Method for measuring content of arginine in rhTNK-tPA (recombinant human TNK tissue-type plasminogen activator) with HPLC (high performance liquid chromatography)
A high-performance liquid chromatography and arginine technology, which is applied in the field of high-performance liquid chromatography to determine the content of arginine in Mingfule, can solve the problems of short peak time, high requirements for detection equipment, and cumbersome operation, and achieve quality improvement Effect
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Embodiment 1
[0048] Embodiment 1, the assay method of arginine content in Mingfule
[0049] Chromatographic conditions:
[0050] Chromatographic column: C8 acrylamide bonded silica gel column;
[0051] Mobile phase: 0.05mol / L potassium dihydrogen phosphate solution: acetonitrile=40:60, pH is 7.0, in parts by volume;
[0052] Detection wavelength: 208mm;
[0053] Column temperature: 30°C;
[0054] Flow rate: 1.0ml / min;
[0055] Injection volume: 10 μl.
[0056] Determination steps: take 1 bottle of the test sample and reconstitute it according to the marked amount, dilute it 10 times with the mobile phase and add the standard product (about 1.375g) into the mobile phase to dissolve, so as to obtain the test sample solution and the standard product solution, take 10 μl each, and inject them respectively High performance liquid chromatograph, record for test sample chromatogram ( figure 1 ) and standard chromatograms ( figure 2 ), obtained by calculating the peak area according to t...
Embodiment 2
[0057] Embodiment 2, the assay method of arginine content in Mingfule
[0058] The mobile phase described in Example 2 is 0.05 mol / L potassium dihydrogen phosphate solution: acetonitrile = 50: 50, and other parameters and operations are as shown in Example 1.
[0059] Determination steps are as shown in embodiment 1, measurement result image 3 shown. It can be seen from the figure that the ratio of potassium dihydrogen phosphate solution to acetonitrile is changed, and the peak eluting time is shortened, but there is a tailing phenomenon, and there are more impurity peaks.
Embodiment 3
[0060] Embodiment 3, the assay method of arginine content in Mingfule
[0061] The mobile phase described in Example 3 is 0.05 mol / L potassium dihydrogen phosphate solution:acetonitrile=60:80, and other parameters and operations are as shown in Example 1.
[0062] Determination steps are as shown in embodiment 1, and measurement result is as follows Figure 4 shown. It can be seen from the figure that changing the ratio of potassium dihydrogen phosphate solution to acetonitrile leads to poor peak shape, tailing phenomenon, poor separation effect and more impurity peaks.
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