What are the primary parameters and acceptance criteria for LC-MS method validation?

What are the primary parameters and acceptance criteria for LC-MS method validation?

LC-MS analysis or Liquid chromatography-mass spectrophotometer is a technique on which industry experts from pioneering biotech companies have relied upon for over 15 years. It is known to drive industry-leading value and turnaround.

It is one of the most common techniques used for conducting DMPK studies. The LC-MS method is a combination of physical separation and detection sensitivity capabilities of both LC and MS platforms. The tandem combination of the two techniques separates analytes into distinct components from a complex matrix along with quantitation of these components.

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The demonstration of the reliability of analytical methods like LC-MS assay is called validation. LC-MS analysis is a highly sophisticated and powerful technique involving various operation modes (MRM, scan, etc), several ionization possibilities (protonation, adduct formation, etc), and numerous parameters.

With numerous possibilities comes a lack of robustness. Hence, the reliability of LC-MS method development must be assured by validating methods.

Some of the parameters and acceptance criteria for LC-MS method validation are selectivity, specificity, sensitivity, linearity, precision, trueness, reproducibility, repeatability, and accuracy.

1. Stability: Estimate method conditions and test the analyte and stability. You should evaluate at least three replicates at two concentration levels under stress conditions.

2. Selectivity: The extent to which one substance interferes with the detection of the other analyte/substance/drug is selectivity. Both specificity and sensitivity of the LC-MS testing are parameters that are used for judging the selectivity.

1. Detector Sensitivity: The lowest level of analytes that can be identified is sensitivity. As MS adds a new dimension to the analyte signal more specific information can be gathered from LC-MS assay.

2. Specificity is defined by verifies that the analyte of interest is detected correctly without cross-reactivity with other related molecules.

3. The separation Factor is one of the best parameters that define how well chromatographic peaks are separated/resolved.

3. Linearity: can be defined in two ways- linear relation between analyte concentration and analyte signal in calibration samples or samples containing matrix components. The factors that can affect the same are ion source, ion transport, and transmission efficiency of the LC-MS analysis.

4. Sensitivity: Determined as the change in the response of an LC-MS assay induced by a change in the gradient calibration or stimulus. It is directly related to the detection/determination ability of the testing. As a threefold parameter sensitivity considers: 1.Daily optimization of instrument parameters

2. Monitoring instrument performance

3. Optimizing for maximum sensitivity of method parameters during method development.

5. Precision: It can be evaluated with a sample that may not have a known analyte and is categorized through the closeness of agreement between measured values, in specified conditions, obtained by replicate measurements on similar objects. Together with accuracy, trueness, and measurement uncertainty, LC-MS method validation can be completed.

FDA accepted parameters or criteria for LC-MS testing generate reliable and robust data for biomarker, tox, or PK studies. As the LC-MS method development combines advantages of HPLC with MS it is often used for a wider variety of drug analysis, food analysis, and environmental testing.