GB/T 5009.194-2003 Determination of immunoglobulin IgG in health foods

This standard specifies the determination method of immunoglobulin IgG in health foods. This standard is used for the determination of immunoglobulin IgG in health foods of tablet, capsule and powder type. The detection limit of this method is: when the sample volume is 0.1g, diluted to 25mL, and the injection volume is 20μL, the detection concentration is 0.5mg/mL. GB/T 5009.194-2003 Determination of immunoglobulin IgG in health foods GB/T5009.194-2003 Standard download decompression password: www.bzxz.net

ICS67.040
National Standard of the People's Republic of China
GB/T5009.194--2003
Determination of immunoglobulin IgG in health foods2003-08-11PromulgatedwwW.bzxz.Net
Ministry of Health of the People's Republic of China
National Administration of Standardization of China
2004-01-01Implementation
This standard is proposed and managed by the Ministry of Health of the People's Republic of China. This standard is drafted by the Food Hygiene Supervision and Inspection Institute of the Ministry of Health. The main drafters of this standard are Yang Zuying and Song Shufeng. GB/T5009.194—2003
GB/T5009.194—2003
Immunoglobulin IgG has been added to health foods as an effective ingredient. This ingredient has certain effects on enhancing immunity, regulating the physiological functions of animals and the metabolism of certain specific substances. This method is based on the immunoglobulin IgG determination method of New Zealand Health Food Co., Ltd. and is improved according to the instrument and equipment conditions in my country. A high-performance liquid chromatography determination method for immunoglobulin content in tablet, capsule and powder type health foods is established. 540
Scope of application
Determination of immunoglobulin IgG in health foods
This standard specifies the determination method of immunoglobulin IgG in health foods. This standard is applicable to the determination of immunoglobulin IgG in tablet, capsule and powder type health foods. GB/T5009.194—2003
The detection limit of this method is: when the sample volume is 0.1g, diluted to 25mL, and the injection volume is 20μL, the detection concentration is 0.5mg/mL. 2 Principle
According to the principle of high-performance affinity chromatography, immunoglobulin IgG is connected to the ligand in a phosphate buffer strip, and immunoglobulin IgG is eluted under the conditions of hydrochloric acid and glycine at pH 2.5. 3 Reagents
3.1 Mobile phase A: pH 6.5, 0.05 mol/L phosphate buffer. 3.2 Mobile phase B: pH 2.5, 0.05 mol/L glycine hydrochloride buffer. 3.3 IgG standard stock solution: Weigh 0.0100 g of IgG standard (Sigma Chemical Company), dissolve it in mobile phase A (3.1) and dilute it to 10.0 mL, shake well, and the concentration is 1.0 mg/mL. 3.4 IgG standard series solution: Take IgG standard stock solution and dilute it with mobile phase A (3.1) to a standard series containing IgG 0.20.4, 0.6, 0.81.0 mg/mL. Prepare it when it is used. 4 Instruments and equipment
High performance liquid chromatography with UV detector and gradient elution device. 5 Analysis steps
5.1 Sample treatment: Weigh 0.1g (accurate to 0.001g) sample, dilute to 25.0mL with mobile phase A (3.1), shake well, pass through 0.45um microporous filter membrane and inject.
5.2 First wash the column with 5 column volumes of redistilled water, then balance the column with 10 column volumes of mobile phase A (3.1), inject sample, and elute according to the elution program.
5.3 HPLC reference conditions
Chromatographic column: Pharmacia HI-Trap Protein G column, 1mL. Wavelength: 280nm.
Injection volume: 20μL.
See Table 1 for the gradient elution table.
Flow rate/（mL/min）
GB/T5009.194—2003
Result calculation
Where:
cXVx100
mx1000
—The content of IgG in the sample, in grams per hundred grams (g/100g)X-
The mass of the sample, in grams (g),
The content of IgG in the test liquid, in milligrams per milliliter (mg/mL); VThe volume of the sample, in milliliters (mIL.). The calculation result shall retain two significant figures.
Precision
The absolute difference between two independent determination results obtained under repeatability conditions shall not exceed 10% of the arithmetic mean. 8Chromatogram
IgG peak
Figure 1IgG standard chromatogram
IgG peak
Figure 2Colosin chromatogram
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