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1. A chromatographic separation device for preparing a sample for proteomics analysis, comprising: (a) a sample preparation device which comprises: (1) a pipette tip of about 200 uL; (2) a solid-phase extraction membrane with a length of about 3 mm filled at the bottom end of the pipette tip; and (3) a layer of strong cation exchange resin or strong anion exchange resin located above the solid-phase extraction membrane, wherein the sample preparation device does not comprise a second solid-phase extraction membrane located above the layer of strong cation exchange resin or strong anion exchange resin; (b) a lysis buffer for lysing a biological sample into a plurality of proteins, comprising HEPES, NaCl, guanidine hydrochloride, n-dodecyl β-D-maltoside (DDM), and a protease inhibitor; (c) a digesting reagent for digesting the plurality of proteins retained in the layer of strong cation exchange resin or strong anion exchange resin into a plurality of peptides, comprising trypsin; (d) a transfer buffer for transferring the plurality of peptides in the layer of strong cation exchange resin or strong anion exchange resin into the solid-phase extraction membrane, comprising sodium chloride, ammonium formate or ammonium bicarbonate; and (e) an elution buffer for eluting the plurality of peptides from the solid-phase extraction membrane, comprising (1) ammonium formate or acetic acid, and (2) acetonitrile.
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