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The “two-third spline” (2/3S) is a frequently applied method to detrend tree-ring series. It fits a spline with a 50% frequency cutoff at a frequency equal to two-thirds of each sample length in a dataset. It was introduced to ensure a minimum loss of low-frequency variance, which is resolvable during the detrending of ring-width series. In this paper I show potential problems that arise when rusing this method. The 2/3S runs counter the strengths of using a digital filter to detrend – i.e. one is giving up full control over the frequency-removing characteristics of the growth curve and each individual time series retains a different amount of low frequency. Thus, the 2/3S is less suitable for reconstructing climate or to compare environmental impacts on tree growth between groups – both of which comprise the majority of dendrochronological analyses – as it will likely introduce a temporal frequency bias. Within a long chronology it will result in decreasing power to resolve low frequencies towards present in a living-only trees setting, especially when the youngest segment lengths are 100 years and shorter, and more generally during the period where the chronology is constructed from samples with shorter segment lengths compared to the period with longer segment lengths. The frequency bias will also significantly impact regression slopes and correlation coefficients, possibly distorting analyses investigating multiple groups with different mean segment lengths. Highlighting these potential biases, I recommend the community to not use this method on an individual basis but rather to use a fixed spline stiffness for all samples based on the n% criterion (n = 67) of e.g. the mean segment length of the entire dataset.