Amino Acid Analysis Course - 2 days

Course duration:

Laserchrom, Rochester.

Summary:

The course covers amino acid analysis using HPLC and with an analyser. By the end of the course, the analyst will know the merits of the different detection methods and derivatisation reagents, the relative merits of reversed phase and ion exchange, and when to use HPLC and when to use an analyser.

Course content:

Introduction to amino acids.

• Those used in feedstuffs
• Those found in protein hydrolysis
• Thoses found in physiological fluids

Use of reversed phase for amino acid separation

• Eluents for use with reversed phase
• Gradient profiles
• Limitations of reversed phase

Use of ion-exchange for amino acid separation

• Eluents for use with ion exchange
• Gradient profiles
• Limitations of ion exchange

Reasons why retention times vary in HPLC methods

Using UV, Visible or Fluorescence detection

• Sensitivity
• Robustness
• Selectivity

Derivatisation reagents

• OPA
• Ninhydrin
• Phenythiohydantoin (PTH)
• Phenylisothiocyanate (PITC)
• Dansyl chloride
• FMOC

Maximum peak numbers with HPLC methods

Amino acid analysis using an analyser

Comparison of stepwise solvent changes and gradient elution

Setting up the gradient profile

Column types for amino acid analysers

Day 2 is a practical day, running the samples in the lab.

Course Dates in 2010