Calculating molar mass values of components of and molar mass concentration values of conjugate molecules/particles

A method for using an ultraviolet (UV) light signal to analyze a biological solution, comprising:

• separating, by a separation device, a plurality of conjugate molecules of the biological solution having different sizes from each other;
• analyzing, by a light scattering instrument, each of the separated conjugate molecules, wherein for a molecule of the plurality of conjugate molecules, the method further comprises
• receiving, by a computer system, from a differential refractive index (dRI) detector ΔRI values for conjugate molecules/particles with respect to a solvent containing the conjugate molecules/particles,
• wherein the conjugate molecules/particles comprises wavelength absorbing components and non-wavelength absorbing components and are separated according to size by a separation device;
• receiving, by the computer system, wavelength absorption detector ΔUV values for the conjugate molecules/particles, wherein a ΔUV value of the ΔUV values includes a change in a UV extinction at a wavelength of the biological solution relative to a pure solvent due to the molecule, wherein the UV extinction includes an absorbance component and a scattering component;
• receiving, by the computer system, from a light scattering instrument reduced Rayleigh ratio, R(θ), values with respect to the conjugate molecules/particles and the solvent;
• receiving, by the computer system, a dn/dc value for the wavelength absorbing components, dn/dc1, and a dn/dc value for the non-wavelength absorbing components, dn/dc2, from a dn/dc data source;
• receiving, by the computer system, an absorbing extinction coefficient (aEC) value for the wavelength absorbing components, α1, from an aEC data source;
• receiving, by the computer system, a scattering correction factor value for the wavelength absorbing components, D, from a D data source;
• receiving, by the computer system, a fit function, sEC=f(M), for non-wavelength absorbing molecules/particles from a f(M) data source,
• wherein the scattering correction factor value and the fit function provide for the removal of the scattering component of the UV extinction, and
• applying, by the light scattering instrument, a corrected UV signal due to the removal of the scattering component of the UV extinction to analyze the each of the separated conjugate molecules,
• wherein the non-wavelength absorbing molecules/particles comprise the non-wavelength absorbing components;
• and
• executing, by the computer system, a set of logical operations performing conjugate analysis on the ΔRI values, the ΔUV values, the R(θ) values, the dn/dc1 value, the dn/dc2 value, the α1 value, and the D value, with respect to the fit function, sEC=f(M),
• resulting in molar mass values of the wavelength absorbing components, M1, molar mass values of the non-wavelength absorbing components, M2, and molar concentration values of the conjugate molecules/particles.

A system for using an ultraviolet (UV) light signal to analyze a biological solution, comprising:

• a memory;
• and
• a processor in communication with the memory, the processor configured to perform a method comprising
• receiving ΔRI values from a differential refractive index (dRI) detector, ΔUV values from a wavelength absorption (UV) detector, and reduced Rayleigh ratio, R(θ), values from a static light scattering (SLS) instrument, for non-wavelength absorbing molecules/particles, wherein a ΔUV value of the ΔUV values includes a change in a UV extinction at a wavelength of the biological solution relative to a pure solvent due to the molecule, wherein the UV detector separates a plurality of conjugate molecules of the biological solution having different sizes from each other, and the SLS instrument analyzes each of the separated conjugate molecules, wherein the UV extinction includes an absorbance component and a scattering component,
• wherein the non-wavelength absorbing molecules/particles range in size from a lower size limit to an upper size limit and are separated according to size by a separation device, wherein for a molecule of the plurality of conjugate molecules, the processor is configured to perform the method comprising:
• calculating apparent scattering extinction coefficient values of the non-wavelength absorbing molecules/particles, sEC, and molar mass values of the non-wavelength absorbing molecules/particles, M, with respect to the ΔRI values, the ΔUV values, and the R(θ) values,
• executing a set of logical operations fitting the sEC values and the M values to a fit equation,
• resulting in a correlation between the sEC values and the M values,
• resulting in a fit function, sEC=f(M), for the non-wavelength absorbing molecules/particles, wherein the correlation and the fit function provide for the removal of the scattering component of the UV extinction,
• receiving from a second dRI detector second ΔRI values for conjugate molecules/particles with respect to a solvent containing the conjugate molecules/particles,
• wherein the conjugate molecules/particles comprise wavelength absorbing components and non-wavelength absorbing components and are separated according to size by a second separation device,
• receiving from a second wavelength absorption detector second ΔUV values for the conjugate molecules/particles,
• receiving from a second SLS instrument second reduced Rayleigh ratio, R(θ), values with respect to the conjugate molecules/particles and the solvent,
• receiving a dn/dc value for the wavelength absorbing components, dn/dc1, and a dn/dc value for the non-wavelength absorbing components, dn/dc2, from a dn/dc data source,
• receiving an absorbing extinction coefficient (aEC) value for the wavelength absorbing components, α1, from an aEC data source,
• receiving a scattering correction factor value for the wavelength absorbing components, D, from a D data source, and
• executing a set of logical operations performing conjugate analysis on the second ΔRI values, the second ΔUV values, the second R(θ) values, the dn/dc1 value, the dn/dc2 value, the α1 value, and the D value, with respect to the fit function, sEC=f(M),
• resulting in molar mass values of the wavelength absorbing components, M1, molar mass values of the non-wavelength absorbing components, M2, and molar concentration values of the conjugate molecules/particles, and
• providing the scattering correction factor value to the SLS instrument to cause the SLS instrument to apply a corrected UV signal.

