SN/T 5149-2019Determination of cabergoline residues in foodstuffs of animal origin for export LC-MS/MS method (English PDF)

Foreword

This standard was drafted in accordance with the rules given in GB/T 1.1-2009.

Please note that some content of this document may involve patents, the publication of this document the agency is not responsible for identifying these patents.

This standard is proposed and managed by the General Administration of Customs of the People's Republic of China.

This standard was drafted. Shijiazhuang Customs, People's Republic of China.

1 Scope

This standard specifies the liquid chromatography-mass spectrometry/mass spectrometry method for cabergoline in foods of animal origin for export.

This standard applies to the detection of cabergoline residues in animal source foods such as chicken, pork, beef, beef fat, beef liver and milk.

2 Normative references

The following documents are essential for the application of this document. For dated references, only the dated version applies to this document.

file. For undated references, the latest version (including all amendments) applies to this document.

GB/T 6682 Analytical laboratory water specifications and test methods

3 Principle

Cabergoline in the sample was extracted with acetonitrile, degreased with acetonitrile-saturated n-hexane, purified on a neutral alumina column, and liquid chromatography-mass spectrometry /Determined by mass spectrometer and quantified by internal standard method.

4 Reagents and materials

Unless otherwise specified, all reagents are of analytical grade, and water is the first-grade water specified in GB/T 6682.

4.1 Acetonitrile. HPLC grade.

4.2 n-hexane. HPLC grade.

4.3 Methanol. HPLC grade.

4.4 Formic acid. HPLC grade.

4.5 Anhydrous sodium sulfate.

4.6 0.1% formic acid. exactly 1.0mL of formic acid (4.4) was dissolved in 1L of water.

4.7 0.1% formic acid water-acetonitrile solution (9 + 1, v/v). Measure 90 mL of 0.1% formic acid aqueous solution (4.6) and 10 mL of acetonitrile, and mix.

4.8 Acetonitrile-saturated n-hexane solution. 100 mL of n-hexane was added to 20 mL of acetonitrile, and after shaking sufficiently, the layers were left to stand and the upper layer of n-hexane was taken.

The alkane layer is reserved.

4.9 Cabergoline (CAS. 81409-90-7, molecular formula. C26H37N5O2). purity is greater than 99.0%.

4.10 Cabergoline-D5. Purity greater than 99.0%.

4.11 Standard stock solution (100 µg/mL). Weigh accurately 10 mg of cabergoline standard (accurate to 0.1 mg), dissolve with methanol and make up to 100 mL, and prepare a standard stock solution of 100 µg/mL. This solution is stored at -18 ° C, protected from light, and has a validity of 3 months. 4.12 Standard intermediate solution (1µg/mL). Accurately transfer the standard stock solution from 1.0mL (4.11) to a 100mL volumetric flask, make up to volume with methanol, and prepare a concentration of 1µg/mL, below -18 °C Keep away from light, valid for one month. 4.13 Standard working solution. Dilute the standard intermediate solution to the required standard working solution step by step with a constant volume solution as required. The standard working solution requires Active use.

4.14 Standard internal standard stock solution (10 µg/mL). Accurately weigh 1 mg of cabergoline-D5 standard (accurate to 0.1 mg) and dissolve it in methanol.

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Dissolve and dilute to 100 mL, and prepare a standard internal standard stock solution of 10 µg/mL. This solution is stored at -18 ° C, protected from light, and has a validity of 3 months. 4.15 Standard internal standard working solution (100ng/mL). Accurately transfer the standard internal standard intermediate solution from 0.5mL (4.14) to a 50mL volumetric flask.

Make up the volume of methanol to the mark, prepare a concentration of 100ng/mL, and store at -18 ° C in the dark, and the solution is valid for one week.

4.16 Solid-phase extraction column. Neutral alumina column (Alumina-N), 1000 mg, 3 mL.

4.17 Microporous filter membrane. 0.22 µm, organic phase needle filter.

5 Instruments and equipment

5.1 Liquid chromatography-mass spectrometry/mass spectrometer. Power distribution spray ion source (ESI).

5.2 Analytical balance. Sensitivity is 0.01g and 0.1mg, respectively.

5.3 Homogenizer, rotation speed is not less than 1500r/min.

5.4 Centrifuge, the rotation speed is not less than 4,000r/min.

5.5 Vortex mixer.

5.6 Tissue masher.

5.7 Solid-phase extraction device.

5.8 Nitrogen blowing instrument.

6 Sample preparation and storage

7 Analysis steps

8 Calculation and presentation of results

Using the internal standard method in the data processing software, calculate the residual amount of cabergoline in the sample according to formula (1).

