Hplc System
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An analyst in HPLC chromatography injects a 5ul and 10 ul injection volume in a valve injector ?
An analyst in hplc chromatography injects a 5ul and 10 ul injection volume into a valve injector and loads the loop manually with a syringe. to save time he decides to use a 10ul loop for both injections. For the 5ul injection he partially fills the loop using a syringe and for the 10ul injection he overloads the loop by loading 15ul into the syringe. He believes that the 15ul will ensure the loop is completely filled and his precision will be better than the 5ul injection. Why did the precision checks show the contrary i.e. the 5ul was better.
Please help me explain why the 5ul was better than 15ul injection into a 10ul loop. Suggest how the analyst could approach this better so the most precise injection of 5 and 10ul samples into the HPLC system. Thanks!
From my personal experience with HPLC, we always get better results by overfilling the loop rather than measuring such small volumes using microliter syringes. Your example suggests the opposite. The main difference I see between your example and what we do in practice is that we usually use 3 or 4 times the loop volume, so we would inject 40 ul in a 10 ul loop. So, displacing the mobile phase in the 10 ul loop with 14 ul of sample will really only leave 5 ul in the loop, and this 5 ul will be less accurate than that measured with the syringe. If one uses 40 ul, and assays 10 ul (loop size), this would be more accurate yet.
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