Method for determining cattle lactalbumin content in milk powder and milk by ultra performance liquid chromatography
An ultra-high performance liquid phase, albumin technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problem of high price of mass spectrometers, and achieve the effect of removing impurity interference, improving sensitivity, and ensuring quantitative accuracy
Active Publication Date: 2013-02-13
WISSUN INT NUTRITION GRP
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Problems solved by technology
Due to the high price of the mass spectrometer with a mass-to-nuclei ratio of 3000 used in the national standard method, and the limitations of gel permeation chromatography detection
Method used
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[0032] The present invention is described in detail below in conjunction with embodiment
[0033] experimental method
[0034] Materials, Reagents and Instruments
[0035] Acetonitrile (chromatographically pure), trifluoroacetic acid (excellent pure), Triton X-100, sodium chloride (excellent pure), hydrochloric acid (excellent pure), sodium hydroxide (analytical pure), and experimental water is ultrapure water (18 MΩ cm, TOC 3 ppb), bovine whey protein standards Bα-La, Bβb-Lg and Bβa-Lg; ACQUITY UPLC? ultra-high performance liquid chromatography system, equipped with Acquity TUV detector, FILTER MIXER (425 μL, P / N 205000403).
[0036] test reagent
[0037] 1. 0.1% trifluoroacetic acid in acetonitrile solution: draw 1 mL of trifluoroacetic acid and dilute to 1000 mL with acetonitrile;
[0038] 2. 0.1% trifluoroacetic acid aqueous solution: absorb 1 mL of trifluoroacetic acid and dilute to 1000 mL with water;
[0039]3. 0.1 mol / L hydrochloric acid: absorb 9 mL hydrochloric ...
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Abstract
The invention discloses a method for determining cattle lactalbumin content in milk powder and milk by an ultra performance liquid chromatography. The method is characterized by comprising the following steps of: adding a hydrochloric acid extracting solution containing TritonX-100 into the milk powder and milk to be detected and dissolving by using ultrasound, adjusting pH to be 4.4 to 4.6 by using a sodium acetate aqueous solution, metering volume and homogenizing by using water, standing at room temperature and then filtering, allowing filtrate to pass through a filtration membrane, and putting on an ACQUITYUPLCBEH300C18 chromatographic column, and determining the content of three kinds of cattle lactalbumin, namely Ba-Lalpha, Bbetab-Lg and Bbetaa-Lg in the milk powder and the milk by an ACQUITYUPLC-TUVCQUITYUPLCBEH300C18 chromatographic column. Relatively simple and efficient purification steps are applied to operation, so that the separation of Ba-Lalpha, Bbetab-Lg and Bbetaa-Lg is realized, quantification accuracy is guaranteed, the interference of impurities such as casein, lipid and the like can be effectively removed, and the final detection flexibility is improved. The detection limit of the method is 1 mu g / g.
Description
technical field [0001] The invention belongs to a method for measuring bovine whey protein content in milk powder and milk by ultra-high performance liquid chromatography. Background technique [0002] Generally speaking, protein that has the complete types and contents of essential amino acids and can provide the body's needs can be called high-quality protein, also called complete protein. Essential amino acids refer to amino acids that are necessary for the human body but cannot be synthesized by themselves and must be ingested from food. Among plant proteins, only soy protein is a complete protein, but soy protein is not as absorbable as high-quality animal protein. Whey protein is a high-quality complete protein, and it is also animal protein. It contains 8 kinds of amino acids necessary for the human body, and the ratio is reasonable, close to the proportion required by the human body. It is an indispensable essence for human growth, development, anti-aging and other...
Claims
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IPC IPC(8): G01N30/02G01N30/06G01N30/60
Inventor 陈光耀林玉宙
Owner WISSUN INT NUTRITION GRP