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This standard specifies the method for determining the content of chondroitin sulfate and nitroglucose hydrochloride using the liquid phase colorimetric method. This standard is applicable to the determination of chondroitin sulfate and glucosamine hydrochloride extracted from animal cartilage tissues such as cows, pigs, and sharks. 2 Normative References: The clauses in the following standard documents become clauses of this standard through reference in this standard. For dated reference documents, all subsequent amendments (excluding corrected content) or revisions shall not apply to this standard. However, all parties who have reached an agreement based on this standard shall study whether the latest versions of these documents can be used. For undated reference documents, the latest versions shall apply to this standard. GB/T 6682 Specification and Test Methods for Analytical Laboratory Water (GB/T 6682-1992, neq 1SO 36961987)
3 Principle: Use acetonitrile to disperse the chondroitin sulfate sample evenly, dissolve it in water, separate it by liquid chromatography, and measure it at a wavelength of 192 nm using external standard method. 4 Reagents and Materials 4.1 Water: Grade 1 water according to GB/T 6682. 4.2 Acetonitrile; Liquid chromatography grade. 4.3 Sodium pentane sulfonate; Purity ≥ 99.5%. 4.4: chondroitin sulfate A standard:; The purity is about 70%, and the rest is chondroitin sulfate C. The structural formula is shown in Appendix A. The purity of standard substance Z of 4.5 sulfate cartilage Jing B is about 90%, and the rest are sulfate cartilage flocs A and C. The structural formula is shown in Appendix A. 4.6 Standard substance of chondroitin sulfate C: The purity is about 90%, and the rest is chondroitin sulfate floc A. The structural formula is shown in Appendix A.4.7 Glucosamine hydrochloride; Purity ≥ 99.0%. 4.8 Standard solution: Weigh 25.0 mg of chondroitin sulfate standard A (or B, or C) and 25.0 mg of glucosamine hydrochloride dried at 105 ℃ for 4 hours into a beaker, add 5.0 mL of acetonitrile, shake to disperse evenly, then add water, dissolve with ultrasound, transfer to a 50 mL volumetric flask, and bring to volume with water. The concentration of chondroitin sulfate is about 0.5 mg/mL, and the concentration of glucosamine hydrochloride is about 0.5 mg/mL 4.9 Sodium pentane sulfonate solution; Weigh 0.87 g of sodium pentane sulfonate solution and dissolve it in 500 mL of water. The concentration of the solution is 10 mmol/L, and use a 5.1 ultrasonic generator and equipment. 5.2 Analytical balance: with a sensitivity of 0.1 ng. 5.3 High performance liquid chromatography: equipped with a diode array detector or ultraviolet detector. 5.4 Filter membrane: PTFE membrane, 0.2 um
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