Apparatus for measuring lipid peroxidation in biological fluids and suspensions of tissues

Abstract

The present invention relates to a method and apparatus for measuring the lipid peroxidation extent in biological fluids and suspensions of tissues, in which a specifically prepared lipids containing sample is exposed to heat so as to induce thermochemiluminescence light which is emitted by the sample and is enhanced to an amount which can be detected by a special purpose photo detector. The measurement of the light allows to conclude about the content of peroxides and subsequently to obtain accurate assessment of lipid peroxidation.

We claim:  
     
         1 . A method of measuring lipid peroxidation in biological fluids, in suspensions of biological tissues or the like, wherein a sample of said fluid or suspension is heated so as to induce therein thermochemical luminescence (TCL) and amount of said TCL can be measured, said method comprising the following sequence of steps: 
 a) bringing a sample of said fluid or suspension in a receptacle having substantially flat bottom    b) placing said receptacle within a sublimation chamber and putting thereof on a substrate made of substantially metallic material    c) imparting reciprocating motion to said receptacle so as to distribute said sample over the bottom of said receptacle in a substantially homogeneous manner    d) subjecting of said sample residing within said chamber to sublimation at reduced pressure during period of time sufficient for evacuation from said sample of a liquid phase and formation on the bottom of said receptacle of a substantially continuous film consisting of dry solid residual    e) bringing said receptacle with said residual in a processing chamber and placement thereof on a plate having temperature sufficient for inducing TCL and emitting thereof by said residual    f) sensing said TCL and measuring the intensity thereof for example by optical photodetector during period of time sufficient for establishing stable correlation between TCL intensity and time.    
     
     
         2 . Method as defined in  claim 1 , in which said receptacle is formed as disposable vessel made of thin metallic foil and said plate is made of aluminum.  
     
     
         3 . Method as defined in  claim 1 , in which said reciprocating motion is imparted to said sample during at least 40 sec so as to distribute said sample over the bottom of said receptacle as a thin, substantially continuous layer.  
     
     
         4 . Method as defined in  claim 1 , in which said sublimating step is carried out at reduced pressure not exceeding 2 mbar and during period of time not exceeding 10 min.  
     
     
         5 . Method as defined in  claim 1 , in which before bringing said receptacle into said processing chamber said plate is heated up to at least 60 deg. C.  
     
     
         6 . Method as defined in  claim 1 , in which said TCL intensity is measured in the range of 400-600 nm during at least 300 sec.  
     
     
         7 . An apparatus for measuring lipid peroxidation in biological fluids, in suspensions of biological tissues or the like, wherein a sample of said fluid or suspension is heated so as to induce therein thermo-chemical luminescence (TCL) and amount of said TCL can be measured, said apparatus comprising: 
 a) sublimation chamber provided with a means for homogeneous distributing of said sample within a receptacle and with a sublimating means capable to evacuate liquid phase from said sample so as to form a thin substantially continuous film consisting of dry solid residual, said means for homogeneous distributing comprises a substrate made of preferably metallic material, said substrate is provided with a depression for receiving said receptacle therein and there is provided a driving means for imparting reciprocating motion to said substrate    b) heating chamber provided with a plate for placing said receptacle thereon, with a heating means for heating said plate and with a means for detection and measuring said TCL    c) data recording unit for presentation in visual manner results of measuring of said TCL    d) control and data processing unit for controlling said means for homogeneous distributing, said driving means, said sublimating means, said heating means, said means for detection and measuring said TCL and said data recording unit.    
     
     
         8 . Apparatus as defined in  claim 7 , in which said sublimating means comprises vacuum pump capable to generate reduced pressure within said sublimation chamber.  
     
     
         9 . Apparatus as defined in  claim 7 , in which said substrate is formed as a disc made of aluminum, said disc is supported by elastic tubular elements connecting said disc with the basis of said sublimating chamber and said driving means comprises a motor imparting reciprocating motion to said plate via a cam.  
     
     
         10 . Apparatus as defined in  claim 7 , in which said heating means comprises transistor and temperature sensor and said means for detection and measuring TCL comprises photo-electronic multiplier.