Systems and methods for modulating sensitivity of a photometer
A method for calibrating sensitivity of a photometer includes measuring, by a double-beam spectrophotometer, an absorbance spectrum of a control solution, which has been diluted and includes a control substance. The method further includes linearly regressing the absorbance spectrum of the control solution over a predetermined range of wavelengths and determining whether a first slope of the linearly regressed absorbance spectrum of the control solution falls within a range of slopes of lines obtained from linearly regressing absorbance spectra of a plurality of reference solutions over the predetermined range of wavelengths. A concentration of chromophore in each reference solution is known and the absorbance spectra of the plurality of reference solutions have been obtained by the double-beam spectrophotometer.
1. A method for calibrating sensitivity of a photometer, the method comprising:
measuring, by a double-beam spectrophotometer, an absorbance spectrum of a control solution, which has been diluted and includes a control substance;
linearly regressing the absorbance spectrum of the control solution over a predetermined range of wavelengths;
determining whether a first slope of the linearly regressed absorbance spectrum of the control solution falls within a range of slopes of lines, wherein the lines have been obtained from linearly regressing absorbance spectra of a plurality of reference solutions over the predetermined range of wavelengths, and wherein a concentration of chromophore in each reference solution is known and the absorbance spectra of the plurality of reference solutions have been obtained by the double-beam spectrophotometer; and
in a case where it is determined that the first slope falls within the range of the slopes, calculating a concentration of the chromophore in a reference solution that provides an absorbance spectrum whose slope is substantially equal to the absorbance slope of the control solution over the predetermined range of wavelengths.
2. The method according to claim 1 , wherein the plurality of reference solutions are blood samples, which have been diluted.
3. The method according to claim 1 , wherein the chromophore in the plurality of reference solutions is oxygenated hemoglobin.
4. The method according to claim 1 , wherein the control substance is a dye.
5. The method according to claim 4 , wherein the dye is one of malachite green, patent blue V, indigo carmine, brilliant blue, or crystal violet.
6. The method according to claim 4 , wherein an absorbance spectrum of the control substance has a profile shape different from a profile shape of the absorbance spectrum of the chromophore in the plurality of reference solutions.
7. The method according to claim 6 , wherein, in a case where it is determined that the first slope does not fall within the range of the slopes, the method further includes:
Adjusting the concentration of the control substance in the control solution or reformulating the control solution to a target slope of the absorbance spectrum obtained at the adjusted concentration to be substantially equal to a slope within the range of slopes of the plurality of reference solutions over the predetermined range of wavelengths.
8. The method according to claim 1 , wherein the concentration of the chromophore in each of the plurality of reference solutions is different.
9. The method according to claim 1 , wherein the plurality of reference solutions and the control solution are diluted at a same dilution ratio prior to measuring the absorbance spectrum thereof.
10. The method according to claim 1 , wherein the predetermined range of wavelengths is from 564 nanometers (nm) to 576 nm.
11. The method according to claim 1 , wherein an absorbance spectrum of each reference solution, which has been diluted, is accepted when coefficients of variation of absorbances determined at each wavelength for each reference solution, which has been diluted, are lower than a first predetermined threshold.
12. The method according to claim 1 , wherein the lines determined in the linear regression of spectrophotometric absorbances at each wavelength for each reference solution, which has been diluted, are accepted when a coefficient of determination of each line thereof is greater than or equal to a second predetermined threshold.
13. The method according to claim 12 , wherein the absorbance spectrum of the control solution, which has been diluted, is accepted when coefficients of variation of the absorbances determined at each wavelength for the control solution, which has been diluted, are lower than a third predetermined threshold.
14. The method according to claim 1 , wherein the line determined in the linear regression of the spectrophotometric absorbances for each wavelength within the predetermined range for the control solution, which has been diluted, is accepted when a coefficient of determination thereof is greater than or equal to a fourth predetermined threshold.
15. A method for calculating a concentration of a chromophore in a target solution, in which concentration of the chromophore is unknown, by a photometer, the method comprising:
setting a standard concentration of a chromophore in a standard solution over a predetermined range of wavelengths, where an absorbance spectrum of the standard solution has a slope substantially equal to a slope of an absorbance spectrum of a control solution over the predetermined range of wavelengths;
measuring, by a photometer, an absorbance of the control solution;
determining an expected photometric absorbance of the standard solution based on the photometric absorbance of the control solution and the standard concentration;
measuring a photometric absorbance of the target solution; and
calculating a concentration of a chromophore in the target solution based on the photometric absorbance of the target solution, the expected photometric absorbance of the standard solution, and the standard concentration of the standard solution.
16. The method according to claim 15 , wherein the target solution is diluted blood.
17. The method according to claim 15 , wherein the chromophore is oxygenated hemoglobin.
18. The method according to claim 15 , wherein a dominant wavelength of a light source of the photometer falls within the predetermined range.
19. The method according to claim 15 , wherein the standard concentration is set by a linear interpolation based on slopes of absorbance spectra of a plurality of reference solutions and the slope of the absorbance spectrum of the control solution.
20. The method according to claim 15 , wherein the standard solution, the control solution, and the target solution are diluted with a same diluent.
21. The method according to claim 20 , wherein the diluted control solution is diluted at a dilution ratio of the diluted standard solution.
22. The method according to claim 21 , wherein the diluted target solution is diluted at the dilution ratio of the diluted standard solution.
23. The method according to claim 15 , where the expected photometric absorbance of the standard concentration of the standard solution A expected is determined by the following equation:
A
expected
=
A
control
+
υ
·
1
photo
1
spectr
,
where A control is the measured photometric absorbance of the control solution, ν is a constant absorbance offset, l photo is an optical pathlength of a container used for the target solution by the photometer, and l spectr is an optical pathlength of a container used in a double-beam spectrophotometer.
24. The method according to claim 15 , wherein the photometric absorbance of the control solution is accepted when a coefficient of variation thereof is less than or equal to a predetermined threshold.
25. The method according to claim 15 , wherein the photometric absorbance of either of the control or target solution is measured by the following equation:
A
=
log
10
(
I
0
I
)
,
where A is the photometric absorbance of the control or target solution, I 0 is a light intensity detected passing through a container, which is filled with diluent, for the control or target solution, and I is a light intensity detected passing through the container, which holds the control or target solution.
26. The method according to claim 15 , wherein the concentration of the chromophore in the target solution is calculated by the following equation:
c
t
=
A
t
·
(
c
s
A
s
)
,
where c t is the concentration of the chromophore in the target solution, A t is the measured photometric absorbance of the target solution, c s is the standard concentration of the chromophore in the standard solution, and A s is the expected photometric absorbance of the chromophore in the standard solution.