By Jim on Friday, September 29, 2000 - 09:30 am:
What is the best way to determine the gradient dwell time (tD) for an HPLC system? Should one just estimate it from the sums of all the tubing volumes between the mixer and the column? Thanks
By Tom M. on Friday, September 29, 2000 - 02:55 pm:
Jim,
We measure it as part of our calibration. Water in Ch. A, 0.5% Acetone in Ch. B., no column. Program a 0-100% B gradient in 20 min., 1mL/min. wavelength set at 272 nm.
You might need some backpressure to ensure that the check valves are operating properly. You can use a backpressure regulator or a length of cap. tubing.
Draw a tangent from the linear part of the gradient back to the baseline where they intersect is the dwell time. Good luck.
By Jim W on Monday, October 2, 2000 - 06:41 am:
Thanks Tom!
By Anonymous on Sunday, June 27, 2004 - 09:23 pm:
dear Jim/Tom
could u please tell me how to calculate dwell time during LC/MS-MS analysis.how it affects the response of anlyte.
By Smitaa on Sunday, June 27, 2004 - 09:47 pm:
Dear freinds
i would like to have a discussion on dwell time calculations during bioanalytical method development on LC/MS-MS.
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