Amino Acids in Human Plasma on Raptor Polar X

	Peaks	t_R (min)	Precursor Ion	Product Ion
1.	Ethanolamine	1.06	62.0	44.1
2.	Acetyltyrosine	1.83	224.1	136.1
3.	γ-Aminobutyric acid	2.25	104.0	69.0
4.	β-Aminoisobutyric acid	2.27	104.1	30.0
5.	β-Alanine	2.52	90.0	30.1
6.	Tryptophan	3.31	205.1	146.1
7.	Leucine	3.40	132.1	86.1
8.	Phenylalanine	3.53	166.1	120.1
9.	Isoleucine	3.60	132.1	86.1
10.	Alloisoleucine	3.77	132.1	86.1
11.	Methionine	4.15	150.1	104.1
12.	Valine	4.27	118.1	72.1
13.	Tyrosine	4.27	182.1	136.1
14.	Arginine	4.52	175.2	70.1
15.	α-Aminobutyric acid	4.57	104.0	58.1
16.	Pipecolic acid	4.60	130.0	84.1
17.	1-Methylhistidine	4.62	170.1	124.1
18.	Taurine	4.65	126.1	108.1
19.	Histidine	4.65	156.1	110.2
20.	Anserine	4.67	241.2	109.1
21.	Carnosine	4.67	227.2	110.1
22.	3-Methylhistidine	4.68	170.0	126.0

	Peaks	t_R (min)	Precursor Ion	Product Ion
23.	Lysine	4.68	147.1	84.1
24.	Ornithine	4.72	133.1	70.0
25.	Hydroxylysine	4.84	163.1	128.1
26.	Alanine	4.86	90.0	44.1
27.	Proline	4.89	116.1	70.1
28.	Sarcosine	5.05	90.0	44.1
29.	Threonine	5.15	120.1	74.1
30.	Glycine	5.19	76.1	30.1
31.	Hydroxyproline	5.26	132.1	86.1
32.	Adenosylhomocysteine	5.30	385.0	133.7
33.	Glutamine	5.43	147.1	84.1
34.	Homocitrulline	5.43	190.1	84.1
35.	Serine	5.46	106.1	60.1
36.	α-Aminoadipic acid	5.53	162.1	98.1
37.	Asparagine	5.54	133.1	74.1
38.	Citrulline	5.61	176.1	159.1
39.	Homocystine	6.20	269.1	136.0
40.	Glutamic acid	6.26	148.1	84.1
41.	Cystathionine	6.75	223.1	134.0
42.	Cystine	6.99	241.1	152.0
43.	Phosphoethanolamine	7.32	142.0	44.1
44.	Argininosuccinic acid	7.41	291.1	70.1
45.	Aspartic acid	8.00	134.1	74.1

Column

Raptor Polar X (cat.# 9311A12)

Dimensions:

100 mm x 2.1 mm ID

Particle Size:

2.7 µm

Pore Size:

90 Å

Guard Column:

Raptor Polar X EXP guard column cartridge 5 mm, 2.1 mm ID, 2.7 µm (cat.# 9311A0252)

Temp.:

35 °C

Standard/Sample

Conc.:

Endogenous Levels (~μmol/L)

Inj. Vol.:

5 µL

Mobile Phase

Water, 0.5% formic acid, 1 mM ammonium formate

90:10 Acetonitrile:water, 0.5% formic acid, 1 mM ammonium formate

Time (min)	Flow (mL/min)	%A	%B
0.00	0.3	4	96
2.00	0.3	4	96
10.00	0.3	70	30
10.01	0.3	95	5
11.00	0.3	95	5
11.01	0.3	4	96
13.00	0.3	4	96

Detector	MS/MS
Ion Mode:	ESI+
Mode:	MRM
Instrument	UHPLC
Sample Preparation	An aliquot of 50 µL of plasma (Control Plasma Level II, Chromsystems) was mixed with 5 µL of 30% sulfosalicylic acid solution for protein precipitation. Following centrifugation at 4200 rpm for 10 minutes, a 27.5 µL aliquot of clear supernatant was mixed with 2 µL of internal standard working solution and 225 µL of mobile phase B prior to LC-MS/MS analysis.

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