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Development of a Method and Validation for the Quantitation of FruArg in Mice Plasma and Brain Tissue Using UPLC–MS/MS
journal contribution
posted on 2016-10-31, 00:00 authored by Mitch
C. Johnson, Hailong Song, Jiankun Cui, Valeri V. Mossine, Zezong Gu, C. Michael GreenliefAged garlic extract (AGE) is a popular
nutritional supplement and
is believed to promote health benefits by exhibiting antioxidant and
anti-inflammatory activities and hypolipidemic and antiplatelet effects.
We have previously identified N-α-(1-deoxy-d-fructos-1-yl)-l-arginine (FruArg) as a major contributor
to the bioactivity of AGE in BV-2 microglial cells whereby it exerted
a significant ability to attenuate lipopolysaccharide-induced neuroinflammatory
responses and to regulate the Nrf2-mediated antioxidant response.
Here, we report on a sensitive ultraperformance liquid chromatography–tandem
mass spectrometry (UPLC–MS/MS) protocol that was validated
for the quantitation of FruArg in mouse plasma and brain tissue samples.
Solid-phase extraction was used to separate FruArg from proteins and
phospholipids present in the biological fluids. Results indicated
that
FruArg was readily absorbed into the blood circulation of mice after
intraperitoneal injections. FruArg was reliably detected in the subregions
of the brain tissue postinjection, indicating that it penetrates the
blood–brain barrier in subnanomolar concentrations that are
sufficient for its biological activity.
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attenuate lipopolysaccharide-induced neuroinflammatory responsesNrf 2-mediated antioxidant responsebrain tissue samplesintraperitoneal injectionsUPLCBV -2 microglial cellsFruArgSolid-phase extractionmouse plasmahealth benefitsMice Plasmaantiplatelet effectsbrain tissue postinjectionBrain Tissueblood circulationanti-inflammatory activitiessubnanomolar concentrations
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