Method of analyzing diluted biological sample component

Abstract

There is provided a method of analyzing a biological sample component that allows easy and accurate quantification and counting of any of a plasma component and a blood cell component in a trace and unknown amount of a whole blood sample collected from a finger, for example. The method of the present invention is a method of analyzing a biological sample component in a trace amount of blood, comprising analyzing a diluent buffer into which the blood has been mixed and an internal standard substance and/or an external standard substance contained in the diluent buffer, calculating a dilution ratio, and analyzing a biological component in a plasma or serum component in the blood.

1 - 31 . (canceled) 
     
     
         32 . A method of quantitative analysis of a component to be analyzed in a blood sample, the method comprising:
 (a) diluting a trace amount of the blood sample with a diluent buffer;   (b) measuring a concentration of an external standard substance in the blood sample diluted with the diluent buffer by absorbance, a Biuret method, a Bradford method, a Lowry method, a bromocresol green method, enzyme methods, using a flame photometer, using atomic absorption, or using an ion selective electrode,   wherein the external standard substance is selected from the group consisting of chloride, albumin and total protein,   wherein the external standard substance is a component homeostatically present comprised at a predetermined concentration in the blood sample prior to the dilution of the blood sample, and   wherein the diluent buffer has a pH 6.5 to 8.0 and is substantially free of the external standard substance; and   (c) calculating the dilution factor of the blood sample from the concentration of the external standard substance measured in (b).   
     
     
         33 . The method of quantitative analysis according to  claim 32 , wherein
 the external standard substance is the chloride.   
     
     
         34 . The method of quantitative analysis according to  claim 32 , wherein
 the diluent buffer comprises:
 an amino alcohol compound selected from the group consisting of 2-amino-2-methyl-1-propanol, 2-ethylaminoethanol, N-methyl-D-glucamine, diethanolamine, and triethanolamine; and 
 a buffering agent selected from the group consisting of HEPES (2-[4-(2-hydoxyethyl)-1-piperazinyl]ethanesulfonic acid), TES (N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid), MOPS (3-morpholinopropanesulfonic acid), and BES (N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid), and 
   wherein the diluent buffer has buffering action at pH 7.4.   
     
     
         35 . The method of quantitative analysis according to  claim 32 , wherein
 the diluent buffer is substantially free of chloride.   
     
     
         36 . A diluent buffer used for diluting a trace amount of the blood sample in the method of quantitative analysis according to  claim 32 , wherein
 the diluent buffer comprises, as buffering agent components, an amino alcohol compound selected from the group consisting of 2-amino-2-methyl-1-propanol, 2-ethylaminoethanol, N-methyl-D-glucamine, diethanolamine, and triethanolamine, and a buffering agent selected from the group consisting of HEPES (2-[4-(2-hydoxyethyl)-1-piperazinyl]ethanesulfonic acid), TES (N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid), MOPS (3-morpholinopropanesulfonic acid), and BES (N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid), and   the diluent buffer has buffering action at pH 7.4.   
     
     
         37 . The buffer according to  claim 32 , which is substantially free of chloride. 
     
     
         38 . The buffer according to  claim 32 , wherein
 the diluent buffer has buffering action at pH 7.4.