High performance liquid chromatography analysis method of urea and its impurity carbamylurea, methylene biuret
A technology of high performance liquid chromatography and methylene diurea, which is applied in the field of high performance liquid chromatography analysis method, can solve the problems of time-consuming, high operation cost and high requirements for inspection personnel, and achieves low detection limit, high sensitivity and precision. Good results
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Embodiment 1
[0023] laboratory apparatus:
[0024] LC-10ATVP high performance liquid chromatography (Shimadzu, Japan);
[0025] SPD-10ATVP UV-Vis detector (Shimadzu, Japan);
[0026] Hangzhou Puhui Technology Co., Ltd. Sepu 3000P chromatography workstation;
[0027] Ultrasonic degasser (Shanghai Ultrasonic Co., Ltd.);
[0028] Filter: filter membrane pore size 0.45μm.
[0029] Chromatographic conditions:
[0030] Chromatographic column: 250mm×4.6mm(id) stainless steel column, built with Kromasil-5μm-C 18 filler;
[0031] The mobile phase composition volume ratio: methanol: water = 3: 97, wherein the water is desalted water, and boiled, methanol is analytically pure, the mobile phase is filtered with a 0.45 μm filter membrane and degassed with ultrasonic waves for 30 minutes before use;
[0032] Flow rate: 0.35Ml / min;
[0033] Column temperature: room temperature;
[0034] Detection wavelength: 210nm;
[0035] Injection volume: 6μl;
[0036] Internal standard: melamine.
[0037] ...
Embodiment 2
[0090] Chromatographic conditions:
[0091] Mobile phase composition volume ratio: methanol: water = 10:90;
[0092] Flow rate: 0.30Ml / min;
[0093] Column temperature: room temperature;
[0094] Detection wavelength: 215nm;
[0095] Injection volume: 5μl;
[0096] Internal standard: melamine.
[0097] All the other implementation modes are the same as the first embodiment.
[0098] (1). Methylene diurea linear relationship:
[0099] Using samples containing different concentrations of methylene diurea, the detection curve equation of methylene diurea is C / C i =-0.107551+4.34741×A / A i .
[0100] (2). Urea linear relationship
[0101] Prepare urea samples with different concentrations, and the curve equation of urea obtained by detection is C / C i =-16.0711+190.29×A / A i .
[0102] (3). Biuret linear relationship
[0103] Select biuret samples with different concentrations, and the curve equation of urea is C / C i =-0.00752131+7.53833×A / A i .
[0104] Pipette 6 μl ...
Embodiment 3
[0126] Chromatographic conditions:
[0127] Mobile phase composition volume ratio: methanol: water = 4: 96;
[0128] Flow rate: 0.38Ml / min;
[0129] Column temperature: room temperature;
[0130] Detection wavelength: 205nm;
[0131] Injection volume: 7μl;
[0132] Internal standard: melamine.
[0133] All the other implementation modes are the same as the first embodiment.
[0134] (1). Methylene diurea linear relationship:
[0135] Using samples containing different concentrations of methylene diurea, the detection curve equation of methylene diurea is C / C i =-0.107541+4.34744×A / A i 4.
[0136] (2). Urea linear relationship
[0137] Prepare urea samples with different concentrations, and the curve equation of urea obtained by detection is C / C i =-16.0711+190.32×A / A i .
[0138] (3). Biuret linear relationship
[0139] Select biuret samples with different concentrations, and the curve equation of urea is C / C i =-0.00752128+7.53817×A / A i .
[0140] Pipette 6 μl...
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