What Is Colorimetry?
Colorimetry is a chemical analysis method used for qualitative or quantitative measurement based on the color or color intensity of a solution.
The Core Principle:
The substance of interest reacts with a specific reagent to produce a colored compound. The depth of color (i.e., absorbance) of this compound is directly proportional to the concentration of the target substance under defined conditions (following the Lambert-Beer Law). By measuring its absorbance at a specific wavelength and comparing it to standards of known concentration, the amount of the target substance can be calculated.
Key Components:
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Chromogenic Reaction: Must be highly specific (reacting only with the target) and highly sensitive (producing a distinct color change).
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Measurement Instrument: Typically, a spectrophotometer or microplate reader is used to measure absorbance precisely at a specific wavelength.
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Standard Curve: A curve plotting absorbance against concentration, created using a series of known standards, is fundamental for quantitative analysis.
A Typical Application in IVD (using ALP as an example):
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Substrate: For instance, ALP catalyzes the breakdown of colorless pNPP to produce yellow p-nitrophenol.
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Measurement: The rate of formation or final concentration of the yellow product is monitored at a wavelength of 405 nm.
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Calculation: The change in absorbance is used to calculate the activity or concentration of ALP in the sample.
In simple terms, colorimetry is the method of "measuring quantity by observing color intensity." It offers advantages such as simple operation, low cost, and ease of automation, making it one of the most fundamental and widely used techniques in clinical chemistry testing and immunoassays (like ELISA).
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