Waters ACQUITY UPLC HSS Columns - Manual (Page 8)

[ Care and Use ManUal ]
XSelect HSS HPlc columnS 8
b. Recommended Flow Rates and Backpressures for Reversed-Phase ACQUITY UPLC HSS Columns
1.0 mm i.d. Columns (40 °C)
UPLC Linear Velocity
(mm/sec)
3 4 5 6
Column Dimensions
Flow Rate
(mL/min)
Backpressure
(psi)
Flow Rate
(mL/min)
Backpressure
(psi)
Flow Rate
(mL/min)
Backpressure
(psi)
Flow Rate
(mL/min)
Backpressure
(psi)
1.0 x 50 mm 0.1 3800 0.13 5600 0.17 6500 0.2 7600
1.0 x 100 mm 0.1 7500 0.13 9800 0.17 12800 0.2 15100
1.0 x 150 mm 0.1 11300 0.13 14600 0.17 19100 0.2 25500
2.1 mm i.d. Columns (40 °C)
UPLC Linear Velocity
(mm/sec)
3 4 5 6
Column Dimensions
Flow Rate
(mL/min)
Backpressure
(psi)
Flow Rate
(mL/min)
Backpressure
(psi)
Flow Rate
(mL/min)
Backpressure
(psi)
Flow Rate
(mL/min)
Backpressure
(psi)
2.1 x 30 mm 0.45 2700 0.60 3600 0.75 4500 0.9 5400
2.1 x 50 mm 0.45 4200 0.60 5700 0.75 7100 0.9 5800
2.1 x 100 mm 0.45 8000 0.60 10700 0.75 13400 0.9 16100
2.1 x 150 mm 0.45 11800 0.60 15800 0.75 19700 0.9 23700
Note: 1) ACQUITY HSS UPLC 1.8 µm particle reversed-phase columns
2) ACN/Aqueous gradient, Pmax at ~30% ACN
3) Approximate maximum total system backpressure given.
3.0 mm i.d. Columns (40 °C)
UPLC Linear Velocity
(mm/sec)
3 4 5 6
Column Dimensions
Flow Rate
(mL/min)
Backpressure
(psi)
Flow Rate
(mL/min)
Backpressure
(psi)
Flow Rate
(mL/min)
Backpressure
(psi)
Flow Rate
(mL/min)
Backpressure
(psi)
3.0 x 30 mm 0.9 3100 1.17 4000 1.53 5300 1.8 6200
3.0 x 50 mm 0.9 4600 1.17 6000 1.53 7800 1.8 9200
3.0 x 100 mm 0.9 8300 1.17 10800 1.53 14100 1.8 16700
3.0 x 150 mm 0.9 12000 1.17 15700 1.53 20500 1.8 24100
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 hrs?
• 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 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?