Care and use manual – Waters ACQUITY UPLC CSH130 C18 1.7 μm and XP 2.5 μm Columns User Manual

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[ CARE AND USE MANUAL ]

ACQUITY UPLC CSH130 C

Columns

VI. ADDITIONAL INFORMATIONA.

Tips for maximizing ACQUITY UPLC CSH130 C

Column Lifetimes

1. To maximize ACQUITY UPLC CSH130 C

column lifetime, pay close

attention to:

• Water quality (including water purification system)

• Solvent quality

• Mobile-phase preparation, storage and age

• Sample, buffer and mobile-phase solubilities

• Sample quality and preparation.

2. When problems arise, often only one improper practice must be changed.

3. Always remember to:

• Use in-line filter unit or, preferably, a VanGuard Pre-column.

• Discourage bacterial growth by minimizing the use of 100%
aqueous mobile phases where possible.

• Change aqueous mobile phase every 24-48 hours (if 100%
aqueous mobile phase use is required).

• Discard old 100% aqueous mobile phases every 24-48 hours
to discourage bacterial growth.

• Add 5%-10% organic modifier to mobile phase A and adjust
gradient profile.

• Filter aqueous portions of mobile phase through 0.2 µm filter.

• Maintain your water purification system so that it is in good
working order.

• Only use ultra pure water (18 megohm-cm) water and highest quality
solvents possible. HPLC grade water is not UPLC grade water.

• Consider sample preparation (e.g., solid-phase extraction, filtration, etc).

4. Avoid (where possible):

• 100% aqueous mobile phases (if possible).

• HPLC-grade bottled water.

• “Topping off” or adding “new” mobile phase to “old” mobile phase.

• Old aqueous mobile phases. Remember to rinse bottles thoroughly and
prepare fresh every 24-48 hours.

• Using phosphate salt buffer in combination with high ACN concentrations
(e.g., > 70%) due to precipitation.

5. Don’t: assume a “bad” column is the culprit when high back pressure or split
peaks are observed. Investigate cause of column failure:

• Backpressure

• Mobile phase(s), bacteria, precipitation and/or samples

• Peak splitting

• Sample quality

• Injection solvent strength.

6. Remember: UPLC flow rates are often much lower and, therefore,
mobile phases last much longer (only prepare what you need or store
excess refrigerated).

7. Mobile-phase related questions to ask:

• Am I using 100% aqueous mobile phases? Am I able to add
a small amount of organic modifier to my mobile-phase A?

• Do I filter my aqueous mobile phases through 0.2 µm filters?

• How old is my mobile phase? Do I label the bottle with preparation date?

• Do I “top off” or do I prepare fresh mobile phases every 24-48 hours?

• What is the quality of my water? Has the quality recently changed? How
is my water purification system working? When was it last serviced?

• Am I working with a pH 7 phosphate buffer (which is VERY susceptible
to bacterial growth)?

8. Sample-related questions to ask:

• If I inject neat standards prepared in mobile phase do I observe
these problems?

• If I prepare my standards in water and prepare them like samples
(e.g., SPE, filtration, etc.) do I still observe these problems?

• Has the quality of my samples changed over time?