Anderton Brine Spring isolates. However, inter and intra-generic variation was observed both in the ability of the isolates to tolerate more concentrated fluids and in their impact on the fluid chemistry. Therefore, whilst this study shows microbes from fluctuating brines can survive and grow in simulated martian water chemistry, further investigations are required to further define the potential habitability under past martian conditions.