5. Normal and pathological color characteristics of human blood components

The quantitative colorimetric characteristics of hemolyzed blood, plasma, and

The experiment. The objects of investigation were solutions of hemolyzed blood and solutions of plasma and serum (prepared from that blood) of the same concentration from 100 male and female donors (in different blood groups and age groups) and from 95 patients who were assigned to three arbitrary groups: (I) 41 patients with purulent diseases (osteomyelitis, purulent fistulas, gonitis), (II) 41 resuscitated patients (acute myocardial infarction, acute cerebral circulatory collapse, chronic cardiac insufficiency), and (III) 13 patients with cirrhosis of the liver. We determined the color characteristics of the "average" donor (without separating the donors according to blood, sex, and age groups) and compared them with the analogous characteristics of patients from the different groups. The blood for the studies was drawn at a blood donation center and in clinical departments by stan-

The spectra of the solutions of hemolyzed blood, plasma, and serum of concentration 2.5 vol.% (1.40) were taken in quartz cuvets with thickness of the working layer of liquid equal to 10 mm at room temperature, on an SF-2000 spectrophotometer in the range 200–1000 nm in 20 nm steps. As the solvent and the reference solution, we used distilled water for injection, which is optically neutral under the experimental conditions and is the natural physiological solvent in the human body. The hemolyzed blood was prepared using a standard heparin solution, and the

In addition to the averaged values of the color coordinates and the lightness value, we calculated the standard deviation, the confidence interval for significance

Figure 8 shows the averaged spectra for the hemolyzed blood, plasma, and

corresponding averaged spectra of the donors. There are clear differences between

For the plasma and serum, over the entire studied region, the group spectra for the patients lie higher than the averaged spectra of the donors, and their positional order is consistent: the averaged spectrum for patients with purulent diseases lies above the averaged spectrum for the donors, the averaged spectrum for the

Spectra of hemolyzed donor blood (1), plasma (2), and serum (3) in the UV and visible regions (averaged over

serum from the patients in all three examined groups compared with the

plasma was prepared using the preservative Glyugitsir.

level α = 95%, and the coefficient of variation.

the different groups of patients.

Figure 8.

29

100 donors).

serum described by formulas (26) and (27) in the standard CIE method are connected with the transmittance or reflectance spectra and are integrated parameters determined over the entire visible region of the electromagnetic spectrum. So it is assumed that they carry information about the condition of the entire body. In our approach, blood and its components are considered as a single, indivisible light-

New Results in the Theory and Practical Application of Color

DOI: http://dx.doi.org/10.5772/intechopen.84832

absorbing system.

dard procedures [37].

This cycle of works is described [22–26] and executed together with Dr. N. Kalashchenko and Dr. S. Dezortsev. The experiments were conducted at the Ufa Medical University and the Republican Clinic named after Kuvatov (Ufa, Russia).

Colorimetric studies of blood are actively used in medicine [31], criminal law [32], and the food industry [33, 34]. In medical practice, colorimetric methods are used to determine the hemoglobin concentration in the blood of a patient (the color index) [35]. Today a rather exact (�1%) cyanomethemoglobin photometric method is used everywhere, in which cyanomethemoglobin is determined at a wavelength of 540 nm after preparation of a working solution of the blood in Drabkin reagent. Various modifications of this method do not change its essential physical nature [6]. Furthermore, spectral analysis in the visible region has been used to determine oxyhemoglobin and other hemoglobin-containing compounds from the absorption spectra of blood and its solutions [36]. Despite this, the quantitative colorimetric characteristics of blood have not been studied before.

The aim of this work was to study the color characteristics of hemolyzed blood, plasma, and serum from donors in the visible range of the absorption spectra by standard CIE methods (International Commission on Illumination, 1964).

The basic color characteristics (lightness and chromaticity coordinates) determine the position of the color of the specimen in an arbitrary color space and are found by the CIE method [11, 12].

