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

Amino acids are analyzed as 9-fluorenylmethyl-chloroformate derivatives.

Column

Raptor ARC-18 (cat.# 9314A1E)

Dimensions:

100 mm x 3 mm ID

Particle Size:

2.7 µm

Temp.:

30 °C

Standard/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

0.1% Formic acid + 20 mM ammonium formate in water

0.1% Formic acid + 10 mM ammonium formate in 90:10 acetonitrile:water

Time (min)	Flow (mL/min)	%A	%B
0.00	0.8	80	20
6.25	0.8	60	40
9.00	0.8	40	60
10.00	0.8	40	60
13.00	0.8	0	100
13.01	0.8	80	20
15.00	0.8	80	20

Detector	UV/Vis @ 265, 4.8 nm
Instrument	Waters ACQUITY UPLC H-Class
Sample Preparation	Derivatization 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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