GB 23200.101-2016Food safety national standard -- Determination of residues of acaricides in royal jelly by gas chromatography -- Mass spectrometry (English PDF)

4.1 Reagents

4.1.1 ether (C4H10O). pure chromatography.

4.1.2 n-hexane (C6H14). pure chromatography.

4.1.3 Acetone (C3H6O). Chromatographic pure.

4.1.4 Sodium chloride (NaCl).

4.1.5 anhydrous sodium sulfate (Na2SO4). 650 °C burning 4h, in the dryer to cool to room temperature, stored in a sealed bottle in reserve.

4.2 Standards

4.2.1 acaricidal mite, methyl acaricides,, deer mites, ethyl methicillin, bromocarbide, dicofol, Purity >=99%, see Appendix A.

4.3 Standard solution preparation

4.3.1 acaricides, methyl methicillin, acaricides, lemi mite, ethyl methicillin, bromide mite, dicofol, Solution. Weigh the appropriate amount of standard material accurately, with n-hexane dubbed the concentration of 100 µl/mL standard stock solution. Store in 4 °C refrigerator.

4.3.2 acaricides, methicillin mites, acaricides, lei mites, ethyl methicillin, bromocarbide, dicofolidone, pyridaben standard work Liquid. Dilute the mixture with n-hexane to the appropriate concentration of the standard working solution. Store in 4 °C refrigerator.

4.4 Materials

4.4.1 Florisil Solid Phase Extraction Column. (1 g, 6 mL) or equivalent. Use a 10 mm high anhydrous sodium sulphate layer in the front column with 5 mL N - hexane elution activated solid phase extraction.

7.1 Extraction

Weigh 2 g sample (accurate to 0.01 g) in 50 mL centrifuge tube, add 10 mL water vortex mixed 1 min, standing 5 min. Join 20 mL of n-hexane acetone (1 1, volume ratio) mixed solvent, vortex mixed for 1 min, centrifuged at 4000 r/min for 3 min, the upper organic Phase transferred into the concentration bottle. Add 10 mL of n-hexane acetone (1 1, volume ratio) mixed solvent, repeat the extraction time, combined the upper organic phase, in 45 °C below the water bath under reduced pressure to near dry, to be purified.

7.2 Purification

The residue in the concentrated flask was washed twice with 3 mL of n-hexane and transferred to a Florisil solid phase extraction column. With 5 mL of n-hexane the effluent was discarded and eluted with 10 mL of n-hexane + ether (8 + 2, v/v) mixed solvent. The eluate was collected in 10 mL of glass off Tube in the 45 °C water bath with nitrogen blowing instrument slowly blowing to near dry, with n-hexane dissolved and set to 0.5mL, gas chromatography-mass spectrometry.

7.3 Determination

7.3.1 Gas Chromatography - Mass Spectrometry Reference Conditions

A) Column. DB-5ms quartz capillary column, 30 m x 0.25 mm (id) x 0.25 µm, or equivalent;

B) column temperature. 50 °C for 1 min, at 20 °C/min heating rate rose to 220 °C, keep 1 min, the temperature was raised to 250 ° C at a rate of 5 ° C/min and raised to 280 ° C at a rate of 20 ° C/min for 8 min.

C) Inlet temperature. 250 ° C;

D) chromatographic - mass spectrometer interface temperature. 280 ° C;

E) Carrier gas. helium, purity greater than or equal to 99.999%; flow rate 1.0 mL/min;

F) Injection volume. 2 µL;

G) Injection method. splitless injection, 1.5 min after the opening valve;

H) ionization mode. NCI;

I) ionization energy. 184 eV;

J) ion source temperature. 150 ° C;

K) quadrupole temperature. 150 ° C;

L) Reaction gas. methane, purity greater than or equal to 99.99%, reaction gas flow rate. 2 mL/min;

M) Measurement method. Select the ion monitoring mode (SIM);

N) solvent delay time. 10 min;

O) Select the ion measurement. each compound selected a quantitative ion, 2-3 qualitative ions. Each group needs to be measured Sub-according to the peak order, respectively, time points were measured. The retention time of each compound, the quantitative ion, the qualitative ion and the quantitative ion and qualitative the abundance of ions is given in Appendix B.

7.3.2 Quantitative determination

According to the content of acaricide in the sample solution, the standard working solution with similar peak area is selected. Standard working solution and sample solution of acaricides the value should be within the linear range of the instrument determination. Standard working solution and sample volume and other volume interspersed injection determination. Under the above chromatographic conditions, 8 species the retention time of the acaricides is given in Appendix B, and the total ion current of the standard is shown in Appendix C, C.1.

