Method for separating and determining calcipotriene starting raw material A and related impurities

[0072] 1. Instruments and conditions

[0073] Instrument: high performance liquid chromatography;

[0074] Chromatographic column: Agilent ZORBAX SIL (4.6×250mm, 5μm);

[0075] Mobile phase: n-hexane:isopropanol=90:10(V:V);

[0076] UV detector detection wavelength: 200nm;

[0077] Column temperature: 30°C;

[0078] Flow rate: 1.0ml / min;

[0079] Injection volume: 10μl;

[0080] Diluent: The mobile phase is n-hexane:isopropanol=90:10 (V:V).

[0081] 2. Experimental steps

[0082] (1)A 1 Reference substance solution stock solution: Accurately weigh A 1 100.01mg plus diluent dissolved to make 5.05mg / mL A 1 Reference substance solution stock solution;

[0083] (2)A 2 Reference substance solution stock solution: Accurately weigh A 2 102.55mg plus diluent dissolved to make 5.12mg / mL A 2 Reference substance solution stock solution;

[0084] (3)A 3 Reference substance solution stock solution: Accurately weigh A 3 101.27mg plus diluent dissolved to make 5.06mg /

[0092] 1. Instruments and conditions

[0093] Instrument: high performance liquid chromatography;

[0094] Chromatographic column: Agilent ZORBAX SIL (4.6×250mm, 5μm);

[0095] Mobile phase: n-hexane:isopropanol=95:5(V:V);

[0096] Evaporative photodetector conditions: drift tube temperature: 51.0°C;

[0097] Gas flow rate: 1.7L / min;

[0098] Air pressure: 0.5Mpa;

[0099] Column temperature: 30°C;

[0100] Flow rate: 1.0ml / min;

[0101] Injection volume: 10μl;

[0102] Diluent: The mobile phase is n-hexane:isopropanol=95:5 (V:V).

[0103] 2. Experimental steps

[0104] (1)A 1 Reference substance solution stock solution: Accurately weigh A 1 100.01mg plus diluent dissolved to make 5.05mg / mL A 1 Reference substance solution stock solution;

[0105] (2)A 1 Positioning solution: precise measurement of A 1 Put 1.0ml of the reference substance solution stock solution in a 10ml measuring bottle, add diluent to dilute to constant volume, shake well, and obtain;

[01

[0120] 1. Instruments and conditions

[0121] Instrument: high performance liquid chromatography;

[0122] Chromatographic column: Agilent ZORBAX SIL (4.6×250mm, 5μm);

[0123] Mobile phase: n-hexane:isopropanol=99.5:0.5(V:V);

[0124] Evaporative photodetector conditions: drift tube temperature: 42°C;

[0125] Gas flow rate: 1.7L / min;

[0126] Air pressure: 0.5Mpa;

[0127] Column temperature: 30°C;

[0128] Flow rate: 1.0ml / min;

[0129] Injection volume: 10μl;

[0130] Diluent: the mobile phase is n-hexane:isopropanol=99.5:0.5 (V:V).

[0131] 2. Experimental steps

[0132] (1)A 5 Reference substance solution stock solution: Accurately weigh A 5 100.18mg plus diluent dissolved to make 0.501mg / mLA 5 Reference substance solution stock solution;

[0133] (2)A 5 Positioning solution:: Precise measurement of A 5 Put 1.0ml of the stock solution of the reference substance solution in a 10 volumetric bottle, add diluent to the mark, shake well, and obtain;

[0134