发明名称 Method for Quantitatively Measuring the Concentration of a Compound of Unknown Concentration in Solution
摘要 The present invention relates to methods of spectrophotometric determination of the presence of particular substances in a complex mixture of compounds. This invention relates to methods for determining the concentration of various components of complex biological samples including the determination of the concentration of polyoxyethylene containing compounds including polysorbate in protein containing samples as well as determining the ratio of drug to antibody ratios in antibody-drug conjugate solutions
申请公布号 US2016011099(A1) 申请公布日期 2016.01.14
申请号 US201514545931 申请日期 2015.07.09
申请人 Shih I-Tsung 发明人 Shih I-Tsung
分类号 G01N21/31;G01N21/25 主分类号 G01N21/31
代理机构 代理人
主权项 1. A method for determining the concentration of a second compound in a solution comprising buffer and a first compound and a second compound comprising: a. collecting a set of optical density versus wavelength spectra at two or more path lengths of the buffer of a solution containing first compound only to produce a first buffer background spectra; b. collecting a set of optical density versus wavelength spectra at two or more path lengths for the solution containing first compound only; c. correcting the solution containing first compound spectra with the first buffer background spectra to produce a first compound background corrected spectra; d. selecting two or more data points at a first wavelength from the first compound background corrected spectra and performing linear regression on the data points to produce a first slope; e. selecting two or more data points at a second wavelength from the first compound background corrected spectra and performing linear regression on the data point to produce a second slope; f. calculating the ratio of the slopes at the first and second wavelengths for the first compound; g. collecting a set of optical density versus wavelength spectra at two or more path lengths of a buffer of a solution containing a known concentration of the second compound only to produce a second buffer background spectra; h. collecting a set of optical density versus wavelength spectra at two or more path lengths of a solution containing a known concentration of the second compound only; i. correcting the known concentration of the second compound solution spectra with the second buffer background spectra to produce a known concentration of the second compound background corrected spectra; j. selecting two or more data points at the first wavelength from the known concentration of the second compound background corrected spectra and performing linear regression on the data points to produce a third slope; k. determining the extinction coefficient of the second compound by dividing the third slope by the second compound concentration; l. collecting a set of optical density versus wavelength spectra at two or more path lengths of a buffer of a solution with unknown concentrations of the first and second compounds to produce a third buffer background spectra; m. collecting a set of optical density versus wavelength spectra at two or more path lengths for solution with unknown concentrations of the first and second compounds; n. correcting the solution with unknown concentrations of the first and second compounds spectra with the third buffer background spectra to produce a background corrected spectra for a solution with unknown concentration of the first and second compounds; o. selecting two or more data points at a first wavelength from background corrected spectra of solution with unknown concentrations of the first and second compounds and performing linear regression on the data point to produce a fourth slope; p. selecting two or more data points at a second wavelength from the background corrected spectra of solution with unknown concentrations of the first compound and second compounds and performing linear regression on the data points to produce a fifth slope; q. calculating the slope at the first wavelength for the first compound in the solution with unknown concentrations of the first and second compounds by multiplying the fifth slope to the ratio of the slopes at the first and second wavelengths of the first compound; r. determining the slope value at the first wavelength of the second compound in the solution with unknown concentrations of the first and second compounds by subtracting the slope at the first wavelength of the first compound in the solution with unknown concentrations of the first and second compounds from the fourth slope; and s. determining the concentration of the second compound in the solution with unknown concentrations of the first and second compounds by dividing the slope value of the second compound in the solution with unknown concentrations of the first and second compounds by the extinction coefficient at the first wavelength of the second compound.
地址 Basking Ridge NJ US