Spike sorting, an indispensable process in the analysis of neural biosignals, aims to segregate individual action potentials from mixed recordings. This study delves into a comprehensive investigation of diverse unsupervised classification algorithms, some of which, to the best of our knowledge, have not previously been used for spike sorting. The methods encompass Principal Component Analysis (PCA), K-means, Self-Organizing Maps (SOMs), and hierarchical clustering. The research draws insights from both macaque monkey and human pancreatic signals, providing a holistic evaluation across species. Our research has focused on the utilization of the aforementioned methods for the sorting of 327 detected spikes within an in vivo signal of a macaque monkey, as well as 386 detected spikes within an in vitro signal of a human pancreas. This classification process was carried out by extracting statistical features from these spikes. We initiated our analysis with K-means, employing both unmodified and normalized versions of the features. To enhance the performance of this algorithm, we also employed Principal Component Analysis (PCA) to reduce the dimensionality of the data, thereby leading to more distinct groupings as identified by the K-means algorithm. Furthermore, two additional techniques, namely hierarchical clustering and Self-Organizing Maps, have also undergone exploration and have demonstrated favorable outcomes for both signal types. Across all scenarios, a consistent observation emerged: the identification of six distinctive groups of spikes, each characterized by distinct shapes, within both signal sets. In this regard, we meticulously present and thoroughly analyze the experimental outcomes yielded by each of the employed algorithms. This comprehensive presentation and discussion encapsulate the nuances, patterns, and insights uncovered by these algorithms across our data. By delving into the specifics of these results, we aim to provide a nuanced understanding of the efficacy and performance of each algorithm in the context of spike sorting.