T/QAS 013-2020 Determination of rutin content in Goji berries by high performance liquid chromatography (HPLC) method

Scope

Scope: This standard specifies the principles, reagents, instruments and equipment, sample preparation, analysis steps, data processing, and precision for the determination of rutin in wolfberry by high-performance liquid chromatography (HPLC). This standard applies to the determination of rutin in dried wolfberry. Main technical content: 6. Sample preparation: - Take a representative sample of 200 g and reduce it to 100 g by the four-division method. - Before testing, take 100 g of the sample and dry it in a 60°C oven for more than 5 hours. Then remove the sample, cool it to room temperature, and grind it using a high-speed universal crusher. Place it in a sample bag for later use. 7. Analysis steps: 7.1 Sample extraction: - Accurately weigh 1.0 g (with an accuracy of 0.0001 g) of ground wolfberry into a 100 mL centrifuge tube. Add 50 mL of 70% (v/v methanol/water) methanol solution, shake well, and place it in an ultrasonic cleaner for 50 min of ultrasonic extraction (shaking once every 15 min). After removal, centrifuge at 4000 rpm for 10 min. Filter the supernatant through a 0.45 μm microporous filter membrane and prepare it for HPLC analysis. 7.2 HPLC reference conditions: - Column: Hypersil GOLD C18 column (250 mm × 4.6 mm, 5.0 μm), or equivalent; - Column temperature: 30°C; - Mobile phase: A: methanol, B: 4% formic acid solution, A:B=40%:60%; - Flow rate: 1.0 mL/min; - Wavelength: 360 nm; - Injection volume: 10 μL. 7.3 Plotting the standard curve: - Dilute the rutin standard solution with 70% methanol solution to obtain standard working solutions with concentrations of 0, 5, 10, 50, 100, and 200 μg/mL (see Table 1). Inject and detect the samples in order from low to high, and plot the mass concentration X (μg/mL) on the horizontal axis and the response value (peak area) Y on the vertical axis to perform linear regression and obtain the standard curve regression equation. Table 1: S

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