Method and an apparatus for determining blood components

• US 4,997,769 A
• Filed: 04/13/1990
• Issued: 03/05/1991
• Est. Priority Date: 06/21/1985
• Status: Expired due to Term

• Current Assignee: Radiometer A/S (Danaher Corporation)
• Original Assignee: Radiometer A/S (Danaher Corporation)
• Inventors: Lundsgaard, Finn C.
• Primary Examiner(s): WARDEN, JILL ALICE
• Assistant Examiner(s): Johnston, Jill
• Application Number:
  US07/512,038
• Time in Patent Office: 326 Days
• Field of Search: 436/47, 436/67, 436/66, 422/67, 422/82.09, 422/73, 422/81, 356/39, 356/40, 356/41
• US Class Current: 436/66
• CPC Class Codes: G01N 2021/3129 Determining multicomponents by multiwavelength light
• CPC Class Codes: G01N 21/31 Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry G01N21/72 takes precedence
• CPC Class Codes: Y10T 436/113332 with conveyance of sample along a test line in a container or rack

Abstract

Method and apparatus for spectrophotometrically determining the concentration of a number of hemoglobin derivatives comprising at least deoxyhemoglobin and oxyhemoglobin or parameters derived from the concentration of individual hemoglobin derivatives, in particular the oxygen saturation fraction, in whole blood, in which light is transmitted to the blood under test, the modification on the light transmitted to the blood which the blood causes is determined at a number of individual wavelengths, and the concentrations (or the derived parameters) are determined on the basis of the light modification at the individual wavelengths and on the basis of predetermined coefficients representing the light absorption characteristics of each of the hemoglobin derivatives at each of the individual wavelengths, any error on the determination values due to turbidity being substantially compensated for by utilizing predetermined coefficients representing an absorption characteristic of turbidity at each of the wavelengths. The measurement is normally performed at a number of wavelengths which is at least equal to the number of hemoglobin derivatives to be determined plus one. Although the turbidity-caused contribution to the total reduction in the light transmission is not based on any direct proportionality between the reduction in light transmission and the concentration of the turbidity-causing components (such as is the case with the hemoglobin derivatives), the "absorbances" caused by the turbidity at each particular wavelength may simply be eliminated from the equation set from which the concentrations of the hemoglobin derivatives are calculated.

Claims

1. A method for spectrophotometrically determining the concentration of a number of hemoglobin derivatives comprising at least deoxyhemoglobin and oxyhemoglobin or parameters derived from the concentration of individual hemoglobin derivatives, including the oxygen saturation fraction, in whole blood, comprising

transmitting light to the blood under test,

determining the modification caused by the blood on the light transmitted to the blood at a number of individual wavelengths, wherein one of the individual wavelengths is above 535 nm and is a wavelength at which the ratio between absorption due to turbidity and absorption due to hemoglobin derivative is relatively high, and

determining the concentrations or the derived parameters on the basis of the light modification at the individual wavelengths and on the basis of predetermined coefficients representing the light absorption characteristics of each of the hemoglobin derivatives at each of the individual wavelengths, any error on the determination values due to turbidity being substantially compensated for by utilizing predetermined coefficients representing an absorption characteristic of turbidity at each of the individual wavelengths.

2. The method of claim 1, wherein the wavelength is above 650 nm.

3. The method of claim 1, wherein the measurement is performed at a number of individual wavelengths which is at least equal to the number of hemoglobin derivatives to be determined plus one.

4. The method of claim 1, including determining the concentrations of any further components which modify the absorbance caused by the light transmitted to the blood sample at one or more of said individual wavelengths, or of the parameters derived from the concentrations of any of said further components, on the basis of said light absorbance modifications and on the basis of sets of predetermined coefficients for said further components applied to the measured absorbances of the blood sample at each said individual wavelength in an equation for the determination of the concentrations of said further components derived from the Lambert-Beer'"'"'s Law;

wherein each set of predetermined coefficients for said further components has been calculated using the Lambert-Beer'"'"'s Law from the concentration and actual measured light absorbance of a known solution of each of said further components at each said individual wavelength, said individual hemoglobin derivatives, turbidity components and further components being treated as equivalent factors in the blood sample in deriving the foregoing equations for the determination of the concentrations of said further components, and in calculation of said coefficients for said further components.

5. The method of claim 4, wherein said further components comprise at least one diagnostic dyestuff selected from the group consisting of Evan'"'"'s blue, Methylene Blue and Cardi Green, and/or at least one plasma pigment selected from the group consisting of bilirubin and carotene.

