Capillary Separations Coupled to Electrospray MS:

Course Description:

This will be a one day course covering the current status of capillary separations (capillary LC, CE and CEC with column i.d. in the range of 25-500 um) coupled to electrospray MS and tandem MS and its capabilities to solve real world problems in proteomics, biochemistry, pharmaceutical, clinical and environmental analysis. The course covers the fundamentals of electrospray ionization with an emphasis on low flow electrospray (nanospray) and layer flow techniques for low flowrate operation with capillary separations. The criteria for selecting solvents and additives to achieve good capillary separations and compatibility with electrospray MS will be covered. Sample preparation methods to improve concentration detection limits will be reviewed. Operation tricks and tips to achieve reproducible and sensitive nanospray signals will be covered. The course will discuss the interpretation of electrospray mass spectra and CID mass spectra from tandem MS produced by peptides and substances of pharmaceutical and environmental significance. Additionally the advantages and disadvantages of capillary separations coupled with various mass analyzers (e.g. quadrupole, triple quadrupole, ion trap and time of flight MS) for qualitative and quantitative analysis will be reviewed. The course will conclude with a discussion on chip based separation-nanospray approaches for performing LC/MS and new technology available for on-line capillary MS. Problem sets given throughout the course will help reinforce the concepts covered.

Course Outline:

Introduction into capillary separations and electrospray mass spectrometry

Electrospray Mass Spectrometry

    · Theory of electrospray ionization.
    · Sample, solution and flowrate considerations.
    · Nano-electrospray- tricks and tips for obtaining a stable signals and good sensitivity.
    · Problem set reviewing material covered in this section.

Coupling capillary LC and capillary electrophoresis with electrospray MS

    · Interfaces - direct (nanospray) layer flow, and pneumatic assisted methods.
    · Plumbing and hardware to combine a capillary column to a nanospray tip.
    · Additive or electrolyte considerations for separations and MS.
    · Sample preparation (Extraction/Cleanup) designed for small sample and injection volumes.
    · Methods to improve detection limits.
    · What to do when incompatibilities exist between the capillary separation and electrospray MS.
    · Examples of capillary separations with MS analysis.
    · Problem set reviewing material covered in this section.

    Interpretation of Electrospray Spectra generated by LC/MS

    · High molecular weight compounds.
    · Low molecular weights compounds.
    · Generating CID mass spectra on triple quads, traps and Q-TOF instruments.
    · Basics of the interpretation of CID mass spectra.


    · Achieving the precision and accuracy with capillary separations combined to nanospray MS.
    · Stable isotope or homologue internal standards vs. external standards.
    · Problem set reviewing material covered in section IV and V.

    Future Trends/ Summary/Concluding Remarks

· Chip based separations combined with MS.
· Miniature MS systems.
· Capillary compatible EI and APCI interfaces for LC/MS.
· Summary and concluding remarks.

Student Profile:

  • This course is designed for the student who has completed an LC/MS short course or who has working knowledge of LC/MS and wants to improve their qualitative interpretation skills for problem solving in LC/MS.

Please direct all questions on this course offering to: