[Microchip-based reversed-phase liquid chromatography-tandem mass spectrometry platform for protein analysis]

Area of Science:

• Analytical Chemistry
• Biochemistry

Context:

• Microchip-based separations are gaining attention for high throughput and sensitivity.
• Hyphenation of microchip liquid chromatography with tandem mass spectrometry (MS/MS) is crucial for complex biological sample analysis.

Purpose:

• To establish and evaluate a microchip-based reversed-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS) platform for protein identification.
• To demonstrate the platform's sensitivity, reproducibility, and potential for rapid protein analysis.

Summary:

• A novel RPLC-MS/MS platform was developed using monolithic materials in microchannels as trap and separation columns.
• Conventional micro-liquid chromatography components and a chip-MS interface were integrated for fluidic control.
• The platform successfully analyzed 100 ng of digested bovine serum albumin (BSA), achieving high sequence coverage (34.10-39.37%) with good reproducibility (RSD of 7.3%).
• Chip-to-chip reproducibility was confirmed by analyzing BSA using microchips from different batches, yielding comparable results.

Impact:

• The established platform demonstrates high sensitivity and good reproducibility for protein analysis.
• This technology shows great potential for rapid and efficient protein identification in complex biological samples.
• The findings contribute to advancements in microfluidic analytical devices for proteomics.