• Excellent Instrumentation is available

• Quantitative analysis of 16 amino acids

• Variable susceptibility to acid hydrolysis
  Trp, Cys - destroyed
  Ser, Thr - partially destroyed
  Ile, Val - slow to cleave
  Met - subject to oxidation
  Asn, Gln - deamidated

• Requires
  technical and mechanical skills
  a quantitative mind
  common sense
  rigorous system maintenance.

• Dedicated instrumentation is ideal.

Amino Acid Analysis - Applications

• Quantify peptides and proteins
• Corroborate synthetic peptide structures
• Corroborate recombinant protein structures
• Identify proteins based on AA composition
• Design fragmentation strategies
• Detect odd amino acids

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Complications in Amino Acid Analysis

• Inherent Amino Acid Variability
• Sample Preparation & Background
• Instrument Performance & Maintenance
• Data Reduction/Interpretation
• Operator Performance and Work Habits

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Amino Acid Analysis

Methods and instrumentation used to analyze ABRF-93 AAA

Methods

Instrumentation

Common Amino Acids

Common Amino Acids

Phosphoamino Acids

Pre-column	32	Pre-column	32	Pre-column	25
PTC	29	ABI 420	2	----	2
OPA/FMOC	2	ABI 420A	10	----	7
AQC*	1	ABI 420H	2	----	3
Post-column	18	Beckman 110B	1	Beckman 126	1
ninhydrin	15	Beckman 421A	1	Beckman PACE Ý	1
OPA	3	HP 1090 L	1	----	1
		HP 1090 M	1	----
Phosphoamino Acids		HP 1090 AQII	2	----	2
Pre-column	25	Waters Pico-Tag	9	----	5
PTC	19	Waters 6000	1	----	1
OPA/FMOC	2	Waters 625	1	----	1
AQC*	1	Waters	1	----	1
DABSYL	3	Post-column	17	Post-column	11
		Beckman 6300	14	----	7
Post-column	11	ISCO 2360	1	----	3
ninhydrin	8	JEOL 300	1	----	2
OPA	3	Waters	1	----	1

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*AQC: 6 aminoquinolyl-N-hydroxysuccinimidyl carbamate

+Capillary zone electrophoresis

Yuksel et. al., (1994) in Techniques in Protein Chemistry V (J.W.Crabb, ed.), Academic Press, pp. 231-240.

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