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Method for measuring content of metallic ion in metal protein

A determination method and metalloprotein technology, which is applied in the preparation of test samples and the measurement of color/spectral properties, etc., can solve the problems of low repeatability and accuracy, increase the development cost of scientific research projects, and vary widely in analysis results. The effect of improving precision and reproducibility, shortening nebulization processing time, and improving analytical efficiency

Active Publication Date: 2012-03-28
BEIJING TIDE PHARMA
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Problems solved by technology

Since the instruments for analyzing the test samples are currently mainly atomic absorption spectrometers and arc light source emission spectrometers, the time for the samples to be atomized in the instruments is relatively long, and it cannot be guaranteed that the ions in the metalloproteins will fall off completely, and the incomplete falloff of metal ions will cause The repeatability and accuracy of each sample analysis are relatively low; due to the difference in nebulization ability between different instruments, the analysis results of the same sample in different models, or even the same model of instruments, are very different; the samples directly injected The process of metal ion falling off during atomization in the atomizer and burning lamp holder takes a long time, consumes a lot of gaseous mobile phase, and accelerates the loss of precision components in the instrument, which brings inconvenience to research and development and increases the cost of scientific research projects. Development costs

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  • Method for measuring content of metallic ion in metal protein
  • Method for measuring content of metallic ion in metal protein
  • Method for measuring content of metallic ion in metal protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: Determination of Cu ion content in Cu / Zn-SOD

[0025] Experimental Materials Instrument material Cu standard Atomic Absorption Spectrometer AA-400 nitric acid air compressor ddH2O Acetylene cylinder gas line Cu lamp

[0026] 【experimental method】

[0027] (1) Linearity and range

[0028] Prepare blank control solution (1mM dilute nitric acid solution) and 1μg / ml, 2μg / ml, 3μg / ml, 4μg / ml, 5μg / ml Cu standard solutions, measure their absorbance, and draw the first-order relationship between absorbance and Cu standard solution concentration Curves, to evaluate linearity.

[0029] Test results: as shown in Table 1.

[0030] Cu concentration (μg / ml) 0 1 2 3 4 5 Absorbance 0 0.0275 0.0547 0.0812 0.1080 0.1349

[0031] Cu standard solution concentration and absorbance form a first-order linearity, r≥0.99, and the linearity meets the requirements;

[0032] (2) Accuracy

[00...

Embodiment 2

[0051] Embodiment 2: Determination of Zn ion content in Cu / Zn-SOD

[0052] Experimental Materials Instrument material Zn standard Atomic Absorption Spectrometer AA-400 hydrochloric acid air compressor ddH2O Acetylene cylinder gas line Zn lamp

[0053] (1) Linearity and range

[0054]Prepare blank control solution (1mM dilute hydrochloric acid solution) and 0.2μg / ml, 0.4μg / ml, 0.6μg / ml, 0.8μg / ml, 1.0μg / ml Zn standard solutions, measure their absorbance, and draw the absorbance and Zn standard The linearity of the first-order curve of the liquid concentration is evaluated.

[0055] Test results: as shown in Table 4.

[0056] Zn concentration (μg / ml) 0 0.2 0.4 0.6 0.8 1.0 Absorbance 0 0.0243 0.0468 0.0693 0.0937 0.1160

[0057] Zn standard solution concentration and absorbance form a first-order linearity, r≥0.99, and the linearity meets the requirements;

[0058] (2) Accuracy

[0059] Prepar...

Embodiment 3

[0099] Embodiment 3: the mensuration of Fe ion content in hemoglobin

[0100] Experimental Materials Instrument material Fe standard Atomic Absorption Spectrometer AA-400 Nitrous acid: trifluoroacetic acid = 1:1 air compressor ddH2O Acetylene cylinder gas line Fe Hollow Cathode Lamp

[0101] Determination of sample solution:

[0102] Prepare a 1 mM mixed acid solution (pH 5) at a ratio of nitrous acid: trifluoroacetic acid = 1:1. Accurately measure 1ml of Fe standard solution, add 1mM to make up to 10ml, as A solution (100ppm), take A solution and add 1mM mixed acid to dilute, and prepare 5ppm, 4ppm, 3ppm, 2ppm, 1ppm standard solutions respectively. Precisely measure 100μl (20mg / ml) of the hemoglobin aqueous solution to be tested, add 1mM mixed acid to make up to 10ml, and prepare three samples in parallel. Use the atomic absorption spectrometer AAS system to measure, turn on the corresponding Fe hollow cathode lamp, w...

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Abstract

The invention discloses a method for measuring the content of a metallic ion in a metal protein, which comprises the following steps of: pretreating a sample, preparing or treating the sample with an acid material and enabling the pH value of a sample solution to be smaller than 5; and then measuring by a metallic element measuring spectrum (an atomic absorption spectroscopy or an atomic emission spectrometer). According to the method, the problems of low accuracy, poor precision, poor reproducibility, poor specificity and the like in the method for measuring the content of the metallic ion in the metal protein are solved; the detection time can be shortened; the analysis efficiency is improved and the analysis cost is reduced.

Description

technical field [0001] The invention relates to a method for determining the content of metal ions in metalloproteins, in particular to a method for accurately quantifying the content of metal ions in metalloproteins through sample pretreatment. Background technique [0002] Metalloproteins are formed by binding proteins and metal ions. Most of the metal ions are only connected to the protein; a few are connected to a smaller molecule in addition to the protein, such as iron (II) in hemoglobin, in addition to being connected to the protein, it is also connected to the porphyrin ring. Metalloproteins have important physiological functions. For example, hemoglobin is necessary for transporting oxygen; ceruloplasmin can catalyze the oxidation of iron (II) to facilitate the combination of iron (III) and protein to form transferrin, which is used to transport iron, etc. . The content of metal ions in metalloproteins plays a vital role in the function of organisms. Recent studie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N1/28
Inventor 刘红星张扬刘利波齐连权贾慧刘玉静周丽莹肖萱
Owner BEIJING TIDE PHARMA
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