7.3.3 Qualitative determination

The standard solution and sample solution are determined according to the conditions specified in 7.3.1, if the sample solution and the standard solution in the same retention time peak Now, it is confirmed by mass spectrometry, after deducting the background of the sample spectrum, the selected ions all appear, while the selected ion from the the abundance ratio is consistent with the relative abundance of the ions associated with the standard, and the fluctuation range is within the maximum allowable deviation of Table 1 (see Table 1) the substance is present in the sample.

Table 1 Mass Spectrum Relative Ion Abundance Maximum Allowable Deviation

Relative ion abundance (% base) > 50% > 20% to 50% > 10% to 20% <=10% Allowable relative deviation ± 20% ± 25% ± 30% ± 50%

7.4 Blank test

In addition to the sample, according to the above determination steps.

10.1 Quantitation limits

The quantification limit of this method is. 0.01 mg/kg.

10.2 Recovery rate

The experimental data on the concentration and recovery of the sample are given in Appendix D.

Appendix A

(Informative)

8 kinds of acaricides pesticides in English generic name, chemical formula, CAS number information

Table A.1 8 kinds of acaricides pesticides in English common name, chemical formula, CAS No. information

Chinese generic name English generic name chemical molecular formula CAS number Acetylidene Chlorbenside C13H10 Cl2 S 269 103-17-3 Mycobacterium acaricides Quinomethionate C10H6N2OS2 234 2439-01-2 Acetylate Chlorfenson C12H8Cl2O3 S 302 80-33-1 Phytophthora Ethyl Acetone Chlorobenzilate C16H14Cl2O3 324 510-15-6 Bromopropylate C17H16Br2O3 426 18181-80-1 Dicofolone Tetradifon C12H6 Cl4 O2S 354 116-29-0 Pyridaben C19H25ClN2OS 364 96489-71-3 Appendix B (Informative appendix) Retention time of 8 kinds of acaricides, quantitative ions, qualitative ions and relative abundance

Table B.1 Retention times, quantitative ions, qualitative ions and relative abundance of eight acaricides

Pesticide name quantitative ion qualitative ion 1 qualitative ion 2 qualitative ion 3 keep time (Min) Acaricides 143 (100) 144 (7.7) 145 (36.1) 13.5 Mycobacterium mites 206 (100) 207 (12.8 208 (11.8) 13.6 Acaricides 175 (100) 176 (8.3) 177 (38.9) 14.0 Lychee mite 223 (100) 224 (16.1) 239 (46.6) 14.6 Ethyl Ester 324 (100) 326 (68.2) 278 (34.5) 262 (40.2) 15.1 Bromide mites 428 (100) 426 (51.) 430 (50.4) 17.5 Diclofenac 320 (100) 318 (75.4) 356 (43.9) 18.1 Pyridaben 217 (100) 219 (39.1) 218 (15.8) 19.8 Appendix C (Informative) Total ion flow chart of 8 kinds of acaricides standard materials

Figure C.1 Total ion chromatogram of eight species of acaricides

1. Acaricidal ether 2. Methylk acaricides 3. Fungicides 4. Lecture dex mite 5 Ethyl ester Mites 4. Bromothylate 7. Diclofenac 8 Appendix D (Informative) Sample concentration and recovery of the experimental data

Table D.1 Experimental data on the concentration and recovery of the sample

Pesticide name added concentration (Mg/kg) Recovery rate Precision degree Pesticide name Add concentration (mg/kg) Recovery rate Precision degree Acaricides 0.01 86.7 ~ 112 11.2 ethyl ester acaricides 0.01 77.5 to 113 6.8 0.02 94 to 112 8.6 0.02 84.0 to 104 7.9 0.05 81.2 to 107 9.3 0.05 72.6 to 107 4.5 Methyl killer 0.01 82.1 ~ 118 9.6 Bromo mite 0.01 71.3 ~ 115 13.0 0.02 76.8 to 107 10.2 0.02 78.0 to 114 11.8 0.05 79.4 to 102 7.5 0.05 73.6 to 106 10.8 Acaricidal ester 0.01 79.7 ~ 118 13.1 dicofol 0.01 76.7 to 115 6.9 0.02 85.0 to 110 12.6 0.02 75.0 to 107 7.6 0.05 74.4 to 109 10.8 0.05 74.8 to 108 8.9 Lymanthamidae 0.01 69.1 to 108 14.1 Pyridinidone 0.01 78.2 to 118 8.4 0.02 76.5 to 108 10.6 0.02 75.0 to 109 9.8 0.05 77.4 to 109 13.1 0.05 78.8 to 109 12.3 Appendix E (Normative appendix) Laboratory repeatability requirements

Table E.1 Laboratory repeatability requirements

Measured component content Mg/kg Precision <=0.001 36 > 0.01 > 1 14 Appendix F (Normative appendix) Inter-laboratory reproducibility requirements

Table F.1 Inter-laboratory reproducibility requirements

Measured component content Mg/kg Precision <=0.001 54 > 0.01 > 1 19