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CORTECS 2.7 M Columns: Cont Ents
CORTECS 2.7 M Columns: Cont Ents
7 µm Columns
II. G E T T ING STA RT E D c. Minimizing Band Spread Volume
Each CORTECS 2.7 µm Column comes with a Certificate of Band spreading is the measurement of the system dispersion,
Analysis, which includes the bonded phase batch number and which impacts the chromatographic performance. Internal tubing
the analytical test results for the unbonded and bonded particle. diameter and fluidic connections can significantly impact system
T he included Performance Test Chromatogram summarizes band spreading and chromatographic performance. Larger
the performance of each individual column and provides batch tubing diameters cause excessive peak broadening and reduced
number, column serial number, USP plate count, USP tailing sensitivity (Figure 1).
factor, retention factor, and chromatographic test conditions.
These data should be recorded and stored for future reference. 0.005 inches
When available, the information can be accessed via the
ACQUITY UPLC Console using the attached eCord. 0.020 inches
0.040 inches
a. Column Connection
CORTECS 2.7 µm Columns are designed to operate on any HPLC,
UHPLC, or UPLC system. Due to the absence of an industry
standard, be aware that the type of fittings and connection on Diluted/distorted sample band
2. Purge the pumping system to 100% organic mobile phase Note. If mobile phase additives (i.e., ion-pairing reagents) are
(methanol or acetonitrile). present in low concentrations (<0.2% v/v), 100 to 200 column
3. Connect the inlet of the column to the chromatographic system. volumes may be required for complete equilibration. In addition,
4. Flush the column with 100% organic mobile phase (methanol mobile phases that contain formate (i.e., ammonium formate,
or acetonitrile) by setting the pump flow rate to 0.1 mL/min. formic acid) may require extended equilibration times.
Increase the pump flow rate to 0.5 mL/min over 5 minutes.
For reversed-phase separations, equilibrate the column with a
5. Once the mobile phase is flowing from the column outlet, stop
minimum of 10 column volumes of the mobile phase to be used
the flow.
(refer to Table 1 for a list of column volumes). The column may be
6. Attach the column outlet to the detector. This prevents air
considered fully equilibrated once a constant backpressure and a
from entering the detector flow cell.
stable detector baseline is achieved.
7. Increase the flow rate as described in step 4.
VanGuard Cartridge Columns require the VanGuard Cartridge Holder (p/n 186007949) for proper assembly.
/ " wrench
7 16
/ " wrench
3 8
O-Ring Compression Screw Transition Tube Assembly
ATTENTION: READ AND 1. Remove protective o-ring. 2. T hread the holder assembly 3. With holder on top, tighten holder
finger tight into the to the column inlet using wrenches
UNDERSTAND ALL ASSEMBLY Keep assembly upright to
keep ferrule from falling off. column inlet. (1/4 turn after finger tight).
INSTRUCTIONS PRIOR TO USE.
Hold assembly securely
DO NOT remove the protective Two Piece Ferrule to prevent the holder Ensure that the
plug from the holder assembly from slipping from holder is firmly
the column inlet. seated into the
until Step 4. T he plug is used to column inlet before
seat the tubing securely prior to setting the ferrule.
final assembly. HPLC column
⁄ " wrench
38
Guard cartridge
1. Loosen guard cartridge 2. Insert new guard cartridge 3. Tighten guard cartridge.
and remove from holder. into holder. 7 16
⁄ " wrench
The guard cartridge can be replaced
without removing the holder from
the column inlet.
⁄ " wrench
7 16
⁄ " wrench
38
38
⁄ " wrench
eCord–
intelligent chip V I. A DDIT IONA L INFO RMAT ION
■■ Solvent quality
Figure 5. eCord intelligent chip identification.
■■ Mobile phase preparation, storage, and age
c. Troubleshooting Questions
b. Getting Started with CORTECS HILIC Columns
1. Are you using 100% aqueous mobile phases?
Note. CORTECS HILIC Columns are designed to retain very polar
bases. Acidic, neutral, and non-polar compounds will have 2. W hat is the age of the mobile phase?
limited retention. 3. Is the mobile phase filtered through a 0.2 µm membrane?
4. Was the mobile phase prepared fresh or topped off?
Mobile Phase Considerations:
1. Always maintain at least 3% polar solvent, such as water, 5. Is the water source of adequate quality?
in the mobile phase, or gradient. T his ensures that the
6. W hen was the last time the water system was serviced or
CORTECS HILIC particle is always hydrated.
was the bottle of water unopened?
2. Maintain at least 40% organic solvent (e.g., acetonitrile)
in your mobile phase or gradient. 7. Is bacterial growth a possibility (pH 7 phosphate buffer is
susceptible to bacterial growth within 24 hours)?
3. Avoid phosphate salt buffers to avoid precipitation in high
organic content mobile phases used for HILIC separations. 8. If a neat standard is prepared in the initial mobile phase
Phosphoric acid is acceptable. conditions and injected, are the problems still observed?
4. Buffered mobile phases such as ammonium formate or 9. If the sample is filtered/purified (i.e., SPE, filtration… etc.) is
ammonium acetate will produce more reproducible results
the problem still observed?
compared to unbuffered additives such as formic acid or acetic
acid. For best peak shape, maintain a buffer concentration 10. Has the quality of the samples changed over time?
of 10 mM.
5. If using an ACQUITY UPLC System, the weak needle wash
solvent should closely match the percent organic solvent V II. C AUT IONA RY NOT E
present in the initial mobile phase conditions, otherwise, Some products may be hazardous during and after use and are
analyte peak shape distortion can occur. to be used by professional laboratory personnel trained in the
competent handling of such materials. The responsibility for the
Injection Solvent Considerations: safe use of products rests entirely with the purchaser and user.
1. Whenever possible, injection solvents should contain 95% The safety data sheets (SDS) for these products are available at
acetonitrile with the polar solvent (i.e., water, methanol, www.waters.com /SDS.
isopropanol) component to be no more than 25% of the total
volume. A generic injection solvent is 75:25 acetonitrile/
methanol. T his is a good compromise between analyte
solubility and peak shape.
Waters Corporation
34 Maple Street
Milford, MA 01757 U.S.A.
Waters, T he Science of W hat’s Possible, CORTECS, UPLC, ACQUITY UPLC, Alliance, Oasis, and Sep-Pak are registered trademarks of Waters
Corporation. eCord and VanGuard are trademarks of Waters Corporation. All other trademarks are the property of their respective owners.
T: 1 508 478 2000
F: 1 508 872 1990
©2016 Waters Corporation. Produced in the U.S.A. September 2016 720005024EN IH-PDF www.waters.com