Determination of light absorption pathlength in a vertical-beam photometer
View Patent ↗Photometric methods for determining optical pathlength of liquid samples containing analytes dissolved or suspended in a solvent are provided that rely on determining a relationship between the light absorption properties of the solvent and the optical pathlength of liquid samples containing the solvent. This relationship is used to establish the optical pathlength for samples containing an unknown concentration of analyte but having similar solvent composition. Methods are also provided for determining the concentration of analyte in such samples where both the optical pathlength and the concentration of analyte are unknown. The methods may rely on separately determining, at different wavelengths of light, light absorption by the solvent and light absorption by the analyte. Light absorption by the analyte, together with the optical pathlength so determined, is used to calculate the concentration of the analyte.
1. A method for determining the optical pathlength through a liquid-containing sample comprising:
a. placing the liquid-containing sample in a sample holder which provides a substantially vertical light path through the sample, said vertical light path having an unknown optical pathlength through the sample, and said liquid-containing sample having an unknown absorbance;
b. transmitting light through the sample along the vertical light path, the light comprising a first calibration wavelength and a second calibration wavelength;
the first calibration wavelength and the second calibration wavelength having been preselected within the range of wavelengths of 750-2500 nanometers to provide two characteristic light signal values for the liquid with a predetermined relationship between them and with the vertical optical pathlength through the sample;
c. determining measured light signal values at the first calibration wavelength and at the second calibration wavelength; and
d. determining from the measured light signal values, and the predetermined relationship, the vertical optical light path length through the sample.
2. The method of claim 1 wherein the first calibration wavelength is about 1100 nanometers and the second calibration wavelength is about 1200 nanometers.
3. A method according to claim 1 , wherein the first calibration wavelength and the second calibration wavelength are preselected wavelengths of greater than 1100 nanometers to provide two characteristic light signal values for the liquid with a predetermined relationship between them and with the vertical optical pathlength through the sample.
4. A method according to claim 1 , wherein the light signal values are absorbance or transmittance values.
5. A photometric method for determining an optical pathlength of a sample having an analyte dissolved or suspended in a solvent wherein a relationship between light absorption of the analyte and the optical pathlength of the sample are unknown, the method comprising:
measuring a first light signal resulting from transmission of light, having a first predetermined wavelength, through the sample, wherein the sample has an unknown optical pathlength and absorbance;
measuring a second light signal resulting from transmission of light, having a second predetermined wavelength through the sample; and
determining the optical pathlength of the sample from a predetermined relationship between the first and second light signal, and the optical pathlength of the solvent.
6. The method of claim 5 wherein the first and second wavelength are in a near-infrared region of an electromagnetic spectrum of from 750 nanometers to 2500 nanometers wavelength.
7. The method of claim 5 wherein the predetermined relationship between the first and second light signals and the optical pathlength of the solvent are independent of the temperature of the sample.
8. The method of claim 7 wherein the predetermined relationship between the first and second light signals are measured at temperature isosbestic wavelengths of the solvent.
9. The method of claim 8 wherein one of the isosbestic wavelengths is between 988 and 1008 nanometers.
10. The method of claim 9 wherein a second isosbestic wavelength is between 750 and 930 nanometers.
11. The method of claim 9 wherein a second isosbestic wavelength is between 1050 and 1150 nanometers.
12. A photometric method of determining an optical pathlength of a sample having an analyte dissolved or suspended in a solvent wherein a relationship between light absorption of the analyte and the optical pathlength of the sample are unknown, the method comprising:
measuring a difference between a first and a second light signal resulting from transmission of light of a first and a second predetermined wavelength, respectively, through the sample, wherein the sample has unknown optical pathlengths and absorbances; and
determining the optical pathlength of the sample from a predetermined relationship between the difference and a light absorption pathlength of the solvent.
13. The method of claim 12 wherein the first and second light signals are measured at temperature isosbestic wavelengths of the solvent in a near-infrared region of an electromagnetic spectrum of from 750 nanometers to 2500 nanometers wavelength.
14. The method of claim 13 wherein one of the isosbestic wavelengths is between 988 and 1008 nanometers and the other is between 1050 and 1150 nanometers.
15. The method of claim 13 additionally comprising:
measuring a third light signal resulting from transmission of light of a third predetermined wavelength through the sample; and
determining a ratio of either a fraction of light transmitted by the analyte at the third wavelength, or the absorbance of the analyte at the third wavelength, with respect to the light absorption pathlength of the analyte, by relating the first, second and third light signals to the predetermined relationship.
16. The method of claim 15 wherein the first and second light signals are measured at temperature isosbestic wavelengths of the solvent in a near-infrared region of an electromagnetic spectrum of from 750 nanometers to 2500.
17. The method of claim 15 wherein one of the isosbestic wavelengths is between 988 and 1008 nanometers and the other is between 1050 and 1150 nanometers.
18. The method of claim 13 wherein one of the isosbestic wavelengths is between 988 and 1008 nanometers and the other isosbestic wavelength is between 750 and 930 nanometers.
19. The method of claim 13 wherein one of the isosbestic wavelengths is between 988 and 1008 nanometers and the other isosbestic wavelength is between 750 and 930 nanometers.