Method for determining zinc content in insulin by ion association complex fluorescence quenching

A technology of fluorescence quenching and insulin, applied in measuring devices, fluorescence/phosphorescence, material analysis through optical means, etc., can solve the problems of cumbersome operation, low precision, low detector sensitivity, etc., and achieve strong selectivity and high sensitivity Effect

Inactive Publication Date: 2017-08-29
ANHUI HONGYE PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the commonly used detection method of insulin zinc content is atomic absorption spectrophotometry. However, this method has the disadvantages of cumbersome operation, expensive equipment, long time consumption, low precision and low detector sensitivity, etc., and cannot fully meet the needs.

Method used

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  • Method for determining zinc content in insulin by ion association complex fluorescence quenching
  • Method for determining zinc content in insulin by ion association complex fluorescence quenching
  • Method for determining zinc content in insulin by ion association complex fluorescence quenching

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A method for measuring zinc content in insulin by fluorescence quenching of ion associations, comprising the following steps:

[0023] (1) Preparation of the test solution:

[0024] Weigh 10.0 mg of insulin raw material drug (Tonghua Dongbao, batch number: 161201) in a porcelain crucible, place it on an electric furnace for carbonization for 0.5 h, then place it in a muffle furnace at 500°C for 2 h, take it out and cool it to room temperature, and add 1 mL of 2mol / L Dissolve the residue in HCl solution, transfer it to a 100mL volumetric flask, dilute it with water to the mark and shake it well, and use it as the test solution for later use;

[0025] (2) Zinc standard solution preparation:

[0026] Measure a zinc single element standard solution with a concentration of 1.0mg / mL, and dilute it with 0.01mol / L hydrochloric acid to obtain a 1.0mg / L zinc standard solution;

[0027] (3) Draw a standard curve:

[0028] Draw 0mL, 1mL, 2mL, 3mL, 4mL and 5mL of zinc standard so...

Embodiment 2

[0045] The method for measuring zinc content in insulin injection by fluorescence quenching of ion associates comprises the following steps:

[0046] (1) Preparation of the test solution:

[0047] Measure 10ml of insulin injection (Anhui Hongye Pharmaceutical Co., Ltd., batch number: 161203S): 400 units are placed in a porcelain crucible, evaporated to dryness on an electric furnace, and then carbonized for 0.5h, then ashed in a muffle furnace at 500°C for 2h, taken out and cooled to At room temperature, add 1mL of 2mol / L HCl solution to dissolve the residue, transfer to a 100mL volumetric flask, dilute with water to the mark and shake well, as the test solution for later use;

[0048] Both step (2) and step (3) are the same as step (2) and step (3) of embodiment 1, and will not be repeated here.

[0049] (4) measure the content of zinc in need testing solution:

[0050] In a 25mL volumetric flask, add 5mL of the test solution, add 0.50mL of 1mg / mL potassium iodide solution,...

Embodiment 3

[0054] The method for measuring the zinc content in protamine insulin injection by fluorescence quenching of ion associates comprises the following steps:

[0055] (1) Preparation of the test solution:

[0056] Measure protamine insulin injection (Jiangsu Wanbang Biochemical Medicine, batch number: 161201) 10ml: 400 units in a porcelain crucible, evaporate to dryness on an electric furnace, continue carbonization for 0.5h, then put it in a muffle furnace at 500°C for 2h, take out the cold To room temperature, add 1mL of 2mol / L HCl solution to dissolve the residue, transfer to a 1000mL volumetric flask, dilute with water to the mark and shake well, as the test solution for later use;

[0057] Both step (2) and step (3) are the same as step (2) and step (3) of embodiment 1, and will not be repeated here.

[0058] (4) measure the content of zinc in need testing solution:

[0059] In a 25mL volumetric flask, add 5mL of the test solution, add 0.50mL of 1mg / mL potassium iodide sol...

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Abstract

The invention relates to a method for determining zinc content in insulin by ion association complex fluorescence quenching, which relates to the field of pharmaceutical analysis. The method comprises the following steps: in a volumetric flask, respectively adding a tested object solution or zinc standard solutions with different volumes, respectively adding 0.50 mL of a potassium iodide solution with concentration of 1 mg / mL, placing the material for 5 min, adding a drop of a methyl red indicator, using 0.2 mol / L of an aqueous solution of ammonia for adjusting the color of the solution from red to yellow, respectively adding 2 mL of a phosphatic buffer solution with pH value of 7.2; finally adding 2.0 mL of a rhodamine 6G solution with concentration of 0.025%, adding water for diluting the material to a scale, preparing a to-be-measured solution and a series work solution; scan EEM spectra of the work solution and the to-be-measured solution, recording the fluorescence intensity at 554 nm, drafting a standard curve of fluorescence intensity to zinc concentration, and using an external standard method to calculate the zinc content in the insulin. The method uses a relation that the fluorescence intensity of a rhodamine 6G-potassium iodide system is linearly reduced by zinc ions, so that zinc content in insulin is determined, the operation is simple, the equipment is easy, the detection is rapid and accurate, and the sensitivity is high.

Description

technical field [0001] The invention relates to the field of drug analysis, in particular to a method for measuring zinc content in insulin by fluorescence quenching of ion associates. Background technique [0002] Insulin is the only hormone that lowers blood sugar in the body. The existence of insulin is through two zinc ions (Zn 2+ ) to form a hexameric structure with six insulin monomers. Zinc is related to the synthesis, secretion, storage, hypoglycemic, biological activity and antigenicity of insulin. Zinc deficiency can induce insulin resistance and even diabetes. [0003] As the active component of superoxide dismutase, zinc plays a vital role in the protection of islet cells in the body. It can reduce the inflammatory response of islets through anti-oxidation, inhibit the release of cytokines from macrophages, and reduce the serum levels of diabetic rats. NO and other damage factors, thereby protecting the islet β cells. Zinc can also activate carboxypeptidase B,...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432
Inventor 汪洪源温婷宫传艳陈静吴延芳郭金星
Owner ANHUI HONGYE PHARMA
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