Probe sonication can pose challenges in small volumes due to potential overheating or foaming of the sample. Moreover, inconsistencies in sonication reproducibility may arise between samples or users. Furthermore, its limitation to processing one sample at a time restricts its applicability in high-throughput settings. Another limitation of this method is that extended sonication for 1 hour can lead to de-esterification of more than 5% of lipids. Additionally, titanium particles may detach and contaminate the solution.