Liquid chromatography-mass spectrometry (LC-MS/MS) is a analytical technique which is used with High Performance liquid chromatography (HPLC) and the mass spectrometry. LC-MS/MS is commonly used technique in analytical laboratories for the qualitative and quantitative Determination of drug substances and biological samples. LC-MS has played a vital role in the evaluation and interpretation of bioavailability-bioequivalence and pharmacokinetic data. This review article reviews the recent Advancement in sample preparation, LC-MS/MS analysis of biological samples with LC-MS method development, validation, optimization, and different types of extraction procedures with different types of cartridges used in sample processing. In this review, we have described the Solid-phase extraction technique which is commonly used for sample preparation and reduces the matrix effect in LC-MS/MS analysis.