SOP for Cleaning of High Performance Liquid Chromatography (HPLC)

1.0 OBJECTIVE

To clearly lay down the procedure for cleaning.

2.0 SCOPE

This procedure is applicable for of all high performance liquid chromatography installed in quality control laboratory.

3.0 RESPONSIBILITY

3.1 Doing: Technical Assistant
3.2 Checking: Executive/Manager

4.0 ACCOUNTABILITY

Head of the Department

5.0 PROCEDURE

5.1 For system cleaning
5.1.1 After completion of analysis, clean the HPLC as follows.
5.1.2 Transfer the used glassware for washing & keep aside test tube stands lying near the HPLC system. Clean the surrounding area with tissue paper.
5.1.3 Clean the syringe with repeated rinses of distilled water.
5.1.4 Clean the outer surface of injector by flushing the distilled water with syringe forcefully near the injector hole and joints of loops. Clean it with tissue paper.
5.1.5 Clean the inner surface of injector and loops by injecting distilled water into loop with syringe three to four times. ( Note: for normal phase system use non polar solvents like Acetonitrile, Methylene chloride, n-Hexane, Tetrahydrofuran for cleaning)
5.1.6 Clean the plunger seal by injecting distilled water three to four times with syringe.
5.1.7 Remove the column and replace with joints for the cleaning of tubings & flush with 6N nitric acid, distilled water and then with methanol.
5.1.8 Remove the air in suction pipe by purging the line and flushing with distilled water for some time.5.2 Clean the suction filter by sonicating it into nitric acid for 10 to 15 minutes and then wash it with water.
5.3 Replace the joints with column and wash the column first with distilled water and then with methanol.5.4 Switch “OFF” the pump, detector and integrator.
5.5 Clean the outer surface of instrument with dry clean cloth.
5.6 Remove any dirt or spot with isopropyl alcohol.
5.7 Dry it with tissue paper.

5.8 Frequency:
1. General cleaning: Daily
2. Syringe, loop and injector plunger seal: after completion of each analysis
3. Flow cell, pipeline, suction filter: when problem arise
5.9 Keep the record of cleaning.

6.0 ABBREVIATIONS

6.1 N=Normal
6.2 HPLC = High performance liquid chromatography
6.3 S.O.P.= Standard operating procedure

Related: Principle of High Performance Liquid Chromatography

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