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Characterisation and comparison of temporal release profiles of nitric oxide generating donors.

Bradley SA, Steinert JR - J. Neurosci. Methods (2015)

Bottom Line: We found that donors such as NOC-5 and PAPA-NONOate decayed substantially within days, whereas SNP and GSNO showed greater stability releasing consistent levels of NO over days.In all donors tested, the amount of released NO differs between frozen and unfrozen stocks.Fluorescent and amperometric approaches to measure NO concentrations yield a wide range of levels.

View Article: PubMed Central - PubMed

Affiliation: MRC Toxicology Unit, Hodgkin Building, University of Leicester, Leicester LE1 9HN, UK.

No MeSH data available.


Related in: MedlinePlus

Temporal release of NO from NOC-5. (A) Average release profile of NO yield from fresh NOC-5 stock at 5, 10 and 20 μM over 30 min. (B) Average NO release profiles on day two at 5, 10 and 20 μM over a 30 min recording period. (C) Average profiles of NO release over 30 min from three day old stock at 5, 10 and 20 μM. (D) Average NO release profiles of 4 week old frozen NOC-5 stock at 5, 10 and 20 μM over a 30 min recording period. (E) Box and whisker plots showing the plateau concentration range of NO recorded using NOC-5 stock on day one, two and three and frozen stock. (F) Box and whisker plots depicting the area under the release curve using the same stock over 3 consecutive days and frozen stock.
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fig0020: Temporal release of NO from NOC-5. (A) Average release profile of NO yield from fresh NOC-5 stock at 5, 10 and 20 μM over 30 min. (B) Average NO release profiles on day two at 5, 10 and 20 μM over a 30 min recording period. (C) Average profiles of NO release over 30 min from three day old stock at 5, 10 and 20 μM. (D) Average NO release profiles of 4 week old frozen NOC-5 stock at 5, 10 and 20 μM over a 30 min recording period. (E) Box and whisker plots showing the plateau concentration range of NO recorded using NOC-5 stock on day one, two and three and frozen stock. (F) Box and whisker plots depicting the area under the release curve using the same stock over 3 consecutive days and frozen stock.

Mentions: NOC-5 is a diazeniumdiolate compound that belongs to the same family of NO donors as PAPA NONOate. As such, this donor possesses all of the favourable qualities of PAPA NONOate with the addition of a slightly longer half-life of 93 min at 22 °C, pH 7.4 compared to 77 min for PAPA NONOate. Accordingly, NO release from NOC-5 stock over three consecutive days resulted in a greater NO release over 30 min. In a similar manner to PAPA NONOate, all concentrations of fresh NOC-5 in PBS led to a rapid increase in NO release on day one. A steady NO release plateau was reached at ∼17 min at 5 μM, ∼14 min at 10 μM and ∼12 min at 20 μM (Fig. 4A). On days two and three NO release was slower resulting in shallower curves and longer plateau times of ∼25 and ∼26 min (Fig. 4B and C). Frozen stock liberated less NO compared to fresh stocks (Fig. 4D). NO release was greatest on day one with 5 μM yielding a plateau concentration of 330 ± 37 nM, 10 μM: 480 ± 49 nM and 20 μM: 657 ± 84 nM NO. By day two the amount of NO released at plateau has decreased by half (5 μM: 131 ± 24 nM, 10 μM: 217 ± 38 nM and 20 μM: 352 ± 68 nM) and furthermore by day three (5 μM: 86 ± 30 nM, 10 μM: 141 ± 48 nM and 20 μM: 230 ± 76 nM). Frozen stock gave plateau concentrations of NO that range between that seen at day two and three in fresh stock (5 μM: 103 ± 9 nM, 10 μM: 179 ± 8 nM, 20 μM: 290 ± 20 nM) (Fig. 4D, Table 1). Total NO release over the 30 min (AUC) recording period decreased with concentration and time (Fig. 4F).


