“…[22][23][24] Herein, we take advantage of the unique optical shift of Hb's Soret band and combine it with partial chemical deoxygenation to optically measure Hb-O 2 dissociation under physiologically acidic conditions at pH 6.9. First, we harnessed the acidic pH environment to facilitate rapid chemically induced partial deoxygenation of Hb (increased Bohr effect), 1,25 to reduce Hb deoxygenation time down to 90 seconds. We have shown that, during this process, the conformational changes in Hb's tertiary structure result in significant shifts in the optical absorption spectra which concede with Circular Dichroism (CD) changes.…”