[Care and Use Manual 
]
Pico•Tag Amino Acid Analysis Column
I. Introduction
Waters exclusive bonding and packing processes, developed for the Pico•Tag® Column (3.9 mm x 15 cm), P/N 88131, produce a durable, high efficiency stationary phase ideally suited for the reverse-phase separation of phenylthiocarbamyl amino acids. Our stringent quality control procedures result in a column which, when used in conjunction with Waters Pico•Tag reagents, eluents, and HPLC system is guaranteed to provide the speed, resolution, and sensitivity needed for amino acid analysis.
Contents
I. introduction
II.PREPARATION FOR OPERATION
a.Column Installation
b.Solvent Requirements
c.Equilibration
This information provides the means to ensure that you derive the full benefit of our research, manufacturing, and quality control. For additional information on the complete method, refer to Section 4.4 of the Amino Acid Analysis Manual, P/N 07124, or the Pico•Tag Amino Acid Analysis System Operator’s Manual, P/N 88140.
WARNING: THE AMPULES OF PHENYLISOTHIOCYANATE WHICH ARE CONTAINED IN THE PICO•TAG CHEMICAL PACKAGE REACT ON CONTACT WITH STRONG ACIDS, EMITTING HIGHLY TOXIC CYANIDE FUMES AND/OR OXIDES OF SULPHUR. CARE MUST BE TAKEN TO ASSURE THAT THESE AMPULES AND CONTENTS ARE DISPOSED OF PROPERLY AND THAT THEY DO NOT COME INTO CONTACT WITH ACIDS.
Figure 1. Pico•Tag Method Amino Acid Analysis 250 pmol of Pierce H Standard
III.care and use
a.Sample and Eluent Preparation and Filtration
b.Precautions
c.Storage Considerations
d.Troubleshooting
e.Service and Applications Information
IV. column efficiency
V. TEST CONDITIONS
VI. RECOMMENDED SPARE PARTS (Steel Columns)
VII. WARRANTY
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[Care and Use Manual 
]
II. Preparation for operation
a. Column Installation
Remove the end plugs from your steel column with a 5/16” wrench and save them for storage after the column has been removed from the system. The column outlet is indicated by an arrow on the label (show-ing the direction solvent should flow). Tighten the fittings 1/4-to-1/2 turn beyond finger tight. DO NOT OVERTIGHTEN - THIS WILL DAMAGE THE FITTING SEAT. A properly prepared and assembled com-pression fitting in good condition is all that is required.
Follow the next four steps of this procedure if tubing cutting is required to connect a new column or to improve the end connections on your existing fittings.
1.Using a file with a cutting edge (such as the file included in the Startup Tool Kit, P/N 96146, supplied with each Waters Liquid Chromatography System) 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.
3.File the ends smooth and assemble as shown.
4.Slide the compression fitting, followed by the ferrule (large end of the taper first) over the tube. Be certain to bottom the tube in the fitting seat for which its use is intended to assure a leak-free connection.
Note: Attach a union in place of the column and flush the lines free of microparticulates before attaching the column.
Figure 5: Ferrule and Compression Assembly
b. Solvent Requirements
Your Pico•Tag column is shipped containing an aqueous/organic mixture compatible with the Pico•Tag eluents. It is recommended that only Waters Pico•Tag eluents be used with this column.
Waters Pico•Tag Eluent A (P/N 88108)
Waters Pico•Tag Eluent B (P/N 88112)
Waters Pico•Tag Diluent (P/N 88119)
c. Equilibration
A necessary step to successful use of your column is the initial solvation (or wetting) of the packing. Install the column in a column heater and bring temperature to 38 °C. Purge the column at 1 ml/min with Eluent B, then equilibrate with Eluent A for fifteen minutes. Equilibration between the mobile phase and packing is established when a stable baseline can be produced. If your result is un-satisfactory repeat the equilibration process.
III. care and use
Liquid chromatography columns have a finite life influenced by their care and use, number of injections, sample and solvent cleanliness, frequency of solvent changeover, and handling and storage procedures among other factors. If a change is observed in the:
Retention of a particular compound resolution between two compounds peak shape.
Take immediate steps to determine the reason for the changes, and until the determination is made, the results of any separations using the column must not be relied upon. Follow generally accepted pro-cedures for quality control and methods development when using these columns.
Important Note: Before running the first analysis on your new column perform the Test Sample Separation given in the Test Conditions Section.
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