The familiar spectrophotometric method for color measurements involves measuring the spectral power distribution of the radiation followed by calculation of the color coordinates by multiplying the determined spectral power distribution function times the three color-matching functions and then integrating the products. For the spectral power distribution function of the source E(λ), the spectral transmittance function τ(λ), and x(λ), y(λ), and z(λ) (the color-matching functions) and the color coordinates X, Y, and Z are determined by integration over the wavelength range for visible radiation 380–760 nm. In practice, integration is replaced by summation over the interval dλ (from 5 to 10 nm), since the spectral functions under the integral sign are usually not easily integrated:

$$\begin{aligned} X &= \Delta\lambda \sum\_{\lambda} E(\lambda)\tau(\lambda)\overline{\mathbf{x}}(\lambda) \\ Y &= \Delta\lambda \sum\_{\lambda} E(\lambda)\tau(\lambda)\overline{\mathbf{y}}(\lambda), \\ Z &= \Delta\lambda \sum\_{\lambda} E(\lambda)\tau(\lambda)\overline{\mathbf{z}}(\lambda) \end{aligned} \tag{26}$$

The spectral power distribution and the spectral transmittance curve are measured by separating light into a spectrum, such as in a spectrophotometer or monochromator. The color-matching curves are specified as tables of values of the specific coordinates in 10 nm steps. There are also tables of E(λ)x(λ) values for standard CIE light sources A, B, C, and D, characterizing the most typical natural (B, C, D) and artificial (A) illumination conditions.

The chromaticity coordinates are calculated using the formulas.

$$\mathbf{x} = \frac{\mathbf{X}}{\mathbf{X} + \mathbf{Y} + \mathbf{Z}}, \mathbf{y} = \frac{\mathbf{Y}}{\mathbf{X} + \mathbf{Y} + \mathbf{Z}}, \mathbf{z} = \frac{\mathbf{Z}}{\mathbf{X} + \mathbf{Y} + \mathbf{Z}}, \mathbf{x} + \mathbf{y} + \mathbf{z} = \mathbf{1} \tag{27}$$

The coordinate Y characterizes the lightness (luminance) of the specimens.

5. Normal and pathological color characteristics of human blood

This cycle of works is described [22–26] and executed together with Dr. N. Kalashchenko and Dr. S. Dezortsev. The experiments were conducted at the Ufa Medical University and the Republican Clinic named after Kuvatov

characteristics of blood have not been studied before.

under the integral sign are usually not easily integrated:

(B, C, D) and artificial (A) illumination conditions.

<sup>X</sup> <sup>þ</sup> <sup>Y</sup> <sup>þ</sup> <sup>Z</sup> , <sup>y</sup> <sup>¼</sup> <sup>Y</sup>

<sup>x</sup> <sup>¼</sup> <sup>X</sup>

28

X ¼ Δλ ∑ λ

Y ¼ Δλ ∑ λ

Z ¼ Δλ ∑ λ

The chromaticity coordinates are calculated using the formulas.

<sup>X</sup> <sup>þ</sup> <sup>Y</sup> <sup>þ</sup> <sup>Z</sup> , z <sup>¼</sup> <sup>Z</sup>

The coordinate Y characterizes the lightness (luminance) of the specimens.

found by the CIE method [11, 12].

Colorimetric studies of blood are actively used in medicine [31], criminal law [32], and the food industry [33, 34]. In medical practice, colorimetric methods are used to determine the hemoglobin concentration in the blood of a patient (the color index) [35]. Today a rather exact (�1%) cyanomethemoglobin photometric method is used everywhere, in which cyanomethemoglobin is determined at a wavelength of 540 nm after preparation of a working solution of the blood in Drabkin reagent. Various modifications of this method do not change its essential physical nature [6]. Furthermore, spectral analysis in the visible region has been used to determine oxyhemoglobin and other hemoglobin-containing compounds from the absorption spectra of blood and its solutions [36]. Despite this, the quantitative colorimetric

The aim of this work was to study the color characteristics of hemolyzed blood, plasma, and serum from donors in the visible range of the absorption spectra by standard CIE methods (International Commission on Illumination, 1964).

The basic color characteristics (lightness and chromaticity coordinates) determine the position of the color of the specimen in an arbitrary color space and are

The familiar spectrophotometric method for color measurements involves measuring the spectral power distribution of the radiation followed by calculation of the color coordinates by multiplying the determined spectral power distribution function times the three color-matching functions and then integrating the products. For the spectral power distribution function of the source E(λ), the spectral transmittance function τ(λ), and x(λ), y(λ), and z(λ) (the color-matching functions) and the color coordinates X, Y, and Z are determined by integration over the wavelength range for visible radiation 380–760 nm. In practice, integration is replaced by summation over the interval dλ (from 5 to 10 nm), since the spectral functions

Eð Þλ τ λð Þxð Þλ

Eð Þλ τ λð Þyð Þλ ,

(26)