6. The method of claim 1, wherein the hemoglobin derivatives whose concentration is determined comprise deoxyhemoglobin, oxyhemoglobin, and at least one hemoglobin derivative selected from the group consisting of carboxyhemoglobin, methemoglobin and sulfhemoglobin.

7. The method of claim 1, wherein the light transmitted to the blood sample includes at least said individual wavelengths;

and wherein the light, after absorbance modification by the blood sample is divided into components at least including said individual wavelengths.

8. A method for spectrophotometrically determining the concentration of a number of hemoglobin derivatives comprising at least deoxyhemoglobin and oxyhemoglobin or parameters derived from the concentration of individual hemoglobin derivatives, including the oxygen saturation fraction, in whole blood, comprising

transmitting light to the blood under test,

determining the modification caused by the blood on the light transmitted to the blood at a number of individual wavelengths, wherein the measurement is performed at a number of wavelengths which is at least equal to the number of hemoglobin derivatives to be determined plus one, and

determining the concentrations or the derived parameters on the basis of the light modification at the individual wavelengths and on the basis of predetermined coefficients representing the light absorption characteristics of each of the hemoglobin derivatives at each of the individual wavelengths, any error on the determination values due to turbidity being substantially compensated for by utilizing predetermined coefficients representing an absorption characteristic of turbidity at each of the individual wavelengths.

9. The method of claim 8, wherein one of said individual wavelengths at which the absorbance modification caused by the blood on the light transmitted to the blood sample is determined at a wavelength above 650 nm at which the ratio between the absorbance due to the turbidity component and the absorbance due to said individual hemoglobin derivatives is relatively high.

10. The method of claim 9, including determining the concentrations of any further components which modify the absorbance caused by the light transmitted to the blood sample at one or more of said individual wavelengths, or of the parameters derived from the concentrations of any of said further components, on the basis of said light absorbance modifications and on the basis of sets of predetermined coefficients for said further components applied to the measured absorbances of the blood sample at each said individual wavelength in an equation for the determination of the concentrations of said further components derived from the Lambert-Beer'"'"'s Law;

wherein each set of predetermined coefficients for said further components has been calculated using the Lambert-Beer'"'"'s Law from the concentration and actual measured light absorbance of a known solution of each of said further components at each said individual wavelength, said individual hemoglobin derivates, turbidity components and further components being treated as equivalent factors in the blood sample in deriving the foregoing equations for the determination of the concentrations of said further components, and in calculating said coefficients for said further components.

11. The method of claim 10, wherein said further components comprise at least one diagnostic dyestuff selected from the group consisting of Evan'"'"'s blue, Methylene Blue and Cardio Green, and/or at least one plasma pigment selected from the group consisting of bilirubin and carotene.

12. The method of claim 8, wherein the hemoglobin derivatives whose concentration is determined comprise deoxyhemoglobin, oxyhemoglobin, and at least one hemoglobin derivative selected from the group consisting of carboxyhemoglobin, methemoglobin and sulfhemoglobin.

13. The method of claim 8, wherein the light transmitted to the blood sample includes at least said individual wavelengths;

and wherein the light, after absorbance modification by the blood sample is divided into components at least including said individual wavelengths.

14. A method for spectrophotometrically determining the concentration of deoxyhemoglobin, oxyhemoglobin and one or several hemoglobin derivatives selected from carboxyhemoglobin, methemoglobin and sulfhemoglobin, in whole blood, comprising

transmitting light to the blood under test,

determining the modification caused by the blood on the light transmitted to the blood at a number of individual wavelengths, wherein the measurement is performed at a number of individual wavelengths, and

determining the concentrations or the derived parameters on the basis of the light modification at the individual wavelengths and on the basis of predetermined coefficients representing the light absorption characteristics of each of the hemoglobin derivatives at each of the individual wavelengths, any error on the determination values due to turbidity being substantially compensated for by utilizing predetermined coefficients representing an absorption characteristic of turbidity at each of the individual wavelengths.

15. The method of claim 14, wherein one of said individual wavelengths at which the absorbance modification caused by the blood on the light transmitted to the blood sample is determined at a wavelength above 650 nm at which the ratio between the absorbance due to the turbidity component and the absorbance due to said individual hemoglobin derivatives is relatively high.

