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A kind of detection method of citrulline content in citrulline raw material

A detection method and citrulline technology, applied in the field of biological analysis, can solve the problems affecting the accuracy of quantitative analysis of citrulline, damage to the service life of chromatographic columns, and expensive derivatization reagents, and achieve more convincing and shortened detection results. The effect of detecting time and reducing the error rate of experimental operation

Active Publication Date: 2022-04-22
湖北远大生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The derivation steps of this method are cumbersome and inconvenient to operate
In addition, the derivatization produced many by-products, which made the chromatograms difficult to separate, which in turn affected the accuracy of the quantitative analysis of citrulline
And there are many kinds of derivatization reagents, difficult to screen, expensive derivatization reagents, high detection cost, not suitable for multi-sample detection
[0006] The patent number is CN201810738884.9 (invention name is a high-efficiency liquid phase detection method for L-citrulline), the chromatographic column used is an amino-bonded silica gel column, the detection cost is high, and the mobile phase of this method is used in large quantities Chromatographically pure, the service life of the column is compromised

Method used

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  • A kind of detection method of citrulline content in citrulline raw material
  • A kind of detection method of citrulline content in citrulline raw material
  • A kind of detection method of citrulline content in citrulline raw material

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] The equipment, reagents and detection conditions used in this embodiment are as follows:

[0062] High performance liquid chromatography: Thermo Fisher Dionex Ultimate 3000; VWD UV detector;

[0063] Chromatographic column: Thermo Fisher Syncronis C18 (5μm, 4.6×250mm);

[0064] Column temperature: 30°C;

[0065] Wavelength: 190nm;

[0066] Flow rate: 0.6mL / min;

[0067] Injection volume: 10μL;

[0068] Mobile phase: Mobile phase A is a mixed solution of 0.05mol / L dipotassium hydrogen phosphate and 0.005mol / L sodium decanesulfonate, the pH value is 2.3, and mobile phase B is absolute ethanol;

[0069] Mobile phase volume ratio: 1000:70.

[0070] The specific operation steps for the determination of the content of citrulline in the reference substance:

[0071] (1) Reference substance solution preparation

[0072] Citrulline control stock solution: Accurately weigh an appropriate amount of citrulline reference substance, and dilute with deionized water to obtain a ...

Embodiment 2

[0076] The equipment, reagents and detection conditions used in this embodiment are as follows:

[0077] High performance liquid chromatography: Thermo Fisher Dionex Ultimate 3000; VWD UV detector;

[0078] Chromatographic column: Thermo Fisher Syncronis C18 (5μm, 4.6×250mm);

[0079] Column temperature: 30°C;

[0080] Wavelength: 190nm;

[0081] Flow rate: 0.6mL / min;

[0082] Injection volume: 10μL;

[0083] Mobile phase: Mobile phase A is a mixed solution of 0.05mol / L dipotassium hydrogen phosphate and 0.005mol / L sodium decanesulfonate, the pH value is 2.3, and mobile phase B is absolute ethanol;

[0084] Mobile phase volume ratio: 1000:70.

[0085] Determination of citrulline content in raw materials Concrete steps:

[0086] (1) Take 0.1g of citrulline raw material (the purity of citrulline is known to be 98.11%) and place it in a 100mL volumetric flask, use deionized water to set the volume to the mark, and shake well to obtain citrulline with a concentration of 1g / L...

Embodiment 3

[0090] The equipment, reagents and detection conditions used in this embodiment are as follows:

[0091] High performance liquid chromatography: Thermo Fisher Dionex Ultimate 3000; VWD UV detector;

[0092] Chromatographic column: Thermo Fisher Syncronis C18 (5μm, 4.6×250mm);

[0093] Column temperature: 30°C;

[0094] Wavelength: 190nm;

[0095] Flow rate: 0.6mL / min;

[0096] Injection volume: 10μL;

[0097] Mobile phase: Mobile phase A is a mixed solution of 0.05mol / L dipotassium hydrogen phosphate and 0.005mol / L sodium decanesulfonate, the pH value is 2.3, and mobile phase B is absolute ethanol;

[0098] Mobile phase volume ratio: 1000:70.

[0099] Citrulline linearity test specific steps:

[0100] (1) Accurately draw an appropriate amount of citrulline contrast stock solution prepared in Example 1 for 10 times respectively, place them in five 10ml volumetric flasks respectively, and dilute with deionized water to obtain a concentration of 0.10g / L, 0.20g / L, 0.41g / L, ...

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Abstract

The invention relates to the field of biological analysis, in particular to a method for detecting citrulline content in citrulline raw materials. The invention adopts high-performance liquid chromatography, injects the solution of citrulline raw material into high-performance liquid chromatography, adopts C18 column to separate, and ultraviolet absorption detector to detect. The present invention does not require pre-column or post-column derivatization treatment on the test sample, greatly shortens the test time and makes the test result more accurate. The chromatographic column used in the present invention is an ordinary C18 column, and the detector is an ultraviolet detector, the chromatographic condition is simple, the detection cost is low, the operation is simple and convenient, and it is suitable for large-scale popularization and application.

Description

technical field [0001] The invention relates to the field of biological analysis, in particular to a method for detecting citrulline content in citrulline raw materials. Background technique [0002] Citrulline (Citrulline) is also known as (S)-2-amino-5-ureidovaleric acid or carbamoylornithine, and its molecular formula is C 6 h 13 N 3 o 3 , the molecular weight is 175.19. L-citrulline is a natural non-protein amino acid that is abundant in watermelon. It is an integral part of the urea cycle and plays an important role. When L-citrulline enters the bloodstream, it can be converted to L-arginine and nitric oxide. Nitric oxide can regulate vasodilation, and low L-citrulline content can regulate body fatigue and sexual dysfunction. L-citrulline also keeps the skin healthy and protects the liver. [0003] In recent years, many foreign studies have shown that citrulline also has many important physiological functions, such as scavenging free radicals, stabilizing blood ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/74G01N30/54
CPCG01N30/02G01N30/06G01N30/74G01N30/54G01N2030/027G01N2030/062
Inventor 赵东明刘晓蓉曹亚杰欧阳晖林添雄
Owner 湖北远大生物技术有限公司
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