Filling the Calibrant Bottle in Agilent 6100 Series Single Quadrupole LC/MSD Systems

Answer

This Information Applies To: Agilent 6100 Series LC/MS Instruments

Issue

Instructions on how to fill the calibrant bottle in Agilent 6100 Series LC/MS Instruments when calibrant levels are running low.

Background

The ability to autotune your Agilent LC/MS system relies on providing the instrument with the correct composition and concentration of calibration solution.

This article provides detailed instructions on how to make up your required calibration solution and refill your calibrant bottle in 6100 series LC/MS instruments. 

Items Required

The type of Calibrant used for autotune depends on the type of Ion Source installed on your instrument 

• Clean Glass CDS bottle (p/n 9300-2576), one is already in the instrument
• And one of the calibrants in Table 1 according to the instrument model and installed source type.

Table 1. Calibration solution matrix for the combination of instrument model (left column) and the source type installed on your instrument (top row). 

Steps to Follow

1. From the Acquistion Software, go to the Manual Tune tab, and make sure that the Calibrant on checkbox is cleared (Figure 1). This way, the calibrant bottle is not pressurised by gas when you are unscrewing the bottle.
   

   
   
   Figure 1. Clearing the Calibrant on checkbox
   
   

2. Open the door or remove the cover on the front of the LC/MS to access the calibrant bottle.
   

   Caution: Some force may be required to start the unscrewing of the calibrant bottle. As the bottle is glass, aviod using excess force during removal that may damge the bottle or the bottle cap fixed to the instrument or cause injury. If the bottle cannot be removed easily then double check the calibrant is turned off in MassHunter and then contact your local Agilent service representative for further advice.

3. Gently unscrew the calibant bottle in a counterclockwise direction to remove the bottle from the LC/MS. 

4. Ensure that the calibrant bottle is clean and uncontaminated before refilling the bottle with fresh tuning solution. The bottle can be cleaned using Ultrapure LC/MS Grade Water and an ionic detergent, then rinsed thoroughly with Ultrapure LC/MS Grade Water. Rinse the bottle a minimum of two times with Ultrapure LC/MS Grade Acetonitrile before being used.

5. Confirm the tuning mix that you are using to refill the calibrant bottle is correct for your Ion Source type, application, and mass range. Using the wrong type of tuning mix may cause contamination or will cause autotune failures.
   
   

   Warning: Wear appropriate Personal Protective Equipment including suitable gloves and safety goggles when handling calibration solutions. See the MSDS supplied with the tuning solution for further details.

6. Refill the calibrant bottle about one-third full with the appropriate tuning mix depending on the ion source installed on the instrument (see the Items Required section). The level of solution should be a minimum of 2 cm above the end of the CDS intake tube. The tuning mix is used neat without any dilution steps.

7. Place the intake tube into the refilled calibrant bottle as you lift the calibrant bottle into position, then tighten the bottle in a clockwise direction, see Figure 2.
   
   

   Caution: The bottle only needs to be snug. Do not overtighten the bottle, or you can cause the bottle cap to crack and leak. A leaky bottle cap can prevent the bottle from pressurizing and can lead to the evaporation of the contents.

   
   
   Figure 2. Installing the calibrant bottle into the LC/MS

8. In the Manual Tune tab, check the Calibrant on check box to repressurize the calibrant bottle. See Figure 3

   
   
   Figure 3. Click the check box shown in red to turn on calibrant
   
   

9. Flush the CDS system for a few minutes to ensure that the new tuning solution has reached the nebulizer before trying to tune the instrument. If the autotune still fails after replacing calibrant and checking the nebulizer adjustment and spray shape, then contact your local Agilent support representative for further assistance. 

Learn how to effectively maintain your Agilent 6100 Series LC/MS instrument: LCMS-6100-1101e - Tuning Agilent Single Quadrupole LC/MSD Systems in OpenLab CDS LCMS-6100-1102e - Tuning Agilent Single Quadrupole LC/MSD iQ System in OpenLab CDS e-learning courses available from Agilent education