SN/T 4675.12-2016Determination of lysozyme in wine for export - HPLC method (English PDF)

Foreword

SN/T 4675 "export wine quality and safety analysis method" is divided into 30 parts.

--- SN/T 4675.1. Determination of glycerol in export wine enzymatic;

--- SN/T 4675.2. Determination of 2,3-butanediol in exported wine by gas chromatography;

--- SN/T 4675.3. Determination of ethanol stable carbon isotope ratio of exported wine;

--- SN/T 4675.4. Determination of lactic acid in export wine enzymatic;

--- SN/T 4675.5. Determination of organic acids in exported wine by ion chromatography;

--- SN/T 4675.6. Determination of glucose, fructose and sucrose in exported wine;

--- SN/T 4675.7. Determination of acetaldehyde in exported wine by gas chromatography - mass spectrometry;

--- SN/T 4675.8. Determination of 5-hydroxymethyl furfural in exported wine by liquid chromatography;

--- SN/T 4675.9. Determination of diethylene glycol in exported wine by gas chromatography - mass spectrometry;

--- Determination of ochratoxin A in export wines - Liquid chromatography - mass spectrometry/mass spectrometry;

--- SN/T 4675.11. Determination of 7 kinds of anthocyanins in exported wine by ultra performance liquid chromatography;

--- SN/T 4675.12. Determination of lysozyme in wine exports by liquid chromatography;

--- SN/T 4675.13. Determination of 2,4,6-trichloro-phenyl ether residues in exported wine by gas chromatography-mass spectrometry;

--- SN/T 4675.14. Determination of natamycin in wine exports by liquid chromatography - mass spectrometry/mass spectrometry;

--- SN/T 4675.15. Determination of salicylic acid, dehydroacetic acid and p-chlorobenzoic acid in exported wine by liquid chromatography;

--- SN/T 4675.16. Determination of fumaric acid in exported wine by liquid chromatography - mass spectrometry/mass spectrometry;

--- SN/T 4675.17. Determination of butyl tin content in exported wine - Gas Chromatography - Mass Spectrometry/Mass Spectrometry;

--- SN/T 4675.18; Determination of dithiocarbamate residues in exported wine by headspace gas chromatography;

Determination of sodium, magnesium, potassium, calcium, chromium, manganese, iron, copper, zinc, arsenic, selenium, cadmium, lead in exported wine;

--- SN/T 4675.20. Determination of rare earth elements in exported wine by inductively coupled plasma mass spectrometry;

--- SN/T 4675.21. Determination of soluble inorganic salts in export wine by ion chromatography;

--- SN/T 4675.22. Determination of total sulfur dioxide in wine exports by colorimetry;

--- SN/T 4675.23. Determination of ammonia nitrogen in export wines and grape juice - Continuous flow analyzer method;

--- SN/T 4675.24. Determination of export wine Fulin - schoca index spectrophotometer;

--- SN/T 4675.25. Determination of the color of exported wine CIE1976 (L * a * b *) color space method;

--- SN/T 4675.26. Determination of turbidity of export wine by scattered light method;

--- SN/T 4675.27. Determination of export wine alkaline ash;

--- SN/T 4675.28. Exports of wine bacteria, mold and yeast count;

--- SN/T 4675.29. Export wine wine yeast test real-time fluorescent PCR;

--- SN/T 4675.30. Exported wine Bayer engaged yeast test real-time PCR method.

This section is part 12 of SN/T 4675.

This section drafted in accordance with GB/T 1.1-2009 given rules.

This part of the revision uses the International Grape and Wine Organization (OIV) OIV-MA-AS315-14 "Determination of lysozyme in wine - liquid Phase chromatography. " This section is technically compatible with this method, only the individual content of the OIV method OIV-MA-AS315-14 the editorial changes, the main content of the amendment.

--- Optimization of chromatographic conditions.

Please note that some of this document may be patentable. The issuing agencies of this document do not bear the responsibility of identifying these patents.

This part of the national certification and accreditation supervision and management committee proposed and centralized.

This part of the drafting unit. People's Republic of China Guangdong Exit Inspection and Quarantine, People's Republic of China Beijing Exit Inspection and Quarantine Epidemic

1 Scope

This part of SN/T 4675 specifies the method for the determination of lysozyme content in wine by high performance liquid chromatography.

This section applies to white wine and sparkling wine Determination of lysozyme content.

2 Normative references

The following documents for the application of this document is essential. For dated references, only the dated version applies to this article Pieces. For undated references, the latest edition (including all amendments) applies to this document.

GB/T 6682 analytical laboratory water specifications and test methods

3 Method summary

This method only detects the content of lysozyme with enzymatic activity in wine, but has nothing to do with the true activity of the enzyme in the protein.

Lysozyme in wine after reversed-phase phenyl column separation, high performance liquid chromatography with fluorescence detector or UV detector, retention time Qualitative, external standard method.

4 Reagents and materials

Unless otherwise specified, the reagents used are of analytical grade, water GB/T 6682 provisions of a water.

4.1 Acetonitrile (C2H3N). Chromatographic purity.

4.2 anhydrous ethanol (C2H6O).

Hydrochloric acid (HCL).

4.4 tartaric acid (C4H6O6).

4.5 Trifluoroacetic acid (TFA).

4.6 5% hydrochloric acid solution. Take 5mL hydrochloric acid, water to volume 100mL.

4.7 tartaric acid solution Weigh 1.0g tartaric acid (4.4), with 100mL of anhydrous ethanol was dissolved, the volume of water to 1000mL.

4.8 Lysozyme standard material (CAS. 12650-88-3).

4.9 lysozyme standard stock solution accurately weighed amount of lysozyme standard (4.8), add appropriate tartaric acid solution (4.7) ultrasonic oscillation to charge Divide and dilute to volume, prepare a standard stock solution with a concentration of 2µL and store in a refrigerator at 0 °C ~ 4 °C.

4.10 Lysozyme standard working solution, respectively, the exact amount of lysozyme standard stock solution (4.9), diluted with the sample extract to prepare 5.0mg/L, 10.0mg/L, 20.0mg/L, 25.0mg/L, 40.0mg/L, 50.0mg/L, 60.0mg/L standard working solution, is with current use.

4.11 Microporous membrane .0.22µm, water system.

5 Instruments and equipment

5.1 High performance liquid chromatography with fluorescence detector or UV detector.