Determination of Squalene in Banlan Leaves (Pulp, Juice, and Powder) by T/HIFSA 0006-2024

Scope

Scope: This standard specifies three methods for determining the content of squalene in Centella asiatica leaves (pulp, juice, and powder): gas chromatography, gas chromatography-mass spectrometry, and high-performance liquid chromatography. This standard applies to the determination of squalene in Centella asiatica leaves (pulp, juice, and powder), and other Centella asiatica products can be used as reference. When sampling 0.2g of the sample, the quantitative limit of gas chromatography is 30mg/kg, the quantitative limit of gas chromatography-mass spectrometry is 2.5mg/kg, and the quantitative limit of high-performance liquid chromatography is 30mg/kg. Main technical content: Method 1: Gas chromatography Principle: After saponification with potassium hydroxide-ethanol solution, the squalene in the sample is extracted with n-hexane and determined by gas chromatography with external standard quantification. Reagents and materials: Unless otherwise specified, all reagents are analytical grade, and the water is Class I water according to GB/T 6682. Reagents: - Potassium hydroxide (KOH). - Anhydrous ethanol (CH3CH2OH). - n-Hexane (C6H14): chromatograph-grade. - Preparation of potassium hydroxide-ethanol solution (about 2mol/L): Accurately weigh (112±0.1)g of potassium hydroxide, add 200mL of anhydrous ethanol, dissolve, and then dilute to 1000mL with anhydrous ethanol. - Standard squalene: Pure squalene standard (C30H50, CAS No.: 111-02-4), with a purity of ≥98.0%, or a standard substance certified and granted a standard material certificate by the state. - Preparation of standard solution: - Squalene standard stock solution (1.00mg/mL): Accurately weigh 100mg (accurate to 0.1mg) of squalene standard, dissolve in n-hexane, and transfer to a 100mL brown volumetric flask. Dilute to the mark and shake well to obtain a standard stock solution with a concentration of 1.00mg/mL. The standard stock solution can be stored at 4°C for 3 months in the dark

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