Determination of 8 kinds of macrolides and lincosamide residues in puffer fish and eels by liquid chromatography-tandem mass spectrometry

6.
2.
4.
1 Scope of application

Suitable fugu and eels lincomycin , oleandomycin , erythromycin , tilmicosin , tylosin , spiramycin , kitasamycin and cross residues Jos Liquid Chromatography - Tandem mass spectrometry
.

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2.
4.
2 Principle of the method

The residues of lincomycin, eleuomycin, erythromycin, tilmicosin, tylosin, spiramycin, kitasamycin and josamycin in puffer fish and eels are extracted with Tris buffer solution.
Purified by OasisHLB solid-phase extraction column, eluted with methanol, the eluent is concentrated to a constant volume, then determined by liquid chromatography-tandem mass spectrometry, and quantified by internal standard method
.

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2.
4.
3 Reagents and materials

Methanol, Acetonitrile : chromatographically pure; ammonium acetate , tris (hydroxymethyl) aminomethane (Tris, C _.
4 H _.
11 NO _.
3 ), calcium chloride (CaCI2 ₂ · 2H ₂ O), hydrochloric acid : analytically pure
.

Methanol solution: 2+3
.

The purity of the standard material is ≥95%
.

Standard stock solution I: 1.
0mg/mL
.

Standard working solution: 2.
0ug/mL
.

Internal standard stock solution: 1.
0mg/mL
.

Intermediate concentration internal standard solution: 10.
0μg/mL
.

Internal standard working solution: 1.
0μg/mL
.

Matrix standard mixed working solution for determination of puffer fish: draw 1.
0μL, 2.
0μL, 5.
0μL, 25.
0μL standard working solution respectively, add them to the corresponding reagent bottle in turn, and then add 20.
0μL internal standard working solution respectively, and use sample blank extract solution Make the volume to 1.
0mL
.
It is formulated into lincomycin, elipidomycin, erythromycin, tilmicosin, tylosin, spiramycin, kitasamycin and josamycin at 2.
0ng/mL, 4.
0ng/mL, A series of matrix standard mixed working solutions with four concentration levels of 10.
0ng/mL and 50.
0ng/mL;

Standard mixed working solution of matrix for determination of eel: draw 1.
0uL, 2.
0μL, 5.
0uL, 25.
0uL standard working solution of lincomycin, erythromycin, tylosin and kitatamycin and spiramycin and bamboo peach respectively 2.
5uL, 5.
0uL, 10.
0μL, and 25.
0μL standard working solutions of josamycin, josamycin, and tilmicosin, respectively, add them to the corresponding reagent bottle in turn, and then add 20.
0μL internal standard working solution respectively, and extract with sample blank The volume of the solution is set to 1.
0mL
.
It is formulated into lincomycin, erythromycin, tylosin, and kititamycin at 2.
0ng/mL, 4.
0ng/mL, 10.
0ng/mL, 50.
0ng/mL and spiramycin, elixiromycin, Josamycin and Tilmicosin are a series of matrix standard mixed working solutions with four concentration levels of 5.
0 ng/mL, 10.
0 ng/mL, 20.
0 ng/mL, and 50.
0 ng/mL, respectively
.

OasisHLB solid phase extraction column: 500mg, 6mL, or equivalent
.
Before use, it was activated with 10 mL methanol and 10 mL water successively, and kept the column moist in each step before vacuuming
.

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4.
4 Apparatus and equipment

Liquid chromatography-tandem mass spectrometer: equipped with electrospray ion source; balance: sensing volume 0.
01g, 0.
0001g; solid phase extraction device; nitrogen concentrator; polypropylene centrifuge tube with stopper: 50mL; centrifuge: 20000g centrifugal force (or Equivalent to 12000r/mi); ultrasonic cleaner; pH meter; oscillator
.

Related Links: Applicability of the method for determination of 8 macrolides and lincosamide residues in royal jelly and royal jelly freeze-dried powder