Eð Þλ τ λð Þzð Þλ

<sup>X</sup> <sup>þ</sup> <sup>Y</sup> <sup>þ</sup> <sup>Z</sup> , <sup>x</sup> <sup>þ</sup> <sup>y</sup> <sup>þ</sup> <sup>z</sup> <sup>¼</sup> 1 (27)

The spectral power distribution and the spectral transmittance curve are measured by separating light into a spectrum, such as in a spectrophotometer or monochromator. The color-matching curves are specified as tables of values of the specific coordinates in 10 nm steps. There are also tables of E(λ)x(λ) values for standard CIE light sources A, B, C, and D, characterizing the most typical natural

components

Color Detection

(Ufa, Russia).

The quantitative colorimetric characteristics of hemolyzed blood, plasma, and serum described by formulas (26) and (27) in the standard CIE method are connected with the transmittance or reflectance spectra and are integrated parameters determined over the entire visible region of the electromagnetic spectrum. So it is assumed that they carry information about the condition of the entire body. In our approach, blood and its components are considered as a single, indivisible lightabsorbing system.

The experiment. The objects of investigation were solutions of hemolyzed blood and solutions of plasma and serum (prepared from that blood) of the same concentration from 100 male and female donors (in different blood groups and age groups) and from 95 patients who were assigned to three arbitrary groups: (I) 41 patients with purulent diseases (osteomyelitis, purulent fistulas, gonitis), (II) 41 resuscitated patients (acute myocardial infarction, acute cerebral circulatory collapse, chronic cardiac insufficiency), and (III) 13 patients with cirrhosis of the liver. We determined the color characteristics of the "average" donor (without separating the donors according to blood, sex, and age groups) and compared them with the analogous characteristics of patients from the different groups. The blood for the studies was drawn at a blood donation center and in clinical departments by standard procedures [37].

The spectra of the solutions of hemolyzed blood, plasma, and serum of concentration 2.5 vol.% (1.40) were taken in quartz cuvets with thickness of the working layer of liquid equal to 10 mm at room temperature, on an SF-2000 spectrophotometer in the range 200–1000 nm in 20 nm steps. As the solvent and the reference solution, we used distilled water for injection, which is optically neutral under the experimental conditions and is the natural physiological solvent in the human body. The hemolyzed blood was prepared using a standard heparin solution, and the plasma was prepared using the preservative Glyugitsir.

In addition to the averaged values of the color coordinates and the lightness value, we calculated the standard deviation, the confidence interval for significance level α = 95%, and the coefficient of variation.

Figure 8 shows the averaged spectra for the hemolyzed blood, plasma, and serum from the patients in all three examined groups compared with the corresponding averaged spectra of the donors. There are clear differences between the different groups of patients.

For the plasma and serum, over the entire studied region, the group spectra for the patients lie higher than the averaged spectra of the donors, and their positional order is consistent: the averaged spectrum for patients with purulent diseases lies above the averaged spectrum for the donors, the averaged spectrum for the

### Figure 8.

Spectra of hemolyzed donor blood (1), plasma (2), and serum (3) in the UV and visible regions (averaged over 100 donors).

### Figure 9.

Averaged spectra of hemolyzed blood (a), plasma (b), and serum (c) of examined groups of patients (▲ = donors, ¨ = I, ¤ = II, = III) compared with averaged spectrum of hemolyzed blood, plasma, and serum, respectively, from donors [25].

resuscitation patients lies above that spectrum, and the averaged spectrum for patients with cirrhosis of the liver lies even higher. Probably such positioning of the spectra reflects the severity of the general condition of the patients, if we assume that cirrhosis is the most severe condition for the patients with the least likelihood of recovery. We do not observe such a dependence for the hemolyzed blood: the averaged spectrum for the patients with purulent diseases lies below the averaged spectrum for the donors (Figure 9).

Table 12 gives the averaged color coordinates and lightness for the donors and each group of examined patients, calculated for the solutions of blood, plasma, and serum as a single light-absorbing system according to the standard CIE method. For the donors, the chromaticity coordinate x varies from 0.320 0.001 (for serum and plasma) to 0.630 0.008 (for hemolyzed blood). The coefficients of variation in this case also decrease from 4.7 for blood down to 1.3 for serum. The chromaticity coordinate y for the donors has similar values: 0.320 0.002 for plasma and serum and 0.340 0.003 for blood. The coefficients of variation for y steadily decrease from 3.2 for blood down to 2.0 for serum. The parameter z for the donors is higher for serum and plasma (0.360 0.003) than for blood (0.030 0.005). The coefficient of variation for this parameter is maximum for blood (67.6) and minimum for serum (2.5). The lightness, as expected, has the maximum value (84.88– 1.54) for serum and the minimum value (11.55–0.67) for hemolyzed blood. For plasma, this parameter is close to the value typical of serum.

