Detection on content of lincomycin in feeds through liquid chromatography-mass spectrography method

Abstract

The invention discloses a method of detecting the content of lincomycin hydrochloride in a feed through a high performance liquid chromatography method. A waters 2695 high performance liquid chromatograph is employed with a C18 chromatographic column; a mobile phase includes a 0.05 mol / L borax solution and methanol according to ratio of 55:45; flow rate is 1.0 ml / min; detection wavelength is 214 nm. The method has good separation between a main peak and adjacent peaks, in the range of 14.7-492 [mu]g / ml, the lincomycin hydrochloride can form good linearity, R2 = 0.9987 (n=6). The method is quick, sensitive and reliable, can be used for content detection of the lincomycin hydrochloride in a feed additive. The LC-MS / MS method can supply structural information of a to-be-detected substance; the method is high in sensitivity and selectivity and is an excellent method of detecting various antibiotics. The method is quick, sensitive, reliable and accurate.

Description

technical field

[0001] The invention belongs to the technical field of food safety detection, and in particular relates to a liquid chromatography-mass spectrometry / mass spectrometry method for detecting the content of lincomycin in feed. Background technique

[0002] Antibiotics are not only used as drugs to prevent and treat bacterial infectious diseases in the livestock and poultry industry, but also as feed additives to promote animal growth and improve economic efficiency, so they are widely used in animal feed. However, in recent years, food safety incidents caused by feed have occurred continuously. After a large amount of antibiotics are ingested into the body, they will be distributed in lymph nodes, kidneys, liver and other organs along with the blood circulation, which will reduce the immunity of livestock and poultry, and pathogenic bacteria will take advantage of it and cause more serious diseases. Serious harm, in addition, long-term excessive or improper use o...

Examples

Embodiment 1

[0029] The assay method of embodiment 1 liquid chromatography erythromycin, lincomycin

[0030] 1. When acidified methanol solution was used as the extraction solvent, the recoveries of lincomycin and erythromycin were all less than 10%, which may be due to the glycoside bond structure in lincomycin and erythromycin under acidic conditions. Hydrolysis occurs. In this study, methanol extraction was finally used. The recoveries of the two substances were high and the extraction process was simple.

[0031] 2. The addition of saturated lead acetate

[0032] After the supernatants were combined and blown nearly dry with nitrogen, 5 mL of 0.1 mol / L phosphate buffer solution was added to dissolve them. When the extract was to be purified, 100 μL of saturated lead acetate (adsorbed protein, In order to prevent the column from being blocked during purification), vortex and centrifuge to obtain the supernatant to achieve the purpose of removing protein.

[0033] 2. Selection of extr...

Embodiment 2

[0039] Embodiment 2 lincomycin stability

[0040] The standard stock solutions of lincomycin and erythromycin were placed at -18°C, and their stability was investigated at 1 month, 3 months and 6 months respectively. The experimental results showed that at 6 months Within the period, the stock solutions of lincomycin and erythromycin had no degradation phenomenon, so the storage period of the stock solutions of the two analytes was stipulated in the text as 6 months.

[0041]Lincomycin and erythromycin standard solutions with a concentration of 1 μg / mL were injected into the mass spectrometry system with a needle pump. Using Q1 full scan (Q1 MS) and fragment ion scan (Product Ion MS2) to determine precursor ions and product ions respectively, and optimize related parameters, Figures 1 to 4 It is the mass spectrum of 1 μg / mL lincomycin and erythromycin standard solution in Q1 and Product Ion (MS2) mode. Table 1 is the qualifier and quantifier ions of lincomycin and erythromy...

Embodiment 3

[0053] Embodiment 3 lincomycin in liquid chromatograph

[0054] 1. Quantitation limit and detection limit

[0055] Prepare a mixed standard solution of lincomycin and erythromycin, dilute it to a certain concentration of standard solution: 1.0, 2.0, 5.0, 10.0, 20.0, 50.0, 100.0 μg / L, and carry out under the best chromatographic and mass spectrometric conditions Determination, linear regression analysis was carried out with the peak area as the ordinate and the mass concentration as the abscissa. Gradually dilute the mixed standard solution, and take the signal-to-noise ratio greater than 10 (S / N≥10) as the lower limit of determination. The linear equation and correlation coefficient are shown in Table 3; the curve is as follows Figure 6 and 7 shown.

