Rotenone: Difference between revisions
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== Application in [[HRR]] == | == Application in [[HRR]] == | ||
ย | {{Chemical_description | ||
:::'''Rot: Rotenone''' (C<sub>23</sub>H<sub>22</sub>O<sub>6</sub>); Sigma R 8875, 1 g, store commercial stock at RT; M = 394.42 gยทmol<sup>-1</sup> (โฅ 95%). | |abbr=Rot | ||
ย | |trivial name=Rotenone | ||
:::: <span style="color:#8B008B"> '''Caution:''' Very toxic and light sensitive (store solution in a dark glass vial).</span> | |complete name= | ||
|chem formula=C<sub>23</sub>H<sub>22</sub>O<sub>6</sub> | |||
|molar mass=394.42 | |||
|vendor=Sigma-Aldrich | |||
|product number=R8875 | |||
|store at=RT | |||
|sensitivity=light sensitive | |||
|cas=83-79-4 | |||
|h statements=H300, H330, H315, H319, H335, H410 | |||
|h info=fatal if swallowed, fatal if inhaled, causes skin irritation, causes serious eye irritation, may cause respiratory irritation, very toxic to aquatic life with long-lasting effects}}<!--:::'''Rot: Rotenone''' (C<sub>23</sub>H<sub>22</sub>O<sub>6</sub>); Sigma R 8875, 1 g, store commercial stock at RT; M = 394.42 gยทmol<sup>-1</sup> (โฅ 95%). | |||
:::: <span style="color:#8B008B"> '''Caution:''' Very toxic and light sensitive (store solution in a dark glass vial).</span>--> | |||
:::: <span style="color:#8B008B"> '''Caution:''' In solution and exposed to light and air, rotenone decomposes losing toxicity within days, see [https://pubchem.ncbi.nlm.nih.gov/compound/6758#section=Stability-Shelf-Life PubChem] for details.</span> | |||
Revision as of 10:48, 10 June 2021
Description
Rotenone is an inhibitor of Complex I (CI) and thus inhibits NADH oxidation. It inhibits the transfer of electrons from iron-sulfur clusters in CI to ubiquinone via binding to the ubiquinone binding site of CI. See also Complex II-linked substrate state, S(Rot).
Abbreviation: Rot
Reference: Gnaiger 2020 BEC MitoPathways, MiPNet09.12, MiPNet03.02
Application in HRR
- Rot: Rotenone ( C23H22O6), Sigma-Aldrich: R8875, store at RT (light sensitive), CAS: 83-79-4, M = 394.42 gยทmol-1
- Hazard statements: H300, H330, H315, H319, H335, H410; fatal if swallowed, fatal if inhaled, causes skin irritation, causes serious eye irritation, may cause respiratory irritation, very toxic to aquatic life with long-lasting effects
- Rot: Rotenone ( C23H22O6), Sigma-Aldrich: R8875, store at RT (light sensitive), CAS: 83-79-4, M = 394.42 gยทmol-1
- Caution: In solution and exposed to light and air, rotenone decomposes losing toxicity within days, see PubChem for details.
- Preparation of 1 mM stock solution (dissolved in 99.9 % ethanol) for 2-mL O2k-chamber:
- Weigh about 1 mg of rotenone.
- Calculate the amount of ethanol abs. needed: 1 mL of ethanol abs. per 0.39 mg of rotenone.
- Rotenone does not dissolve easily in ethanol. Warming to 35ยฐC and an ultrasonic bath will improve solubility.
- Divide into 100 ยตL aliquots (use dark glass vials).
- Store frozen at -20 ยฐC.
- ยป O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (2-mL O2k-chamber): 1 ยตL using a 10 ยตL Hamilton syringe.
- Final concentration: 0.5 ยตM.
- Preparation of 0.25 mM stock solution (dissolved in 99.9 % ethanol) for 0.5-mL O2k-chamber:
- Take 50 ยตL of the 1 mM stock solution.
- Add 150 ยตL of ethanol absolute (99.9 %).
- ยป O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (0.5-mL O2k-chamber): 1 ยตL using a 10 ยตL Hamilton syringe.
- Final concentration: 0.5 ยตM.
- Microalgae applications
- For the specific case of microalgal cells it is recommended to prepare higher concentration:
- Preparation of 5.5 mM stock solution (dissolved in 99.9 % ethanol)
- Weigh 4.4 mg of rotenone and dissolve in 2 ml of ethanol absolute (99.9 %).
- Divide into 200 ยตl aliquots (use dark glass vials).
- Store frozen at -20 ยฐC.
- ยป O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (2-mL O2k-chamber): 15 ยตL using a 25 ยตL Hamilton syringe.
- Experimental concentration: 41 ยตM.
Inhibition of CI by rotenone
- Fato R, Bergamini C, Bortolus M, Maniero AL, Leoni S, Ohnishi T, Lenaz G (2009) Differential effects of mitochondrial Complex I inhibitors on production of reactive oxygen species. Biochim Biophys Acta 1787:384โ92. >>PMC2724837
History
- Up to 2006, rotenone was added at a high final concentration (0.5 ยตM), hence a 0.5 mM stock solution was prepared. Since 0.05 ยตM may be fully inhibiting, we suggested to use a stock solution at lower concentration (0.1 mM), to reduce the problem of rotenone retention. The 2006 edition listed a 0.2 mM stock solution, but the amount added to 10 ml ethanol was given for a 0.15 mM stock solution. Our recent rotenone titrations with permeabilized muscle fibers, however, confirmed that the originally proposed higher rotenone concentration is actually required for full inhibition.
MitoPedia topics:
Inhibitor