Amino Acids (9-Fluorenylmethyl-Chloroformate Derivatives) on Raptor™ ARC-18

PeakstR (min)Conc.
(µmole/mL)
1.Arginine (Arg)5.1450.4
2.Asparagine (Asn)5.5180.4
3.Glutamine (Gln)5.6300.4
4.trans-4-Hydroxyproline (Hyp)5.8780.4
5.Serine (Ser)6.1390.4
6.Aspartic acid (Asp)6.2560.4
7.Glutamic acid (Glu)6.6960.4
8.Threonine (Thr)7.0270.4
9.Glycine (Gly)7.4660.4
10.Alanine (Ala)8.2270.4
11.Proline (Pro)8.8030.4
12.Methionine (Met)9.3240.4
13.Valine (Val)9.5370.4
14.Norvaline9.6560.4
15.Tryptophan (Trp)9.9100.4
16.Phenylalanine (Phe)10.1200.4
17.Isoleucine (Ile)10.2140.4
18.Leucine (Leu)10.2850.4
19.Cystine10.7490.2
20.Histidine (His)12.0450.4
21.Lysine (Lys)12.1740.4
22.Cysteine (Cys)12.4630.4
23.Tyrosine (Tyr)12.8010.4
Amino acids are analyzed as 9-fluorenylmethyl-chloroformate derivatives.
image
LC_FF0539
ColumnRaptor™ ARC-18 (cat.# 9314A1E)
Dimensions:100 mm x 3 mm ID
Particle Size:2.7 µm
Temp.:30 °C
Sample
Diluent:0.1 N HCl
Conc.: 0.4 μmole/mL for each amino acid (0.2 μmole/mL for cystine)
Inj. Vol.:1 µL
Mobile Phase
A:0.1% Formic acid + 20 mM ammonium formate in water
B:0.1% Formic acid + 10 mM ammonium formate in 90:10 acetonitrile:water
Time (min)Flow (mL/min)%A%B
0.000.88020
6.250.86040
9.000.84060
10.000.84060
13.000.80100
13.010.88020
15.000.88020
DetectorUV/Vis @ 265, 4.8 nm
InstrumentWaters ACQUITY UPLC® H-Class
NotesDerivatization reaction: 50 μL amino acid mix + 100 μL 0.2 N borate buffer (pH 10.0) + 50 μL 15 mM 9-fluorenylmethyl-chloroformate solution + 50 μL acetonitrile

The injection can be performed after 5 minutes of reaction time.
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