In determining the color range (see Figure 9a) for the dilute solutions (1.40) of hemolyzed blood, plasma, and serum from the donors, the color range of blood falls within the red region of the spectrum; the range for plasma and serum falls within the yellow region with lower saturation, which supports the correctness of our experiments and calculations. The corresponding regions for the color range for the patients cover a larger area than for the donors (Figure 9b). In order to better visualize the results obtained, we calculated the color coordinates for the studied specimens with correction for concentration. All the points for the donors lie within the yellow-orange region with saturation of 30–50%, which corresponds to the visual observations.

The average values of the color coordinates for all the patient groups (see Table 12) are virtually no different from the averages for the donors except for Parameter

31

Chromaticity

Mean value Standard deviation Confidence interval, α = 0.95

Coefficient of variation

Chromaticity

Mean value Standard deviation Confidence interval, α = 0.95

Coefficient of variation

Chromaticity

Mean value Standard deviation Confidence interval, α = 0.95

Coefficient of variation

Lightness L, %

Mean value Standard deviation Confidence interval, α = 0.95

Coefficient of variation \*Note: I, II, and III indicate the arbitrary groups of patients.

Table 12. Integrated normal and pathological

 color

characteristics

 of hemolyzed

 human blood, plasma, and serum [22–25].

11.55

2.46

 0.67 21.3

 23.86

 21.38

 43.07

 10.3

 18.11

 23.92

 53.6

0.9

 0.85

 2.84

 2.19

3.11

 4.62

 9.63

 1.54

6.6

 13.03

 23.2

 47.35

 2.62

 5.14

 11.15

3.5

 2.72

 5.8

8.17

 12.42

 14.79

 19.65

 5.58

 10.21

 16.46

 22.77

 14.82

 12.99

 13.46

 78.94

 67.82

 63.04

 36.66

 84.88

 79.34

 72.42

 48.08

0.03

0.02

 0.005

67.6

 53.77

 53.16

 61.58

 3.3

4.98

 7.29

 13.46

 2.5

5.53

 7.76

 13.26

 0.007

 0.008

 0.017

 0.003

 0.004

 0.008

 0.019

 0.003

 0.005

 0.008

 0.019

 0.028

 0.024

 0.034

 0.012

 0.017

 0.024

 0.039

 0.009

 0.019

 0.026

 0.038

 0.005

 0.05

 0.06

 0.36

 0.34

 0.33

 0.29

 0.36

 0.35

 0.34

 0.29

 coordinate z

0.34

0.011

 0.003

3.2

3.67

 3.07

 3.98

2.6

3.36

 3.37

 5.27

2.0

3.55

 3.61

 5.22

 0.003

 0.003

 0.007

 0.002

 0.003

 0.004

 0.009

 0.002

 0.003

 0.004

 0.009

 0.013

 0.011

 0.014

 0.008

 0.011

 0.011

 0.019

 0.006

 0.011

 0.012

 0.019

 0.35

 0.35

 0.36

 0.32

 0.33

 0.34

 0.36

 0.32

 0.33

 0.34

 0.37

New Results in the Theory and Practical Application of Color

 coordinate y

0.63

0.029

 0.01

4.7

6.9

 5.46

 8.11

1.6

2.65

 4.13

 5.82

1.3

2.9

 4.93

 5.5

 0.01

 0.01

 0.02

 0.001

 0.002

 0.004

 0.01

 0.001

 0.002

 0.005

 0.009

DOI: http://dx.doi.org/10.5772/intechopen.84832

 0.041

 0.032

 0.048

 0.005

 0.009

 0.013

 0.02

 0.004

 0.009

 0.016

 0.019

 0.59

 0.64

 0.34

 0.32

 0.335

 0.33

 0.35

 0.32

 0.32

 0.32

 0.35

 coordinate x

Hemolyzed blood

Donors

 I

II

III

 Donors

 I

II

III

 Donors

 I

II

III

Plasma

Serum

