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LC-MS Calibration vs. Tuning

This Edition of HPLC solutions is comparing the purpose and procedure between LC-MS calibration and tuning..
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John Dolan
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In this technical article you will learn about the two procedures that are used to make sure that an HPLC equipped with a mass spectral detector (MS) is reporting proper mass information and that the system is properly set to detect the analyte(s) of interest.

These are calibration and tuning – two processes that are sometimes confused. Calibration is the process used to ensure mass accuracy; tuning is used to make sure that the instrument is working well for a particular sample. Most commonly for quantitative analysis, a single- (LC-MS) or triple- (LC-MS/MS) quadrupole MS detector is used, although the principles discussed here also hold for other LC-MS detectors. Calibration gives us confidence that when the detector reports a mass (or more properly mass-to-charge ratio, m/z ) for a compound, we can be sure that the reported value is correct. This means that we need to use a well-defined standard for the procedure. Tuning is a much more common process than calibration. The essence of tuning is that we want the instrument to give the maximum signal possible for our analyte. To do this, we use the same instrumental setup as for calibration but with a solution of our analyte or internal standard instead of the calibrant.

By reading the full article you will find out more about these two processes and how calibration ensures that the instrument is reporting accurate masses, whereas tuning is used to make sure that adequate signal intensity is obtained for specific analytes.

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Meet the Author(s):

  • John Dolan

    John Dolan is considered to be one of the world’s top experts in HPLC. He has written more than 300 user-oriented articles on HPLC troubleshooting over the last 30 years, in addition to more than 100 peer-reviewed technical articles on HPLC and related techniques. His three books (co-authored with Lloyd Snyder), Troubleshooting HPLC Systems, Introduction to Modern Liquid Chromatography (3rd edn), and High-Performance Gradient Elution, are standard references on thousands of desks around the world. He has taught HPLC training classes around the world to more than 10,000 students.

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