The Importance of Mass Spectrometry
Mass spectrometry is a powerful analytical tool used in the biopharmaceutical industry to determine the identity and quantity of protein and peptide therapeutics. However, accurate quantification of these complex molecules can be challenging due to variability in the mass spectrometer's performance over time. To ensure reliable results, regular calibration of the mass spectrometer is essential.
Calibration is the process of verifying and adjusting the accuracy of the mass spectrometer. Calibration ensures that the instrument is performing as expected and producing accurate and reproducible results. Calibration is critical for biopharmaceuticals because even small errors in mass measurement can have significant consequences in determining the identity and purity of the drug product.
First Step in Mass Spec Calibration
The first step in mass spectrometry calibration is to select an appropriate calibration standard. Calibration standards are compounds of known mass that are used to calibrate the instrument. Common calibration standards used in biopharmaceutical mass spectrometry include peptides, proteins, and small molecules. It is important to select a calibration standard that is similar in mass and chemical composition to the analyte of interest.
Once the calibration standard is selected, the mass spectrometer is tuned to optimize its performance. The mass spectrometer is adjusted to ensure that it is properly detecting and measuring ions of the calibration standard. This is done by adjusting the ionization source, ion optics, and detector settings. The tuning process is essential for obtaining accurate and precise mass measurements.
Second Step in Mass Spec Calibration
Next, the calibration standard is introduced into the mass spectrometer, and a mass spectrum is acquired. The mass spectrum is a plot of ion signal intensity versus mass-to-charge ratio (m/z). The m/z values of the ions in the mass spectrum are compared to the known masses of the calibration standard. The mass spectrometer is then calibrated by adjusting the instrument's settings to match the known masses of the calibration standard.
Calibration must be performed regularly to ensure that the mass spectrometer is operating within the acceptable range. The frequency of calibration depends on the instrument's stability and the nature of the samples being analyzed. In general, it is recommended to perform calibration before each sample analysis to ensure the most accurate results.
In conclusion, mass spectrometry calibration is a critical step in the analysis of biopharmaceuticals. Calibration ensures that the mass spectrometer is producing accurate and reliable results, which is essential for determining the identity and purity of protein and peptide therapeutics. By selecting an appropriate calibration standard, optimizing the instrument's performance, and regularly calibrating the mass spectrometer, biopharmaceutical researchers can obtain the most accurate and reliable results possible.