16. The method of claim 14, including determining the concentrations of any further components which modify the absorbance caused by the light transmitted to the blood sample at one or more of said individual wavelengths, or of the parameters derived from the concentrations of any of said further components, on the basis of said light absorbance modifications and on the basis of sets of predetermined coefficients for said further components applied to the measured absorbances of the blood sample at each said individual wavelength in an equation for the determination of the concentrations of said further components derived from the Lambert-Beer'"'"'s Law;

wherein each set of predetermined coefficients for said further components has been calculated using the Lambert-Beer'"'"'s Law from the concentration and actual measured light absorbance of a known solution of each of said further components at each said individual wavelength, said individual hemoglobin derivatives, turbidity components and further components being treated as equivalent factors in the blood sample in deriving the foregoing equations for the determination of the concentrations of said further components, and in calculation of said coefficients for said further components.

17. The method of claim 16, wherein said further components comprise at least one diagnostic dyestuff selected from the group consisting of Evan'"'"'s blue, Methylene Blue and Cardio Green, and/or at least one plasma pigment selected from the group consisting of bilirubin and carotene.

18. The method of claim 14, wherein the light transmitted to the blood sample includes at least said individual wavelengths;

and wherein the light, after absorbance modification by the blood sample is divided into components at least including said individual wavelengths.

19. A method of spectrophotometrically determining the concentration of a number of individual hemoglobin derivatives comprising at least deoxyhemoglobin and oxyhemoglobin or parameters derived from the concentration of individual hemoglobin derivatives, including the oxygen saturation fraction, in whole blood, with substantial compensation for any error in the values of the individual concentrations or parameters caused by at least one turbidity component in the whole blood, comprising;

(a) transmitting light to a blood sample which is under test;

(b) determining the absorbance modification caused by the blood on the light transmitted to the blood sample at a number of individual wavelengths which are at least equal to the number of hemoglobin derivatives to be determined plus the number of turbidity components being compensated for;

(c) determining the values of said individual hemoglobin derivative concentrations or of said parameters derived from said individual hemoglobin derivative concentrations on the basis of said light absorbance modifications and on the basis of sets of predetermined coefficients applied to the measured absorbances of the blood sample at each wavelength in an equation for the determination of the concentration of each of said derivatives derived from the Lambert-Beer'"'"'s Law, wherein each set of predetermined coefficients has been calculated using the Lambert-Beer'"'"'s Law from the concentration and actual measured light absorbance of a known solution of each individual hemoglobin derivative and turbidity component at each of said individual wavelengths, said individual hemoglobin derivatives and turbidity components being treated as equivalent factors in the blood sample in deriving the foregoing equations for the determination of the concentrations of said derivatives, and in calculating said coefficients.

20. The method of claim 19, wherein one of said individual wavelengths at which the absorbance modification caused by the blood on the light transmitted to the blood sample is determined at a wavelength above 650 nm at which the ratio between the absorbance due to the turbidity component and the absorbance due to said individual hemoglobin derivatives is relatively high.

21. The method of claim 19, including determining the concentrations of any further components which modify the absorbance caused by the light transmitted to the blood sample at one or more of said individual wavelengths, or of the parameters derived from the concentrations of any of said further components, on the basis of said light absorbance modifications and on the basis of sets of predetermined coefficients for said further components applied to the measured absorbances of the blood sample at each said individual wavelength in an equation for the determination of the concentrations of said further components derived from the Lambert-Beer'"'"'s Law;

wherein each set of predetermined coefficients for said further components has been calculated using the Lambert-Beer'"'"'s Law from the concentration and actual measured light absorbance of a known solution of each of said further components at each said individual wavelength, said individual hemoglobin derivatives, turbidity components and further components being treated as equivalent factors in the blood sample in deriving the foregoing equations for the determination of the concentrations of said further components, and in calculation of said coefficients for said further components.

22. The method of claim 21, wherein said further components comprise at least one diagnostic dyestuff selected from the group consisting of Evan'"'"'s blue, Methylene Blue and Cardio Green, and/or at least one plasma pigment selected from the group consisting of bilirubin and carotene.

23. The method of claim 19, wherein the hemoglobin derivatives whose concentration is determined comprise deoxyhemoglobin, oxyhemoglobin, and at least one hemoglobin derivative selected from the group consisting of carboxyhemoglobin, methemoglobin and sulfhemoglobin.

24. The method of claim 19, wherein the light transmitted to the blood sample includes at least said individual wavelengths;

and wherein the light, after absorbance modification by the blood sample is divided into components at least including said individual wavelengths..