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In the formula.

X --- the content of cabergoline in the sample, the unit is micrograms per kilogram (µg/kg); R --- the ratio of the peak area of the analyte in the sample to the internal standard;

--- the concentration of the analyte in the standard solution, the unit is nanogram per milliliter (ng/mL);

--- the final constant volume of the sample solution, in milliliters (mL);

RS --- The ratio of the peak area of the analyte in the standard solution to the internal standard;

Note. The calculation result should be deducted from the blank value.

9 Limit of quantitation and recovery

9.1 Limit of quantification.

The limit of quantification of cabergoline by this method is 0.1 µg/kg.

9.2 Recovery rate range.

Partial recovery experimental data are shown in Table 3.

Table 3 Adding recovery rate of cabergoline (state = 8)

Food name added concentration (µg/kg) average value (Mg/kg) Recovery rate (%) beef 0.10.095 84.0 to 112.0 0.2 0.187 85.5 to 105.5 1.0 0.970 88.0 to 105.0 pork 0.10.096 87.0 to 105.0 0.2 0. 18.8 88.0 to 101.0 1.0 0.990 89.8 to 115.0 chicken 0.1 0.093 81.0 to 102.0 0.2 0.186 84.5 to 101.5 1.0 0.980 87.0 to 111.0 Cow fat 0.1 0.093 89.0 to 98.0 0.2 0.292 88.0 to 105.0 1.0 0.970 83.0 to 108.0 Bovine liver 0.10.095 88.0 to 105.0 0.2 0. 18.8 84.5 to 104.0 1.0 0.960 83.0 to 112.0 milk 0.10.095 84.0 to 103.0 0.2 0. 18.8 81.0 to 102.5 1.0 0.960 85.0 to 110.0 SN/T 519-2019 Appendix A (Informative appendix) MS Reference Conditions

a) Electrospray voltage (kV). 0.5;

b) Taper hole voltage (V). 20;

c) Desolvent gas temperature (° C). 500;

d) Solvent gas flow rate (L/Hr). 1000;

e) Conical hole back-blow flow rate (L/Hr). 150

f) Nebulizer pressure (kPa). 700;

g) collision gas. high purity argon;

h) Velocity of collision gas (mL/min). 1.7;

i) For other mass spectrometry parameters, see Table A. 1.

Table A. 1 Multiple reaction monitoring conditions

Analyte retention time (min) Ion pair (M/z) Collision energy (CE)/ev Cabergoline 2.54 452.5/381.4 17 452.5/279.3 3 24 Cabergoline-D5 2.53 457.5/386.5 18 . Quantitative ions (theionforquation).

Non-commercial statement. The reference mass spectrometry conditions listed in Appendix A were completed on a Watters TQS LC/MS instrument. The test instrument types listed here are only it is provided for reference and does not involve commercial purposes. Standard users are encouraged to try different manufacturers or models of instruments.

SN/T 519-2019

Appendix B

(Informative appendix) Standard chromatogram Cabergoline Cabergoline-D5

Figure B. 1 LC-MS/MS chromatogram of cabergoline standard

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0.00 90 10

1.90 90 10

5.90 10 90

6.00 90 10

7.90 90 10

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In the formula.

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0.10.095 84.0 to 112.0 0.2 0.187 85.5 to 105.5 1.0 0.97 88.0 to 105.0 0.10.096 87.0 to 105.0 0.2 0. 18.8 88.0 to 101.0 1.0 0.999 89.8 to 115.0 0.1 0.093 81.0 to 102.0 0.2 0.186 84.5 to 101.5 1.0 0.98 87.0 to 111.0 0.1 0.093 89.0 to 98.0 0.2 0.292 88.0 to 105.0 1.0 0.97 83.0 to 108.0 0.10.095 88.0 to 105.0 0.2 0. 18.8 84.5 to 104.0 1.0 0.96 83.0 to 112.0 0.10.095 84.0 to 103.0 0.2 0. 18.8 81.0 to 102.5 1.0 0.96 85.0 to 110.0 SN/T 519-2019 17 452.5/279.3 3 24 SN/T 519-2019 cabergoline caberline-D5 SN/T 519-2019