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A method for rapid detection of hydroxyproline content in milk and dairy products

A technology for hydroxyproline and dairy products, applied in the field of analytical chemistry, can solve the problems of easy occurrence of false positives, complicated operation, long detection time, etc., and achieve the effects of simple and fast sample processing, high method sensitivity, and accurate and reliable results.

Active Publication Date: 2018-08-17
INSPECTION & QUARANTINE TECH CENT OF NINGBO ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the colorimetric method is the most commonly used method because of its simple operation, economical and practical, but due to the lack of corresponding separation means, it is prone to false positives
In addition, since most amino acids have no UV absorption, liquid chromatography and amino acid analyzers need to be derivatized before they can be detected. The whole operation is complicated, the detection time is long, and it is easy to pollute the environment

Method used

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  • A method for rapid detection of hydroxyproline content in milk and dairy products
  • A method for rapid detection of hydroxyproline content in milk and dairy products
  • A method for rapid detection of hydroxyproline content in milk and dairy products

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] 1.1 The reagents, instruments and consumables used in the implementation are as follows:

[0032] Thermo ICS-5000 ion chromatograph (including DP quaternary gradient pump and SP single pump, DC control unit, EG eluent generator, AS autosampler), Chromeleon 6.8 chromatographic workstation, Elix Advantage pure water system (U.S. Libo Company), Milli-Q Direct ultrapure water system (Millipore Company of the United States), DGG-9053AD electric heating constant temperature blast drying oven (Shanghai Senxin Experimental Instrument Co., Ltd.), HH-2 digital display constant temperature water bath (Guohua Electric Co., Ltd.), 723PCS Visible Spectrophotometer (Shanghai Xinmao Instrument Co., Ltd.), TOPEX All-round Microwave Chemistry Work Platform (Shanghai Yiyao Instrument Technology Development Co., Ltd.), Vortex-Genie 2 Vortex Mixer (American Siboming Scientific Equipment Co., Ltd. ), SK8210HP ultrasonic cleaner (Shanghai Kedao Ultrasonic Instrument Co., Ltd.), SARTORIOUS QUI...

Embodiment 2

[0066] Use the microwave hydrolysis method in step 2.1.2 as the sample pretreatment method, but change the hydrolysis temperature to 120°C, 140°C, 145°C, and 150°C, and use step 2.2.1 colorimetry and 2.2.2 ion chromatography to detect The content of L-hydroxyproline content in the hydrolyzate, the results are shown in Table 3.

[0067] table 3

[0068]

[0069] the result shows,

[0070] a. When the hydrolysis temperature is between 120 and 150°C, the calculation result is low due to incomplete hydrolysis of the sample.

[0071] b. Among them, in the hydrolysis group at 120°C, the values ​​of the two determination methods differ greatly, because for the colorimetric method, the concentration is less than its quantification limit, and the measurement value is inaccurate; while for the ion chromatography method, the concentration Greater than its limit of quantification, the measured value is reliable.

[0072] c. Except for the 120°C hydrolysis group, in the other tempera...

Embodiment 3

[0074] The milk powder sample was pretreated according to step 2.1.1 of Example 1, traditional acid hydrolysis method and 2.1.2 microwave acid hydrolysis method, except that the added 500 μL of 15 g / L gelatin solution was replaced by 250, 500, and 1000 μg respectively The L-hydroxyproline, and then the obtained hydrolyzate is detected by the colorimetric method of embodiment 1 step 2.2.1, and parallelized for 6 times, the recovery and precision of hydroxyproline are calculated, and the results are as follows Table 4 shows.

[0075] Table 4

[0076]

[0077]

[0078] The recovery calculation formula is as follows (the negative sample does not contain hydroxyproline):

[0079]

[0080] c1 is the concentration (μg / mL) of hydroxyproline in the hydrolyzate of determination;

[0081] n is the dilution factor, where the colorimetric method is 10, and the ion chromatography is 100;

[0082] m1 is the mass of hydroxyproline added (mg).

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Abstract

The invention discloses a method for rapidly detecting the hydroxy proline content of milk and dairy products. A sample is pre-processed by a microwave hydrolysis method. Then the preprocessed sample is subjected to chromatographic column analysis by adopting ion chromatography to obtain the content of L-hydroxy proline. The provided method has the advantages that the sample processing is rapid and simple, the result is accurate and reliable, the sensitivity is better compared with that of national standard method, the content of L-hydroxy proline in milk and dairy products, which contain animal protein hydrolysate, can be accurately measured, the animal protein hydrolysate can be quantitatively and qualitatively analyzed, and a reliable detection method is provided for preventing adulteration.

Description

technical field [0001] The invention relates to the technical field of analytical chemistry, in particular to a method for rapidly detecting the content of hydroxyproline in milk and dairy products. Background technique [0002] The Ministry of Health announced in February 2009 the "List of Non-edible Substances That May Be Added Illegally in Food (Second Batch)" that clearly stipulates that the addition of leather hydrolyzed protein is prohibited in milk, dairy products, and milk-containing beverages. The main protein type of leather hydrolyzed protein is collagen, and hydroxyproline is a unique amino acid in collagen, accounting for about 13.63% of the total collagen amino acids. Milk protein does not contain hydroxyproline, so hydroxyproline is often used as evidence of adulteration of milk powder and milk beverage products. [0003] There are many methods for the determination of hydroxyproline, and the common methods of hydrolyzing protein in the pretreatment process i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/027G01N2030/067
Inventor 葛晓鸣葛一超钟莺莺计红俞雪钧
Owner INSPECTION & QUARANTINE TECH CENT OF NINGBO ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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