Some Contributions to the Spectroscopy of Hæmoglobin and its Derivatives

Abstract

1. Apparatus.—The following are needed:—Electric or lime light; standard hæmatinometer (i.e., of 1 cm. between the parallel sides) entirely of glass (for many pigments white glass flat-sided stoppered bottles do perfectly well); large direct-vision spectroscope (Messrs Hilger), or a Tollen's carbon-disulphide glass prism (base 5 inches, refracting angle 45°); a well-stretched dead-white screen to receive the spectrum 4 feet long at least, about 12 feet from the lantern.Following Preyer (1), one can with such apparatus first of all demonstrate the different spectroscopic appearance according as blood is or is not diluted. Placing blood newly received from the slaughter-house (defibrinated blood) in the hæmatinometer, we show that it is opaque in a thick layer, no light passes to the prism at all, there is no spectrum, we can say that the percentage of HbO2 is, at least, greater than 7·3. Dilute with pure cold water until a faint gleam of red light appears at the extreme left, the concentration of HbO2 is now not less than 1 per cent.; continue the dilution until the first glimmer of extra-linear green appears, the percentage is now about ·9 per cent.; dilute until the first glimmer of intra-linear green appears, when the percentage is now ·7; add to this its own volume of water, making the percentage fall to ·35. From this point further dilution caused progressive faintness of the two bands, until the vanishing point of the fainter β band is just reached when the hæmoglobin is ·01 per cent.; continue the attenuation until the α band is alone visible, at which point the percentage can be stated to be certainly less than ·01 per cent.