Method for determining lincosamine antibiotics in feed
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A technology for antibiotics and lincomycin, applied in the field of determination of lincomycin antibiotics
Active Publication Date: 2022-07-29
ZHEJIANG UNIV
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So far, there is no literature report on simultaneous det
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Embodiment 1
[0032] Example 1 LC-MS / MS analysis conditions
[0033] The mobile phase consists of two phases A and B (A: 0.1% formic acid in water; B: acetonitrile), gradient elution, see Table 1 for the elution procedure; the flow rate is 0.4 mL min -1 ; the column temperature was set to 30°C; the injection volume was 8 µL.
[0034]
[0035]Lincomycin, clindamycin, and piromycin all have tertiary amine structures and are weakly basic. They have a strong positive ion response in mass spectrometry. Therefore, positive ion mode scanning and multi-stage reaction monitoring ( MRM). The atomizing gas and drying gas are high-purity nitrogen, and the collision gas is high-purity argon. Prepare a concentration of 100 μg L with A and B mobile phases (50:50, V / V) -1 3 compounds lincomycin, clindamycin, and piromycin, 10 mL min by syringe pump -1 The flow rate was injected into mass spectrometry to optimize mass spectrometry parameters such as cone voltage, collision energy, dwell time, and fra...
Embodiment 2
[0037] Example 2 Methodological study
[0038] The method of the present invention was validated with reference to European Commission Decision No. 2002 / 657 / EC (2002 / 657 / EC) to evaluate matrix effects, accuracy (recovery), precision (reproducibility), linearity and selectivity, and Minor modifications to it: Limit of detection (LOD) and limit of quantification (LOQ) are used instead of limit of detection determination (CCα) and detection capacity (CCβ) commonly found in animal feed (Frenich et al., 2011; Ying et al., 2013; Kiebooms et al, 2015)
[0039] In the present invention, the matrix effect (ME) is assessed as the ratio of the slope of the working curve of the matrix-matched standard solution to the slope of the working curve of the standard solution of pure solvent (Chawla et al., 2017; Varga et al., 2021). For the preparation of a series of mixed standard working solutions (1.0, 2.0, 5.0, 10.0, 50.0, 100.0 and 200 μg L -1 ). The matrix effect was calculated accordin...
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Abstract
The invention discloses a method for determining lincosamine antibiotics in feed. The method comprises the following steps: extracting three lincosamine antibiotics (lincomycin, clindamycin and pirithromycin) in a feed by adopting a 80% methanol dipotassium phosphate solution, and purifying a sample by adopting a mixed cation exchange (MCX) solid-phase extraction small column. The lincomycin, the clindamycin and the pirithromycin are quantified at the same time through liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results show that under the addition level of 40-4000 [mu] g kg <-1 >, the average recovery rate of lincosamines in seven feeds is 81.5%-103.3%, and the relative standard deviation within the day and between the day is less than 10%. The limit of detection (LOD) and the limit of quantitation (LOQ) in the feed are between 3.6-9.8 [mu] g kg <-1 > and 11.4-31.3 [mu] g kg <-1 >. The method is suitable for simultaneously determining three antibiotics, namely lincomycin, clindamycin and pirilimycin, in the feed. The method is accurate, sensitive, simple and convenient to operate and good in stability.
Description
technical field [0001] The invention relates to a method for measuring lincosamide antibiotics in feed. Background technique [0002] Lincosamide antibiotics are derived from the natural product lincomycin, including the semi-synthetic derivatives clindamycin and pirlinomycin ( figure 1 ) (Morar et al., 2009). In 1976, the first lincosamide antibiotic, lincomycin, was licensed for the first time in the United States for the treatment of infections caused by Gram-positive bacteria (Schwarz et al., 2016). Due to its narrow antibacterial spectrum, clindamycin was chemically modified to expand its antibacterial spectrum (Rezanka et al., 2007; Hoeksema et al., 1964), and clindamycin was approved by the Food and Drug Administration (FDA) in the US in 1970 approve. In 2000, a new lincosamide antibiotic, piromycin, was approved in the US and EU, which has been shown to be effective in the treatment of mastitis in lactating dairy cows (Jiang et al., 2015). It should be noted that...
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