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High performance liquid chromatography method for simultaneously detecting contents of melanin and inosinic acid in chicken of silky fowl

A technology of high performance liquid chromatography and melanin, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of complex and time-consuming operation, low recovery rate, and low separation degree of thin-layer chromatography, and achieve shortened detection time, The effect of less sample volume and sufficient sample separation

Active Publication Date: 2013-09-18
HANGZHOU ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ultraviolet spectrophotometry can only achieve rough quantification; capillary electrophoresis also has the problems of low resolution and interference from similar substances such as inosine and adenosine monophosphate; and thin-layer chromatography is complicated, time-consuming, and low in recovery. Has been less used in the determination of inosinic acid

Method used

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  • High performance liquid chromatography method for simultaneously detecting contents of melanin and inosinic acid in chicken of silky fowl
  • High performance liquid chromatography method for simultaneously detecting contents of melanin and inosinic acid in chicken of silky fowl
  • High performance liquid chromatography method for simultaneously detecting contents of melanin and inosinic acid in chicken of silky fowl

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Materials and Methods

[0030] 1. Reagent and solution preparation

[0031] Methanol (chromatographically pure); 30%H 2 o 2 (excellent grade); ethanol, ether, triethylamine, phosphoric acid, hydrochloric acid (analytical grade); 5,6-dihydroxy-2-indolecarboxylic acid (CAS: 352195-40-5, Sigma); inosine acid (CAS: 1953-54-4, Sigma Company); squid melanin (CAS: 8049-97-6, Sigma Company); trypsin, papain (Invitrogen Company).

[0032] 1mol / L potassium carbonate (K2 CO 3 ) solution: 138g K 2 CO 3 , dissolve in water, and dilute to 1000mL.

[0033] 10% sodium sulfite (Na 2 SO 3 ) solution: 10g Na 2 SO 3 , dissolve in water, and dilute to 100mL.

[0034] Phosphoric acid / triethylamine buffer: dissolve 3.5 mL of phosphoric acid and 7.2 mL of triethylamine in 800 mL of water, adjust the pH to 6.5, and dilute to 1000 mL.

[0035] Phosphate (PBS) buffer: 8g NaCl, 0.2g KCl, 1.44g Na 2 HPO 4 and 0.24g KH 2 PO 4 Dissolve in 800 mL of water, adjust the pH of the solution...

Embodiment 2

[0056] Example 2 Precision test

[0057] Draw 20 μL of the mixed standard solution with a concentration of 50 / 5 μg / mL inosinic acid / PTCA, and repeat the injection 6 times. The RSDs of the peak areas of PTCA and inosinic acid were measured to be 0.13% and 0.11%, respectively (n=6), indicating that This method has good precision.

[0058]

Embodiment 3

[0059] Example 3 Stability test

[0060] At 0, 2, 4, 6, 8, and 16 hours, 20 μL of the same test solution was drawn and injected, and within 16 hours, the RSDs of the PTCA and inosinic acid peak areas of the samples were 0.97% and 1.72%, respectively (n=6 ), indicating that the detection value of the test solution is stable within 16 hours.

[0061]

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Abstract

The invention relates to a high performance liquid chromatography method for jointly, rapidly and accurately detecting the contents of melanin and inosinic acid in chicken of silky fowl. The method comprises the following steps of: (1) carrying out protease oscillating enzymolysis on a chicken sample for 24 hours, centrifuging to obtain liquid supernatant which serves as inosinic acid to-be-detected solution; (2) orderly washing precipitate which is subjected to enzymolysis and centrifugation with diethyl ether, absolute ethyl alcohol and ultrapure water, removing lipid and impurities, so as to obtain a melanin crude product, oxidizing the crude product into 2, 3, 5-tricarboxylic acid pyrrole under alkaline condition, extracting with diethyl ether, and drying; and (3) dissolving the dry crystal with the to-be-detected inosinic acid solution, loading and detecting the sample on a high performance liquid chromatograph, and quantifying by an external standard method. The high performance liquid chromatography method only needs little sample to simultaneously detect the contents of the melanin and inosinic acid in the chicken sample of silky fowl, moreover, the method is simple in experimental operation steps, saves detection cost, and is beneficial for acceleration of screening excellent silky fowl breed (series) or combination with both outstanding functions and flavor substances.

Description

technical field [0001] The invention relates to a high-performance liquid chromatography method for combined, rapid and accurate detection of melanin and inosinic acid in black-bone chicken meat, belonging to the field of agricultural analytical chemistry. Background technique [0002] my country is a big chicken raising country in the world. According to FAO statistics, in 2011, the number of chickens raised in the world was 19.94 billion, and the number of chickens raised in China reached 4.61 billion, accounting for 23.1%. Based on differences in regions and customs, local chicken breeds of various shapes are produced by folk selection, such as Beijing oil chicken, Huiyang bearded chicken, Qingyuan Ma chicken, Taihe silky chicken, etc., which have become precious inheritance left to us by our ancestors. resource. Among them, black-bone chicken is also called black-bone chicken, which has the characteristics of black skin, black bone, and black meat. Replenishes deficien...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 章学东张乐王欢欢张成先李庆海陈贤惠楼立峰
Owner HANGZHOU ACAD OF AGRI SCI
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