Review of the Use of Liquid Chromatography-Tandem Mass Spectrometry in Clinical Laboratories: Part I-Development

Abstract

The process of method development for a diagnostic assay based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) involves several disparate technologies and specialties. Additionally, method development details are typically not disclosed in journal publications. Method developers may need to search widely for pertinent information on their assay(s). This review summarizes the current practices and procedures in method development. Additionally, it probes aspects of method development that are generally not discussed, such as how exactly to calibrate an assay or where to place quality controls, using examples from the literature. This review intends to provide a comprehensive resource and induce critical thinking around the experiments for and execution of developing a clinically meaningful LC-MS/MS assay.

Validation of a Microwave-Assisted Derivatization Gas Chromatography-Mass Spectrometry Method for the Quantification of 2-Hydroxybutyrate in Human Serum as an Early Marker of Diabetes Mellitus

“…However, there are few validated methods for the determination of this metabolite in serum samples. Although mass spectrometry may be considered the gold standard method for the measurement of metabolites, quantitative methods based on mass spectrometry also require careful validation to provide objective evidence that the method is valid for the intended use and that it exhibits appropriate performance characteristics [ 20 , 21 ]. In our case, we performed a double validation of the method following the EMEA guideline [ 22 ], with two different internal standards, 2HB-d 3 and 3HB-d 4 .…”

Review of the Use of Liquid Chromatography-Tandem Mass Spectrometry in Clinical Laboratories: Part II–Operations

“…High-purity water is an excellent diluent for LC-MS/MS assays. When solvents are used for dilution, adequate equilibration times for the mixtures and ISs established in validation must be included in the SOP [ 3 ].…”

“…This review is a follow-up of a previous one [ 3 ]. Together, these publications intend to capture the lifecycle of LC-MS/MS from development through validation to the execution and maintenance of a clinical assay.…”

Back to the Basics of Liquid Chromatography-Mass Spectrometry

“…In practice, reproducing results of mass spectrometric analyses by referring to one or two method development research articles is challenging, mainly because researchers lack background knowledge, and articles often do not include detailed and necessary background knowledge. In this issue, the review article by Rappold [ 5 ], written from the author’s experience and other literature, will be of great help to researchers working on LC-MS/MS method development. This review article enlists the necessary elements for LC-MS/MS method development, which will aid researchers develop their own methods.…”

“…To that end, Rappold has provided a valuable and complete account of all aspects of LC-MS/MS method development, including reagent selection, sample preparation, LC and MS conditions, calibration, quality control, and optimization. This review article by Rappold [ 5 ] is replete with helpful and practical information for researchers, technicians, and laboratory physicians working with LC-MS/MS.…”