Xtimate Polar RP-Polymer Impurity-Preparation and Purification-20240218.docx

Test Report

• 1. Experimental Procedure

1.1. Method Reproduction
Weigh 5 mg of the sample, add 5 ml of pure water to dissolve it, filter into an injection vial, and analyze according to the following chromatographic analysis method.

Chromatography conditions:

The analysis spectrum is shown in Figure 2:

Figure 2: Method Reproduction Chromatogram

Conclusion: By comparing with Figure 1, it can be determined that 3.728 minutes is the target impurity.

• 1.1.1. Sample Preparation

Weigh an appropriate amount of the sample, add pure water and acetonitrile to dissolve and dilute it, filter to make its concentration 10 mg/ml, and prepare the liquid phase conditions as follows:

The preparation spectrum is shown in Figure 4:

Figure 4: Crude Product Preparation Diagram

After preparation, collect the target fraction, pre-freeze it with dry ice and ethanol, then lyophilize using a freeze dryer.

• 1.1.2. Impurity Analysis

Analyze the lyophilized impurities from 1.1.1 using liquid chromatography under the same conditions as in "Step 1.1." The analysis chromatogram is shown in Figure 5.

Figure 5: Prepared Solution Analysis Chromatogram

The blank analysis chromatogram is shown in Figure 6.

Conclusion: From the figure, it can be seen that after removing the blank, the area normalization content of the prepared solution is 100% (214 nm), and the purity meets the customer's requirements.

• 2. Conclusion 

Using Welch Xtimate Polar RP (10*250 mm, 5 µm) under these chromatographic conditions, with a single injection sample amount of 5 mg, the purity of the collected target substance is 100% (214 nm), and the preparation results meet the customer's requirements.