I have a Shimadzu LC 20AT HPLC with Prominence SPD 20A UV-VIS detector. For a few days I have been facing a problem in the UV detector. Whatever the mobile phase I use or whatever be the concentration of the drug injected there is no change in the baseline. A straight line only comes continuously. I tried to change the column, mobile phase, increased the drug concentration but to no avail. Kindly suggest something to overcome this problem.
Hi Bhupendra,
This might be silly but just to be sure, check whether the out-let tube from the column is in deed going to the detector. If it's connected, you will have to consider using other detectors just to confirm that the column and everything else is ok. I will also try a different sample or a standard analyte to see that the detector is working.
Gud luck...
Hi,
I would also try another sample (e.g. a standard). Maybe the lamp of your detector is at the end of its lifetime. Another possibility would be, your drug sample sticks to the column. What mobile phases did you use?
Suggest to check the optical fiber connection (if it is possible in SPD 20A UV-VIS), since your description seems exactly like behavior of mine FPLC with this part removed.
The fractions you obtained check with the any spectrophotometer suitable, to be sure it is the problem. Also the flow-cell can be dirty.
There is some thechnical problem. If you know troubleshooting then try above suggestions otherwise call the engineer.
I have tried different samples, different mobile phases such as methanol/water 50:50, ACN:water 50:50, methanol with phosphate buffer but there is no change in baseline. There is actually some baseline noise but it is very small and continuous which does not change at all.
Yes the flow is all right and there is no leaks in the tubing and the purge valve is also closed. The pressure of the system is also stable and correct. Now I will inject some organic solvents and check otherwise I should infer that the detector lamp has to be replaced. Thanks everyone for your valuable comments.
- Badges
- Science method