A computer program product comprising a computer readable storage medium having program instructions embodied therewith, the program instructions executable by a processor to cause the processor to perform a method comprising:

• separating, by a separation device, a plurality of conjugate molecules of the biological solution having different sizes from each other;
• analyzing, by a light scattering instrument, each of the separated conjugate molecules, wherein for a molecule of the plurality of conjugate molecules, the method further comprises:
• receiving ΔRI values from a differential refractive index (dRI) detector, ΔUV values from a wavelength absorption (UV) detector, wherein a ΔUV value of the ΔUV values includes a change in a UV extinction at a wavelength of the biological solution relative to a pure solvent due to the molecule, and reduced Rayleigh ratio, R(θ), values from a static light scattering (SLS) instrument, for non-wavelength absorbing molecules/particles of the molecules, wherein the UV extinction includes an absorbance component and a scattering component;
• wherein the non-wavelength absorbing molecules/particles range in size from a lower size limit to an upper size limit and are separated according to size by the separation device;
• calculating apparent scattering extinction coefficient values of the non-wavelength absorbing molecules/particles, sEC, and molar mass values of the non-wavelength absorbing molecules/particles, M, with respect to the ΔRI values, the ΔUV values, and the R(θ) values;
• executing a set of logical operations fitting the sEC values and the M values to a fit equation,
• resulting in a correlation between the sEC values and the M values,
• resulting in a fit function, sEC=f(M), for the non-wavelength absorbing molecules/particles;
• receiving from a second dRI detector second ΔRI values for conjugate molecules/particles with respect to a solvent containing the conjugate molecules/particles,
• wherein the conjugate molecules/particles comprise wavelength absorbing components and non-wavelength absorbing components and are separated according to size by a second separation device;
• receiving from a second wavelength absorption detector second ΔUV values for the conjugate molecules/particles;
• receiving from a second SLS instrument second reduced Rayleigh ratio, R(θ), values with respect to the conjugate molecules/particles and the solvent;
• receiving a dn/dc value for the wavelength absorbing components, dn/dc1, and a dn/dc value for the non-wavelength absorbing components, dn/dc2, from a dn/dc data source;
• receiving an absorbing extinction coefficient (aEC) value for the wavelength absorbing components, α1, from an aEC data source;
• receiving a scattering correction factor value for the wavelength absorbing components, D, from a D data source;
• executing a set of logical operations performing conjugate analysis on the second ΔRI values, the second ΔUV values, the second R(θ) values, the dn/dc1 value, the dn/dc2 value, the α1 value, and the D value, with respect to the fit function, sEC=f(M),
• resulting in molar mass values of the wavelength absorbing components, M1, molar mass values of the non-wavelength absorbing components, M2, and molar concentration values of the conjugate molecules/particles, and
• providing the scattering correction factor value to the SLS instrument to cause the SLS instrument to apply a corrected UV signal.