[0056]

[0057] 2. Precision

[0058] Using the addition method, that is, adding aflatoxin B to samples without aflatoxin in the background 1 , G 1 2.5 µg / kg, 5.0 µg / kg, 25 µg / kg, aflatoxin B 2 , G 2 Three levels of ...

technical field

[0001] The invention belongs to the technical field of food safety detection, and in particular relates to a liquid chromatography-mass spectrometry / mass spectrometry method for detecting the content of lincomycin in feed. Background technique

[0002] Antibiotics are not only used as drugs to prevent and treat bacterial infectious diseases in the livestock and poultry industry, but also as feed additives to promote animal growth and improve economic efficiency, so they are widely used in animal feed. However, in recent years, food safety incidents caused by feed have occurred continuously. After a large amount of antibiotics are ingested into the body, they will be distributed in lymph nodes, kidneys, liver and other organs along with the blood circulation, which will reduce the immunity of livestock and poultry, and pathogenic bacteria will take advantage of it and cause more serious diseases. Serious harm, in addition, long-term excessive or improper use o...

Examples

Embodiment 1

[0029] The assay method of embodiment 1 liquid chromatography erythromycin, lincomycin

[0030] 1. When acidified methanol solution was used as the extraction solvent, the recoveries of lincomycin and erythromycin were all less than 10%, which may be due to the glycoside bond structure in lincomycin and erythromycin under acidic conditions. Hydrolysis occurs. In this study, methanol extraction was finally used. The recoveries of the two substances were high and the extraction process was simple.

[0031] 2. The addition of saturated lead acetate

[0032] After the supernatants were combined and blown nearly dry with nitrogen, 5 mL of 0.1 mol / L phosphate buffer solution was added to dissolve them. When the extract was to be purified, 100 μL of saturated lead acetate (adsorbed protein, In order to prevent the column from being blocked during purification), vortex and centrifuge to obtain the supernatant to achieve the purpose of removing protein.

[0033] 2. Selection of extr...

Embodiment 2

[0039] Embodiment 2 lincomycin stability

[0040] The standard stock solutions of lincomycin and erythromycin were placed at -18°C, and their stability was investigated at 1 month, 3 months and 6 months respectively. The experimental results showed that at 6 months Within the period, the stock solutions of lincomycin and erythromycin had no degradation phenomenon, so the storage period of the stock solutions of the two analytes was stipulated in the text as 6 months.

[0041]Lincomycin and erythromycin standard solutions with a concentration of 1 μg / mL were injected into the mass spectrometry system with a needle pump. Using Q1 full scan (Q1 MS) and fragment ion scan (Product Ion MS2) to determine precursor ions and product ions respectively, and optimize related parameters, Figures 1 to 4 It is the mass spectrum of 1 μg / mL lincomycin and erythromycin standard solution in Q1 and Product Ion (MS2) mode. Table 1 is the qualifier and quantifier ions of lincomycin and erythromy...

Embodiment 3

[0053] Embodiment 3 lincomycin in liquid chromatograph

[0054] 1. Quantitation limit and detection limit

[0055] Prepare a mixed standard solution of lincomycin and erythromycin, dilute it to a certain concentration of standard solution: 1.0, 2.0, 5.0, 10.0, 20.0, 50.0, 100.0 μg / L, and carry out under the best chromatographic and mass spectrometric conditions Determination, linear regression analysis was carried out with the peak area as the ordinate and the mass concentration as the abscissa. Gradually dilute the mixed standard solution, and take the signal-to-noise ratio greater than 10 (S / N≥10) as the lower limit of determination. The linear equation and correlation coefficient are shown in Table 3; the curve is as follows Figure 6 and 7 shown.

[0056]

[0057] 2. Precision

[0058] Using the addition method, that is, adding aflatoxin B to samples without aflatoxin in the background 1 , G 1 2.5 µg / kg, 5.0 µg / kg, 25 µg / kg, aflatoxin B 2 , G 2 Three levels of ...

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