A high-performance liquid chromatography method for simultaneous detection of melanin and inosinic acid content in black-bone chicken
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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: 2014-10-01
HANGZHOU ACAD OF AGRI SCI
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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
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[0032] 1mol / L potassium carbonate (K2 CO 3 ) solution: 138g K 2 CO 3 , dissolved in water, and adjusted to 1000mL.
[0033] 10% sodium sulfite (Na 2 SO 3 ) solution: 10g Na 2 SO 3 , dissolved in water, and adjusted to 100mL.
[0034] Phosphoric acid / triethylamine buffer solution: dissolve 3.5mL phosphoric acid and 7.2mL triethylamine in 800mL water, adjust the pH to 6.5, and dilute to 1000mL.
[0035] Phosphate (PBS) buffer: 8g NaCl, 0.2g KCl, 1.44g Na 2 HPO 4 and 0.24g KH 2 PO 4 Dissolve in 800mL of water, adjust the pH value of the s...
Embodiment 2
[0056] Embodiment 2 precision test
[0057] Draw 20 μL of inosinic acid / PTCA mixed standard solution with a concentration of 50 / 5 μg / mL, repeat the injection 6 times, and measure the RSDs of the peak areas of PTCA and inosinic acid to be 0.13% and 0.11% respectively (n=6), indicating that The method has good precision.
[0058]
Embodiment 3
[0059] Embodiment 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 the RSDs of the peak areas of PTCA and inosinic acid were 0.97% and 1.72% (n=6) respectively within 16 hours. ), 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...
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