How do you troubleshoot an HPLC?

Interfering components in sample.

1. Remove guard column (if present) and attempt analysis. Replace guard column if necessary. If analytical column is source of problem, use appropriate restoration procedure (Table 2).
2. Check make-up of mobile phase.
3. Check column performance with standards.

Why does RSD fail in HPLC?

Re: RSD failing HPLC If the RDS of the ratio is substantially worse than that of the individual peaks, that suggests that the major contributions to error are uncorrelated between the peaks, so look at thinks like peak shape, integration settings, baseline noise, etc.

Why sometimes retention problem arises in HPLC?

Increasing back-pressure may indicate a contamination of the column, but even a clogged frit can affect retention times. The pressure needed to push the mobile phase through the frit warms up the mobile phase by friction, and this increase in temperature can affect the retention times.

How can I Fix my HPLC chromatogram problem?

Some of these can be solved by changes in the equipment; however, others require modification of the assay procedure. Selecting the proper column type and mobile phase are keys to “good chromatography.” 1. Blocked frit 1. a. Reverse flush column (if allowed) b. Replace inlet frit 2. Column void 2. Fill void 3. Interfering peak 3. a.

Are there any problems with the HPLC system?

HPLC Troubleshooting: 4. Problems with the Chromatogram 4. Problems with the Chromatogram Many problems in an LC system show up as changes in the chromatogram. Some of these can be solved by changes in the equipment; however, others require modification of the assay procedure.

How to change the pH of a chromatography?

Selecting the proper column type and mobile phase are keys to “good chromatography.” 1. Blocked frit 1. a. Reverse flush column (if allowed) b. Replace inlet frit 2. Column void 2. Fill void 3. Interfering peak 3. a. Use longer column b. Change mobile-phase and/or column/ 4. Wrong mobile-phase pH 4. a. Adjust pH b. For basic compounds, a lower pH