Waters Pico-Tag Columns for Free Amino Acids - Manual (Page 2)

[ Care and Use ManUal ]
The Pico•Tag Column for Free Amino Acids 2
Figure 1: Separations Method Using Pico•Tag Column For Free
Amino Acids II. InstallatIon
a. Solvent Requirements
Your Pico•Tag column is shipped containing an aqueous/organic mixture com-
patible with the Pico•Tag eluent. For best results use Pico•Tag Eluent 1 (P/N
10960) and Waters Pico•Tag Eluent 2 (P/N 10965) with this column.
b. Sample and Solvent Preparation and Filtration
• Use LC grade solvents, filtered to remove microparticulate matter above
0.45 µm. Pico•Tag prefiltered eluents eliminate the need for this step
by providing a prepared, ready-to-use solvent to reduce the problem of
plugged filters and preserve column life.
• Vacuum filtration or sonification may be used to remove dissolved
gasses which could affect your solvent delivery system.
• Waters Sample Clarification Kit, (P/N 26865) is recommended to filter
prepared samples and prevent excessive pressure buildup.
• Use of a Waters In-line Precolumn Filter (P/N 84560),is recommended.
• Pico•Tag Diluent, (P/N 88119), is recommended for reconstitution of
derivatized samples.
c. Column and Cartridge Installation
Remove the end plugs from your column with a 5/16” wrench. (Be sure
to replace the end plugs when the column is removed from the system for
storage.) The column outlet is indicated by an arrow on the label show-
ing the direction solvent should flow. Tighten the fittings to turn. DO NOT
OVERTIGHTEN - THIS WILL DAMAGE THE FITTING SEAT. A properly prepared and
assembled compression fitting in good condition is all that is required.
Follow the next three steps of this procedure if you must cut tubing to con-
nect a new steel column or to improve the end connections on your existing
fittings.
1. Using a three-cornered file with a cutting edge scribe the circumference
of the tubing at the desired break.
2. Grasp the tubing on both sides of the scribe mark with cloth-covered
pliers (to prevent marring the tube surface) and gently work the tube
back and forth until it separates.
Abbreviation Component Name Retention Time
PSER Phosphoserine 3.05
ASP Aspartic Acid 3.50
GLU* Glutamic Acid 4.01
AAD α-Aminoadipic Acid 5.62
HYPRO Hydroxyproline 7.12
PEA Phosphoethanolamine 7.53
SER* Serine 9.20
ASN* Asparagine 9.50
GLY* Glycine 10.30
_-ALA β Aline 12.62
TAU Taurine 14.82
HIS* Histidine 17.00
GABA γ-Aminobutyric Acid 18.00
CIT Citrulline 18.62
NH2 Ammonia 19.62
THR Threonine 20.35
ALA Alanine 21.52
BAIB B-Aminoisobutyric Acid 23.65
CARN Carnosine 24.72
ARG* Arginine 26.02
METSO2 Methionine Sulfone 27.00
PRO Proline 30.15
1MH 1-Methyl Histidine 31.32
3MH 3-Methyl Histidine 32.25
AAB A-Aminobutyric Acid 37.99
TYR* Tyrosine 44.89
VAL* Valine 47.62
MAT* Methionine 49.22
CYST Cystathionine 51.35
RG1 Reagent 1 52.90
CYS2* Cystine 52.99
RG2 Reagent 2 54.40
ILE Isoleucine 54.95
LEU* Leucine 55.65
HYLYS 1 Hydroxylsine 1 58.15
HYLYS 2 Hydroxylsine 2 58.90
PHE* Phenylanlanine 60.02
TRP* Tyrptophan 60.32
ORN Orninthine 60.77
LYS* Lysine 65.52