The invention discloses a novel method for measuring
hydrogen pumping capability of mitochondria, and belongs to the field of scientific research and experiments. The method includes S1) putting a
magnetic stirrer below an
absorption cell rack of a spectrophotometer; S2) setting the spectrophotometer into a time scanning state; S3) adding an
absorption cell into the spectrophotometer, adding KCl into the
absorption cell, starting the
magnetic stirrer and starting scanning; S4) adding 8-hydroxy-1,3,6-pyrenetrisulfonic acid; S5) adding ferrocytochrome c, valinomycin, 2,3-dimethoxy-5-methyl-6-(10-bromo)-1,4-
benzoquinone and a
mitochondrion solution; S6) recording an absorption value A1 after an absorption line is flat; S7) adding 1 [mu]L of
sodium ferricyanide having a concentration of 5 mM,and recording an absorption value A2; S8) adding a
proton carrier that is [(3-chlorophenyl)hydrazono]
malononitrile, and recording an absorption value A3; and S9) adding
sodium ferricyanide, and recording an absorption value A4; and S10) calculating the
hydrogen pumping capability according to an equation that is (H<+> / e)=x / y=(A1-A2) / (A3-A4). The method is reasonable in design, simple to operate and low in interference by the environment, and a measurement result is good in precision and
repeatability. The method is suitable for use in common laboratories.