Accurate and efficient amino acid analysis for protein quantification using hydrophilic interaction chromatography coupled tandem mass spectrometry

Table 1 Performance characteristics of 20 amino acids for quantitative analysis

	Retention time	^aRRF	^bLOD (pmol)	^cLOQ (pmol)	Min (pmol)^d	Max (pmol)^d
Alanine	5.90	0.59	11.62	35.21	0.50	1000
Arginine	7.41	42.39	0.83	2.53	0.10	1000
Asparagine	6.22	3.71	5.55	16.81	0.20	1000
Aspartate	6.69	4.66	7.76	23.52	0.30	1000
Cystine	7.38	5.83	0.20	0.61	0.15	750
Glutamine	6.18	1.39	0.04	0.12	0.03	150
Glutamate	6.39	9.42	0.44	1.32	0.10	1000
Glycine	6.11	0.34	8.42	25.53	2.50	1000
Histidine	7.22	51.16	1.18	3.58	0.10	1000
Isoleucine	4.95	9.87	0.14	0.42	0.10	1000
Leucine	4.83	57.03	1.73	5.23	0.10	1000
Lysine	7.58	24.58	2.55	7.72	0.50	1000
Methionine	5.08	10.77	0.10	0.30	0.10	1000
Phenylalanine	4.63	100.00	1.15	3.49	0.10	1000
Proline	5.40	96.77	0.78	2.36	0.10	1000
Serine	6.18	3.91	9.24	28.01	2.50	1000
Threonine	5.94	6.31	5.83	17.68	0.50	1000
Tryptophan	4.68	42.97	1.03	3.13	0.10	1000
Tyrosine	5.24	9.42	1.29	3.90	0.10	1000
Valine	5.33	28.45	0.49	1.48	0.10	1000

^aRRF represents response factors relative to phenylalanine which showed a maximum response. \({\text{RRF}} = 100*\left( {Peak\;Area\frac{analyte}{Phenylalanine}} \right)\)
^bLOD = 3.3 × σ/slope of the standard curve
^cLOQ = 10 × σ/slope of the standard curve
^d range of concentrations examined
(σ = SD of peak areas of least detectable concentration, n = 3)

ISSN: 1746-4811