Characterisation and comparison of temporal release profiles of nitric oxide generating donors.

Bradley SA, Steinert JR - J. Neurosci. Methods (2015)

Temporal release of NO from NOC-5. (A) Average release profile of NO yield from fresh NOC-5 stock at 5, 10 and 20 μM over 30 min. (B) Average NO release profiles on day two at 5, 10 and 20 μM over a 30 min recording period. (C) Average profiles of NO release over 30 min from three day old stock at 5, 10 and 20 μM. (D) Average NO release profiles of 4 week old frozen NOC-5 stock at 5, 10 and 20 μM over a 30 min recording period. (E) Box and whisker plots showing the plateau concentration range of NO recorded using NOC-5 stock on day one, two and three and frozen stock. (F) Box and whisker plots depicting the area under the release curve using the same stock over 3 consecutive days and frozen stock.
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fig0020: Temporal release of NO from NOC-5. (A) Average release profile of NO yield from fresh NOC-5 stock at 5, 10 and 20 μM over 30 min. (B) Average NO release profiles on day two at 5, 10 and 20 μM over a 30 min recording period. (C) Average profiles of NO release over 30 min from three day old stock at 5, 10 and 20 μM. (D) Average NO release profiles of 4 week old frozen NOC-5 stock at 5, 10 and 20 μM over a 30 min recording period. (E) Box and whisker plots showing the plateau concentration range of NO recorded using NOC-5 stock on day one, two and three and frozen stock. (F) Box and whisker plots depicting the area under the release curve using the same stock over 3 consecutive days and frozen stock.
Mentions: NOC-5 is a diazeniumdiolate compound that belongs to the same family of NO donors as PAPA NONOate. As such, this donor possesses all of the favourable qualities of PAPA NONOate with the addition of a slightly longer half-life of 93 min at 22 °C, pH 7.4 compared to 77 min for PAPA NONOate. Accordingly, NO release from NOC-5 stock over three consecutive days resulted in a greater NO release over 30 min. In a similar manner to PAPA NONOate, all concentrations of fresh NOC-5 in PBS led to a rapid increase in NO release on day one. A steady NO release plateau was reached at ∼17 min at 5 μM, ∼14 min at 10 μM and ∼12 min at 20 μM (Fig. 4A). On days two and three NO release was slower resulting in shallower curves and longer plateau times of ∼25 and ∼26 min (Fig. 4B and C). Frozen stock liberated less NO compared to fresh stocks (Fig. 4D). NO release was greatest on day one with 5 μM yielding a plateau concentration of 330 ± 37 nM, 10 μM: 480 ± 49 nM and 20 μM: 657 ± 84 nM NO. By day two the amount of NO released at plateau has decreased by half (5 μM: 131 ± 24 nM, 10 μM: 217 ± 38 nM and 20 μM: 352 ± 68 nM) and furthermore by day three (5 μM: 86 ± 30 nM, 10 μM: 141 ± 48 nM and 20 μM: 230 ± 76 nM). Frozen stock gave plateau concentrations of NO that range between that seen at day two and three in fresh stock (5 μM: 103 ± 9 nM, 10 μM: 179 ± 8 nM, 20 μM: 290 ± 20 nM) (Fig. 4D, Table 1). Total NO release over the 30 min (AUC) recording period decreased with concentration and time (Fig. 4F).

Bottom Line: We found that donors such as NOC-5 and PAPA-NONOate decayed substantially within days, whereas SNP and GSNO showed greater stability releasing consistent levels of NO over days.In all donors tested, the amount of released NO differs between frozen and unfrozen stocks.Fluorescent and amperometric approaches to measure NO concentrations yield a wide range of levels.

View Article: PubMed Central - PubMed

Affiliation: MRC Toxicology Unit, Hodgkin Building, University of Leicester, Leicester LE1 9HN, UK.

No MeSH data available.


Related in: MedlinePlus