T/NXSLX 0907-2024 Determination of hydroxy saffron yellow pigment A content in the stems and leaves of red saffron

Scope

Scope: This document specifies the terms, definitions, principles, reagents, materials, instruments, equipment, sample collection and preparation, determination steps, and result calculation for the determination of hydroxy saffron yellow pigment A in saffron stems and leaves in Ningxia region. This document is applicable to the determination of hydroxy saffron yellow pigment A in saffron stems and leaves in Ningxia region. The detection limit is 12.5 μg/mL to 800 μg/mL. Main technical content: High-performance liquid chromatography (HPLC) is based on classical liquid chromatography. A liquid is used as the mobile phase, and different polar solvents are pumped into a high-efficiency stationary phase chromatographic column with extremely fine particles. The components are separated and detected by the detector, enabling rapid and highly sensitive detection of samples. Hydroxy saffron yellow pigment A in saffron stems and leaves is detected and verified by this method. 7. Determination steps 7.1 Sample preparation: Take saffron stems and leaves samples and grind them in a multifunctional crusher. Sieve through a 20-mesh sieve for later use. 7.2 Preparation of liquid solutions 7.2.1 Standard solution preparation: Precisely weigh 8 mg of hydroxy saffron yellow pigment A standard sample and dissolve it in 25% methanol to a volume of 10 mL in a volumetric flask, obtaining an 800 μg/mL hydroxy saffron yellow pigment A standard control solution. 7.2.2 Sample solution preparation: Precisely weigh 0.5 g of sieved saffron stem and leaf powder, add 125 mL of 57% methanol, and extract for 45 minutes at 70°C and 80 Hz (250 W) ultrasound. Filter the solution through a 0.22 um filter membrane to obtain the saffron stem and leaf sample solution. 7.3 Sample determination: Inject 20μL of the sample solution under the following conditions: mobile phase: 0.2% phosphoric acid water (A) - acetonitrile (B) - methanol (D), gradient elution: 0–6.5 min, 80% A - 9% B - 11% D; 6

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