Mass spectrometry has been a crucial component of laboratory and industrial analysis since the middle twentieth century. For several decades, the requirement of sophisticated techniques and specialized understanding had confined the applications of mass spectrometers to specialized laboratories. However, simplifying technology and operational protocols have made this instrument more accessible to a broad range of users.
Although mass spectrometry applications are dominated by complex protein characterization and biomarker discovery studies, they are employed increasingly in routine clinical assessments. Hence, mass spectrometers are rapidly combined with other technologies, such as High-Performance Liquid chromatography (HPLC), to expand their applications further. The current article discusses the role of mass spectrometry in modern HPLC labs.
The significance of mass spectrometry in HPLC testing services
Bioanalytical laboratories play a crucial role in accelerating biomedical and clinical research. They have expertise in systems and technologies such as PK/PD testing, immunogenicity assessments, and neutralizing antibody assays. Similarly, HPLC labs are critical in offering HPLC testing services for numerous biomedical and clinical applications.
Today, HPLC systems are coupled with mass spectrometry to separate and study analytes in complex study matrices. The liquid chromatography component separates the compounds based on the interaction with the stationary and mobile phases, while the MS unit detects them based on their mass-to-charge ratio. This method improves assay specificity by segregating interferences such as isobaric compounds. Importantly, the strength of LC-MS systems is due to their inherently high analytic selectivity. Hence, mass spectrometers play a significant role in HPLC laboratories. Let us explore some of the roles of HPLC systems in detail.
LC-MS testing plays a critical role in screening neonatal genetic diseases. Neonatal dried blood spots can be tested to assess the possibility of several inborn metabolism errors. The advent of electrospray ionization techniques further accelerated the application of LC-MS in processing large sample volumes of neonatal blood spots.
The high cost of commercial assays for therapeutic drug monitoring has led to a growth in the development of LC-MS-based alternatives. Today, LC-MS assays are developed for immunosuppressants such as sirolimus, cyclosporine, everolimus, mycophenolic acid, and tacrolimus. Besides, LC-MS systems for anticancer drugs, antiretrovirals, and aminoglycosides are also available. The primary advantage of LC-MS systems in therapeutic drug monitoring is their ability to multiplex so that several drugs can be studied in a single assay run, simplifying workflows and providing additional data.
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Commercial immunoassays give variable responses to different Vitamin D forms and their metabolites. Hence, LC-MS systems have garnered considerable interest in vitamin D measurements. Several research studies have used LC-MS assays to evaluate 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 in serum using electrospray ionization. Besides, the derivatization technique has improved the sensitivity of metabolites at low levels, such as salivary, urine, and 1,25- and 24,25-dihydroxy metabolites. Additionally, LC-MS assays for other fat-soluble vitamins, for example, different forms of vitamin 15, E13, and vitamin K15, and water-soluble vitamins such as folate vitamers, riboflavin, and pyridoxine are developed. One such assay has multiplexing capacities to study different forms of vitamins A, D, E, and K.
In Conclusion
mass spectrometry combined with HPLC systems has massive potential to accelerate biomedical and clinical research.
