Liquid chromatography tandem mass spectrometry (LC/MS/MS) is widely used in health effects studies due to characteristic sensitivity, accuracy, specificity, dynamic range, ability to identify and quantify large numbers and varieties of analytes, and compatibility with an assortment of matrices. In vivo or in vitro health effects studies often generate small volumes of biofluids that must be split for multiple experiments, which can lead to challenges in achieving method sensitivity. Presented here are LC/MS/MS results of a comparative study of stress response using volumetric absorptive microsampling (VAMS) of whole blood and typically available serum volumes. Health effects related to chemical exposure can be exacerbated with psychosocial and physical stressors, especially for vulnerable populations or life-stages. Prolonged high stress hormone levels occuring during fetal brain development can alter neurodevelopmental function in offspring. Pregnant Long-Evans rats were exposed perinatally to Mn, an environmental neurotoxicant, through drinking water. Half of the rats were also exposed to conditions modeling stressful non-chemical factors. As an indicator of stress response in dams, corticosterone (CORT) was measured at three time points using positive ion electrospray LC/MS/MS. Blood (80 µL) was collected from the lateral tail vein into a serum separator tube. To evaluate potential use of smaller sample sizes, which may minimize stress to animals during sampling, or otherwise provide residual sample volume for additional analyses, a microsample (10 µL) of whole blood was collected at the same time. Detection method sensitivity was enhanced by use of mobile phase additive ammonium fluoride (NH4F). Method optimization and validation parameters will be described. CORT concentrations in whole blood were approximately half those detected from 40 µL serum, and well within detection limits, suggesting VAMS may be an acceptable sampling option for stress response studies.

Measuring stress by LC/MS/MS: Comparison of results from whole blood and serum

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.