Method for separating and detecting impurity phenylhydrazine in edaravone sodium chloride injection
A technology for the detection of edaravone sodium chloride and its detection method, which is applied in the field of separation and detection of impurity phenylhydrazine in edaravone sodium chloride injection, can solve the problems of specificity that cannot meet the requirements and great interference, and achieve Good sensitivity, ensure the safety of medication, and the effect of simple preparation method
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Embodiment 1
[0035] The detection method of edaravone crude drug impurity phenylhydrazine in the Chinese Pharmacopoeia 2015 edition of embodiment 1;
[0036] Liquid chromatography conditions:
[0037] Instrument: Liquid Chromatograph-UV Detector;
[0038] Chromatographic column: Octadecylsilane bonded silica gel as filler;
[0039] Mobile phase: methanol-0.05mol / L ammonium dihydrogen phosphate solution (adjust the pH value to 3.5 with 20% phosphoric acid) (25:75);
[0040] Detection wavelength: 226nm.
[0041] Experimental steps:
[0042] Reference substance solution: take an appropriate amount of phenylhydrazine, accurately weighed, add mobile phase to dissolve and quantitatively dilute to make a solution containing about 0.15 μg per 1 ml, as the reference substance solution;
[0043] Need testing solution: get Edaravone Sodium Chloride Injection stock solution direct sample injection;
[0044] The test product spiked solution: take an appropriate amount of phenylhydrazine, accuratel...
Embodiment 2
[0046] Embodiment 2 phenylhydrazine detection wavelength screening
[0047] Using a diode array detector, scanning to obtain the ultraviolet absorption spectrum of phenylhydrazine see figure 2 ,Depend on figure 2 It can be seen that phenylhydrazine has maximum absorption wavelengths at 222nm and 274nm, and in the range of 222nm to 274nm, the absorption intensity gradually decreases with the increase of wavelength, and the mobile phase methanol is prone to terminal absorption at 220nm, which will interfere with the detection of impurity phenylhydrazine , Therefore, it is advisable to select a wavelength greater than 220nm as the detection wavelength, and the present invention selects 245nm as the detection wavelength.
Embodiment 3
[0048] Example 3 Elution Program Screening
[0049] In view of the fact that in Example 1, the unknown impurity peak interferes with the phenylhydrazine peak, and the specificity of the method is poor, try to optimize the chromatographic conditions.
[0050] Liquid chromatography conditions:
[0051] Experimental equipment: high performance liquid chromatography-ultraviolet detector;
[0052]Chromatographic column: InertSustain C18, 4.6mm×150mm, 5μm;
[0053] Mobile phase A: 0.05mol / L ammonium dihydrogen phosphate solution (adjust the pH value to 3.5 with 20% phosphoric acid);
[0054] Mobile phase B: Methanol;
[0055] Gradient elution procedure: use the following elution methods for investigation:
[0056] Elution mode-1
[0057]
[0058] Elution mode-2
[0059]
[0060] Elution mode-3
[0061]
[0062] Column temperature: 35°C;
[0063] Flow rate: 1.0ml / min;
[0064] Detection wavelength: 245nm;
[0065] Injection volume: 20μl;
[0066] Experimental step...
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