How to Calibrate your HPLC - Agilent 1220 Manual Injector
Calibrating the HPLC
Purpose
This document details how to properly calibrate an Agilent 1220 Manual Injection HPLC to deliver accurate results for cannabis potency testing.
Responsibility
It is the responsibility of the client to ensure that the calibrant kit being used is not expired and has been properly stored in a freezer at or below -20 °C.
Materials and Equipment Materials:
Calibrant Kit (Includes 100 uL of sample - estimated 4 to 6 injections per vial) 1 ug/mL Standard (A Labeled Amber Vial)
5 ug/mL Standard (B Labeled Amber Vial)
10 ug/mL Standard (C Labeled Amber Vial)
5 ug/mL Standard (B Labeled Amber Vial)
10 ug/mL Standard (C Labeled Amber Vial)
25 ug/mL Standard (D Labeled Amber Vial) 50 ug/mL Standard (E Labeled Amber Vial)
Equipment:
Agilent 1220 Manual Injection HPLC system 50 uL syringe
Safety Issues
Safety glasses and disposable nitrile gloves should always be worn when handling chemicals. In the event of skin contact, eye contact, ingestion, or inhalation, refer to the appropriate MSDS for first aid measures.
Procedure
1. Remove the calibration standards from the -20 °C freezer and let them equilibrate at room temperature for 3 minutes.
2. Make sure the HPLC is on and warmed up so that it is ready to run samples. a. Run through the Startup Method if the HPLC is not ready to run samples.
3. At the Cann-ID home page, click the blue "Calibrate" button.
4. Click “Start Test” on screen.
5. Using the syringe needle, wash the needle port 8 times with 20 uL of wash solution.
6. After washing the injection port, wipe the outside of the syringe tip with a Kimwipe that is wet with a small amount of wash solution. Remove any wash solution that may still be present in the syringe by repeatedly drawing up and pushing down the syringe plunger with the tip positioned over a dry Kimwipe.
8. Use the 50 uL needle to puncture the seal of the calibrant vial.
9. Put the needle inside the standard glass vial, and draw up at least 15 uL.
4. Click “Start Test” on screen.
5. Using the syringe needle, wash the needle port 8 times with 20 uL of wash solution.
6. After washing the injection port, wipe the outside of the syringe tip with a Kimwipe that is wet with a small amount of wash solution. Remove any wash solution that may still be present in the syringe by repeatedly drawing up and pushing down the syringe plunger with the tip positioned over a dry Kimwipe.
8. Use the 50 uL needle to puncture the seal of the calibrant vial.
9. Put the needle inside the standard glass vial, and draw up at least 15 uL.
Important: Make sure the lowest bubble is above the 15 uL line. 10. Follow the prompt on Cann-ID and inject the sample as directed. Be sure that the air bubble does NOT get injected.
11. Throw the lever only once and keep the lever down throughout the duration of the run.
12. Leave the syringe in place until the lever is in the down position and then remove the
syringe.
13. After the analysis has finished and the program is prompting you to inject the next calibrant vial, return the lever to the upright position and rinse the injection port again with 8 rinses of ~50 microliters of blue sticker wash.
14. Again, after washing the injection port, wipe the outside of the syringe tip with a Kimwipe that is wet with a small amount of wash solution. Remove any wash solution that may still be present in the syringe by repeatedly drawing up and pushing down the syringe plunger with the tip positioned in a dry Kimwipe.
14. Again, after washing the injection port, wipe the outside of the syringe tip with a Kimwipe that is wet with a small amount of wash solution. Remove any wash solution that may still be present in the syringe by repeatedly drawing up and pushing down the syringe plunger with the tip positioned in a dry Kimwipe.
15. Inject the next standard.
16. Repeat steps 5-15 for each of the standards (A-E Vials).
17. Upon completion, customers must contact the Ionization Labs science team to develop their
16. Repeat steps 5-15 for each of the standards (A-E Vials).
17. Upon completion, customers must contact the Ionization Labs science team to develop their
calibration table and update their method.For instructions on how to check the calibration status of your HPLC, please reference the attached.
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