25. An apparatus for spectrophotometrically determining the concentration of a number of hemoglobin derivatives in a blood sample comprising at least deoxyhemoglobin and oxyhemoglobin or parameters derived from the concentration of individual hemoglobin derivatives in whole blood including the oxygen saturation fraction, said apparatus comprising

light source means for transmitting to the blood under test,

sample inlet means,

a conduit for conducting the blood sample from the sample inlet means through the apparatus, the conduit having transparent wall parts defining a light transmission path through a sample positioned in the conduit,

hemolyzing means for hemolyzing the blood sample in the conduit, said hemolyzing means including ultrasonic sound generation means, the light source means emitting light to the light transmission path,

light detector means for receiving light transmitted through the blood sample, and

measuring means for determining the modification of the transmitted light at a number of individual wavelengths and for determining the concentrations or the derived parameters on the basis of the modification of transmitted light measured at the individual wavelengths and on the basis of predetermined coefficients representing characteristics of the light absorption of each of the hemoglobin derivatives at each of the individual wavelengths,

said measuring means comprising computer means further including:

(i) storage means for storing data representing the predetermined coefficients of a number of hemoglobin derivatives at each of the individual wavelengths and for storing data representing the coefficients representing the absorption characteristic of turbidity at each of the individual wavelengths and

(ii) calculating means for calculating the oxygen saturation fraction, the hemoglobin derivative concentration or the derived parameters on the basis of the absorption of the blood sample measured at the individual wavelengths and the said data, ny error on the determinations due to turbidity being substantially compensated for by utilizing predetermined coefficients representing an absorption characteristic of turbidity at each of the individual wavelengths.

26. The apparatus of claim 25, wherein the light source means to adapted generate light including a plurality of wavelengths, and whrein the apparatus further comprises

means for dividing the light transmitted through the blood sample into individual components, at least including said individual wavelengths.

27. An apparatus for spectrophotometrically determining the concentration of a number of individual hemoglobin derivatives in a blood sample comprising at least deoxyhemoglobin and oxyhemoglobin or parameters derived from the concentration of individual hemoglobin derivatives, including the oxygen saturation fraction, in whole blood, with substantial compensation for any error in the values of the individual concentrations or parameters caused by at least one turbidity component in the whole blood, said apparatus comprising

(a) light source means for transmitting light to a blood sample which is under test;

(b) light detector means for receiving light modified by the blood sample; and

(c) measuring means for determining

(i) the absorbance modification caused by the blood on the light transmitted to the blood sample at a number of individual wavelengths which are at least equal to the number of hemoglobin derivatives to be determined plus the number of turbidity components being compensated for; and

(ii) the values of said individual hemoglobin derivative concentrations or of said parameters derived from said individual hemoglobin derivative concentrations on the basis of said light absorbance modifications and on the basis of sets of predetermined coefficients applied to the measured absorbances of the blood sample at each wavelength in an equation for the determination of the concentration of each of said derivatives derived from the Lambert-Beer'"'"'s Law, wherein each set of predetermined coefficients has been calculated using the Lambert-Beer'"'"'s Law from the concentration and actual measured light absorbance of a known solution of each individual hemoglobin derivative and turbidity component at each of said individual wavelengths, said individual hemoglobin derivatives and turbidity components being treated as equivalent factors in the blood sample in deriving the foregoing equations for the determination of the concentrations of said derivatives, and in calculating said coefficients.

28. The apparatus of claim 27, further comprising:

(d) sample inlet means

(e) a conduit for conducting the blood sample from the sample inlet means through the apparatus, the conduit having transparent wall parts defining a light transmission path through a sample positioned in the conduit,

the light source means emitting light to the light transmission path and the light detector means for receiving light transmitted through the blood sample.

29. The apparatus of claim 28, wherein the measuring means comprises computer means further including:

(i) storage means for storing data representing the predetermined coefficients of a number of hemoglobin derivatives at each of the individual wavelengths and for storing data representing the coefficients representing the absorption characteristic of turbidity at each of the individual wavelengths, and

(ii) calculating means for calculating the oxygen saturation fraction, the hemoglobin derivative concentrations or the derived parameters on the basis of the absorbance of the blood sample measured at the individual wavelengths and the said data.

30. The apparatus of claim 27, wherein the light source means is adapted to generate light including a plurality of wavelengths, and wherein the apparatus further comprises

(f) means for dividing the light transmitted through the blood sample into individual components, at least including said individual wavelengths.

31. The apparatus of claim 27, further comprising

(g) hemolyzing means for the blood sample present within the conduit, including ultrasonic sound generator means.