8-Hydroxydeoxyguanosine detection kit and its application
A technology of hydroxydeoxyguanosine and a kit, which is applied in the detection field of 8-hydroxydeoxyguanosine, can solve the problems of higher detection value than the real value, poor specificity of immunochemical method, and simultaneous washing out of interfering substances, etc. Cleaning liquid path and column, good day-to-day precision, and good recovery of standard addition
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Embodiment 1
[0093] Example 1 Example 1 Take NaH 2 PO 4 KH 2 PO 4 Comparative study for mobile phase detection
[0094] Mobile phase 1: 50mmol / l Ph3.5 NAH 2 PO 4 Solution: methanol = 90%: 10%.
[0095] Mobile phase 2: 50mmol / L pH 3.5 kH 2 PO 4 Solution: methanol = 90%: 10%.
[0096] The injection volume was 50 μl, the flow rate was 1.0 mL / min, the column temperature was 30 ° C, and the wavelength was 300 nm.
[0097] The 8-hydroxy deoxycoside standard control solution was prepared with a concentration of 10 μg / ml.
[0098] The results detected by mobile phase 2 figure 2 . The results detected by mobile phase 1 image 3 Indicated.
[0099] As can be seen from the figure, with kH 2 PO 4 When the flow of the phase, the peak peak of 8-OHDG at 13-16 minutes has a shoulder, and the chromatographic separation does not meet the test requirements. Nah 2 PO 4 In the case of flowing a phase, the peak peak shape of 8-OHDG at 14-16 minutes is good, in line with the test requirements.
Embodiment 2
[0100] Example 2 Selection of flow phase solvent
[0101] Mobile phase A is Nah 2 PO 4 The solution, the mobile phase B is methanol or acetonitrile.
[0102] Use 50 mmol / l pH 3.5 NAH 2 PO 4 Solution: Acetonitrile = 90%: 10% chromatographic conditions measurable 8-hydroxy deoxymide control, acetonitrile for the separation of the 8-OHDG target sample peak and solvent peak Figure 4 .
[0103] Use 50 mmol / l pH 3.5 NAH 2 PO 4 Solution: Methanol = 90%: 10% chromatographic conditions measured 8-hydroxy deoxymide control, methanol to see the separation of the 8-OHDG target sample peak and solvent peak Figure 5 .
[0104] by Figure 4 with Figure 5 It can be seen that the separation of acetonitrile and methanol can be used as the flow phase solvent of the present invention.
[0105] Considering that methanol is lower than the use of acetonitrile, it is more conducive to the application of clinical large-area applications, so that methanol is preferably selected as organic phases.
Embodiment 3
[0106] Example 3 Selection of column temperature
[0107] Mobile phase A is 50 mmol / l pH 3.5 Nah 2 PO 4 The solution, the mobile phase B is methanol, and the mobile phase A: the mobile phase B is 9: 1, the injection volume is 50 μl, and the flow rate is 1.0 mL / min.
[0108] The 8-hydroxy deoxycine control was added to a healthy human urine to give a concentration of 20 μg / ml, comparison 10 ° C, 20 ° C, 30 ° C different column temperature conditions under different column temperature conditions.
[0109] Table 2 Reserved time for reference under different column temperature conditions
[0110]
[0111] From the above table, the column temperature is 20-30 ° C, and the 8-hydroxyl deoxycoside component peak is good, and can be separated from other impurities peaks. In view of 30 ° C, the peak peak peak peak peak peak is relatively faster, which is advantageous for increasing the efficiency of clinical mass sample detection, and thus the preferred column temperature is 30 ° C....
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