I posted a few weeks ago but some new problems have come up and I could really use some advice! I am joining in on a research project in which the heavy metal content of mussels is being examined at 3 locations over 4 years. At each location, there would ideally be 4 sub-samples of around 10 mussels, for a total of 40 mussels. The 10 mussels in each sub-sample are then homogenized for heavy metal analysis. However, I just found out that in the last 2 years of the project, the mussels from each location have all been homogenized together and then divided into 4 replicates, rather than split up before homogenizing. This is now a case of "pseudoreplication," as there is technically only 1 sample with no variance for these 2 years.

What I am trying to find out now is: what would be the best way to analyze for spatial and temporal trends now that I technically only have 1 sample (40 mussels all homogenized into 1) for each location over the 4 years?

Thanks so much for taking the time.