Sr .No.

Parameter

Level

RT*
(min)

As USP**

Res USP***

1

Flow rate (mL/min)

0.9

7.18

1.21

2.16

1.0

6.41

1.18

2.18

1.1

5.92

1.16

2.06

2

pH of the buffer

3.0

6.41

0.97

2.23

3.3

6.5

1.16

2.15

3

Column temperature (oC)

25

6.41

1.18

2.18

31

6.12

1.17

2.12

37

6.1

1.2

2.02

4

Composition of solvent B (%)

49

5.97

1.2

2.01

50

6.41

1.18

2.18

51

6.3

1.17

2.08

5

Injection volume (µL)

20

6.41

1.18

2.18

50

6.41

1.14

1.62

6

Wavelength λ max-(nm)

275

6.44

1.22

2.1

280

6.41

1.18

2.18

285

6.48

1.18

2.14

*RT- Retention Time, **As USP- Asymmetry USP, *** Res USP- Resolution USP
Table 4: Results of the robustness of the method.