Membrane proteins are proteins that are fully or partially embedded in the lipid manif important layer of the cell. They are biological macromolecules possessi cellular formation functions such as signaling, cell differentiation, cell protection, an cver current According to statistics, membrane proteins are the target of m drugs already present and they remain the main target in the development of group that is routinely drugs (Hopkins et al, 2002). In addition, this is also a protein used as a marker for disease diagnosis, cell differentiationassessment an well as delivery. Thus, this is the most important protein group for cellular activity as biological physiology and is of great interest Mass spectrometry is a core analytical tool in contemporary proteomics research, however, membrane protein sample preparation for mass spectrometric analysis has not been a routine task and generic protocols for this class of important proteins are rare. Removal of purposefully added interfering contaminants, mainly detergents, chaotropes, and salts, prior to Ms analysis is of vital importance, but nearly all existing protocols for achieving this goal are time-consuming, labor- intensive, or contribute to significant sample loss (Carpenter et al., 2008). A number of approaches have been developed to improve and facilitate the characterization of MP by MS (Arnold, 2007; Frick, 1993; Parish, 1986; Bunger, 2011). In this study, we focused on the use of the nanodiamond-based platform to enrich and prepare membrane protein samples from Hel a cell lines for shot-gun proteomics analyses. Especially, in mass spectrometry and membrane protein proteomics studies, nanodiamonds have several superior characteristics: (1) Nanodiamonds are inert and possess very strong chemical and physical properties that can withstand strong acidic and alkaline conditions. (2) The surface of nanodiamond after oxidation has a particularly high affinity for proteins or peptides so that nanodiamonds can capture and selectively recover these biological macromolecules without the need for other complex methods. Most notably, nanodiamonds originate from carbon, so their surface is hydrophobic. Thus, nanodiamonds have a higher affinity for membrane proteins