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Mass Spectrometry Calibration

Mass spectromety is a powerful analytical technique that is used in the biopharmaceutical, pharmaceutical, and medical device industries. It allows the identification and qualification of molecules based on their "mass-to-charge" ratio. In order to obtain reliable and accurate results, calibration of the mass spectrometry is extremely important. The team at GL Technologies has extensive experience in mass spectrometry calibrations for organizations throughout California. We also work with clients to provide mass spectrometry preventive maintenance programs to make sure the instrument is performing as needed and providing reliable results, while also meeting compliance requirements.

To speak with the experts about your mass spec calibration needs, please fill out our online form or give us a call!


Why Calibration is Important?

Calibration is a critical step in mass spectrometry that involves the adjustment of instrument settings to provide and maintain accurate measurements. Calibration is important for a variety of reasons, here are a few of the most important:


Calibration provides precise determination of the mass-to-charge ratios of ions, allowing for the identification and quantification of unknown compounds in complex samples. Accurate calibration makes sure that the instrument is properly adjusted which minimizes systematic errors and improves the overall accuracy of the analysis. By having a reliable calibration curve using known standards, mass spectrometry instruments can be calibrated to accurately measure the masses of ions, which provides scientists with the confidence in their data and facilitating meaningful comparisons across different experiments or laboratories.


By establishing reference points and accurately aligning the mass-to-charge ion scale, calibration corrects for instrument variations including drift and non-linear response, which improves precision. Accurate calibration guarantees the identification and quantification of compouns with exceptional sensitivity and specificity, allowing researchs to make confident scientific inferences.


In the field of quantification, mass spectrometry is extremely important. Accurate quantification of analysis relies on the ability of a mass spectrometry to measure the masses of ions and determine their abundance. Calibration makes sure that the instrument is properly adjusted to consistently measure mass-to-charge ratios, which results in reliable and reproducible quantification. With mass spectrometry calibration, the instrument's sensitivity, linearity, and accuracy can be improved, allowing for precise determination of analyte concentrations. The calibration process establishes a foundation for quantitative analysis to allow research to confidently measure trace amounts of compounds, monitor changes, and identify subtle differences in complex samples.

Methods of Mass Spectrometry Calibration

There are a few popular methods that are used for mass spectrometry calibration, here are the most common approaches.

External Calibration

External calibration is a widely used method for calibrating mass spectrometry. In this method, a set of known standard compounds with well-characterized mass-to-charge values is introduced into the mass spectrometer. The standards include small molecules, peptides, or proteins, depending on the required calibration range. By analyzing the spectra of the standard compounds, the instrument's mass scale is adjusted to accurately assign mass-to-charge values to unknown samples. This approach offers versatility, allowing for routine calibration checks for the best instrument performance.

Internal Calibration

With internal calibration, a reference compound is introduced into the sample matrix before or during the analysis. This serves as an internal standard for calibration purposes. The reference compound should possess well-defined ionizable species that are distinct from the analytes of interest. By monitoring the signal of the reference compound (which can be isotopically labeled), the instrument response can be accurately determined and any variations can be corrected. Internal calibration provides a convenient means to achieve high precision in mass spectrometry measurements.

Matrix-Matched Calibration

Matrix matched calibration involves preparing calibration standards that closely resemble the matrix of the samples to be analyzed. This technique is advantageous when dealing with complex samples containing diverse matrices. In this method, teh calibration standards are prepared by spiking a blank matrix with known concentrations of the analytes of interest. By using this calibration curve, which considers the specific mtarix effects, accurate quantifiation of the analytes can be achieved. This method accounts for the potential interferences caused by the matrix which improves the precision and accuracy of the mass spec analysis.

Multipoint Calibration

This approach to calibration involves creating a calibration curve by analyzing a series of reference standards with known concentrations. The reference standards are selected to cover a broad range of masses or mass-to-charge ratios, providing comprehensive coverage fo the mass spectrometer's detection capabilities. By plotting the instrument's response, like peak intensity against the known concentrations, a calibration curve is generated. The curve is then used to accurately quantify and identify unknown samples by comparing their response to the calibration curve. This method not only accounts for instrument variability but also allows for corrections of non-lineary responses.

Maintenance and Verification

Calibration is not a 1 time process; regular maintenance and verification are required for maintaining instrument performance.

Routine Calibration

Routine calibration plays an important role in this process as it involves the systematic verification and adjustment of instrument paramaters to uphold measurement precision. During a routine calibration, mass spectometer compones such as ion sources, detectors, and voltage supplies are examined and calibrated using certified reference materials. This process allows for identification and correction of any deviations so the instrument remains within specified tolerances. Laboratories need to adhere to a well-defined routine cailbration schedule to maintain their mass spectrometry instruments.

Quality Control Samples

In this maintenance process, quality control samples play a role by providing a means to monitor and verify the performance of mass spectrometers over time. The samples are carefully prepared with known concentrations of target compounds, serve as benchmarks for instrument calibration and to validate the accuracy of the system. Regular analysis of quality control samples provides the detection and correction of any instrument drift or systematic errors, providing reliability of mass spectrometry measurements.

Performance Checks

With mass spec calibration maintenance, there needs to be regular perfromance checks to make sure analytical results are accurate. These checks work to maintain the instrument's sensitivity over time and the process includes verifying the mass accuracy and resolution, evaluating ion intensity stability and background levels. These performance checks need to be performed at regular intervals so researchs can identify and address any inconsistencies to maintain continued quality of mass spectrometry measurements.

Preventive Maintenance

Preventive maintenance is an important practice that maintains peak performance, accuracy, and reliability of mass spectrometers. This proactive approach involves a series of systematic inspections, calibrations, and maintenance taks to fix any potential issues before they escalate. Regular cleaning and verification of instrument components including ion sources, vacuum systems, and detectors, works to maintain peak performance and sensitivity. Along with this, software updates, data backups, and routine performance checks aid in preventing instrument downtime and data loss. It's recommended that laboratories have a comprehensive preventive maintenance schedule in place to maximize the longevity of their mass spectrometers and minimize potential unexpected failures.

To speak with the experts about your mass spec calibration needs, please fill out our online form or give us a call!


GL Technologies Capabilities

As a full-service company specializing in equipment calibration, repair, and certification services for biopharmaceutical, pharmaceutical, and medical device industries. Our team has extensive experience working with sPRT calibrations along with CMMS softwareHPLC OQ validation, and fume hood certifications. Companies of all sizes rely on our team to implement, maintain, and keep their research and manufacturing processes compliant with regulatory standards. Other specialties include building maintenance systems, and mass spectrometry calibrations.  Our team works with clients throughout San Diego, Los Angeles, and San Francisco covering the entire State of California with our services!


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