Table 1.
Chromatographic conditions for analysis of free CoA (CoASH) and acetylCoA (unlabelled, +4 and +6 AMU) in brain extracts.
Table 2.
MRM conditions for analysis of free CoA (CoASH) and acetylCoA (unlabelled, +4 and +6 AMU) in brain extracts.
Fig 1.
Levels of free CoA (CoASH), acetylCoA and pantothenic acid (PA) in male mouse A) liver and B) brain after administration of a low PA diet for 3 and 6 days.
*p < 0.02; N.S. No Significance. Error bar, mean ± SD for triplicate samples.
Fig 2.
Level of unlabelled pantothenic acid (PA), free CoA (CoASH) and acetylCoA in A) female and B) male mouse liver and C) female and D) male brain after administration of a single oral dose of [13C3-15N]-PA. C57BL/6N mice were kept for 3 days under a PA-free diet and then orally dosed with 25 mg/kg of [13C3-15N]-PA. Liver and brain samples were harvested over a collection period of 0–342 h and unlabelled PA, CoASH and acetylCoA level were measured by LC-MS.
Table 3.
Average endogenous level of pantothenic acid (PA), free CoA (CoASH) and acetylCoA in the male and female C57BL/6N mouse liver and brain.
Fig 3.
Half-life of free CoA (CoASH) and acetylCoA in A) female B) male mouse liver. C57BL/6N mice were kept for 3 days under a PA-free diet and then orally dosed with 25 mg/kg of [13C3-15N]-PA. Liver samples were collected and analysed by LC-HRMS to determine labelled (+4 AMU) and unlabelled CoASH and acetylCoA levels. The metabolite half-life was calculated from Cmax to the last time point collected. Error bars, mean ± SD for triplicate samples.
Fig 4.
Half-life of free CoA (CoASH) and acetylCoA in A) female B) male mouse brain. C57BL/6N mice were kept for 3 days under a PA-free diet and then orally dosed with 25 mg/kg of [13C3-15N]-PA. Brain samples were collected and analysed by LC-HRMS to determine the labelled (+4 AMU) and unlabelled CoASH and acetylCoA levels. The metabolite half-life was calculated from Cmax to the last time point collected. Error bars, mean ± SD for triplicate samples.
Table 4.
Summary of [13C3-15N]-pantothenic acid ([13C3-15N]-PA), free CoA (CoASH) (+4 AMU) and acetylCoA (+4 AMU) half-life in male and female C57BL/6N mouse brain and liver.
Fig 5.
Formation of labelled-CoA species from labelled-fosmetpantotenate (+6 AMU).
Table 5.
Average endogenous level of unlabelled free CoA (CoASH), acetylCoA and total CoA in male C57/Bl6 mouse brain (n = 27) during the intrastriatal infusion study over a period of 336 h after dosing.
Fig 6.
Half-life in male mouse brain of A) free CoA (CoASH) (+4 and+6 AMU), B) acetylCoA (+4 and +6 AMU) and C) total CoA +4 and +6 AMU. C57/Bl6 mice received an intrastriatal injection of 125 μg of +6 AMU labelled-fosmetpantotenate at each brain hemisphere (250 μg total dose) and brain samples were collected over a period of 336 h after dosing. For CoASH +4, acetylCoA +4 and total CoA +4 the time interval 96–336 h was considered for the half-life calculation. For CoASH +6, acetylCoA +6 and total CoA +6 the time interval 24–336 h was used and t = 48 h was excluded from the regression. Error bars, mean ± SD for triplicate samples.
Table 6.
Summary of half-life in mouse brain for free CoA (CoASH), acetylCoA and total CoA (+4 and +6 AMU) after striatal injection of +6 AMU labelled-fosmetpantotenate.