No

 

Chromatographic condition

Parameters of absorption peaks

Estimate of the results

1

30%B+70%A changed to 45%B+55%A in 10 min

retention time: 1.946

resolution: 1.210

symmetry factor: 0.951

theoretical plate: 3408

The retention time is short, and a certain degree of separation is obtained.

2

27%B+73%A changed to 45%B+55%A in 10 min

retention time: 3.051

resolution: 1.365

symmetry factor: 0.996

theoretical plate: 4068

The retention time is short,and a certain degree of separation is obtained.

3

20%B+80%A changed to 50%B+50%A in 10 min

retention time: 6.403

resolution: 1.610

symmetry factor: 0.993

theoretical plate: 4745

The retention time is proper, and good resolution is obtained.

4

30%B+70%A , isocratic elution, 0-15 min

The sample is remained in the column.

The sample is retained in 

the column

5

35%B+65%A, isocratic elution, 0-15 min,

retention time: 2.152

resolution: 1.214

symmetry factor: 0.966

theoretical plate: 2055

The retention time is too short, and the resolution is not good.

Table 1: The chromatographic conditions and results. A is 0.05% TFA aqueous solution and B is acetonitrile. The sample which was injected into the HPLC system in this study is CMS (0.5 mg/mL). The parameters are the values from measurements of CMS E1.