Trials

Chromatographic conditions

S. No.

Analytical Column

Mobile phase composition

Flow rate

Retention time (min)

Result

1

Inertsil C18

(15 cm × 0.46 cm) 5 µm

Water: Methanol

(50:50% v/v)

0.8 ml/min

7.553

Improper peak separation and more RT.

2

Zodiac C18 (4.6 × 150 mm) 5 µm

ACN: MeOH

(40:60%) v/v

0.9 ml/min

2.508

Improper peak separation and less plate count.

3

Develosil C18 (4.6 × 150 mm) 5 µm

ACN: 0.1% OPA (pH-3.6)

(30:70) v/v

1.0 ml/min

3.213

Improper peak separation and less plate count.

4

Phenomenex C18 ODS

(4.6 × 150 mm) 5 µm

ACN: Ammonium buffer (0.1 M)

(45:55% v/v)

1.0 ml/min

3.213

Improper peak separation, more tailing and less plate count.

5

Symmetry C18 ODS (4.6 × 150 mm) 5 µm

MeOH: Phosphate buffer (0.01 M)

(40:60%) v/v

1.0 ml/min

4.161

Improper peak separation and peak broadening.

6

Symmetry C18 ODS (4.6 × 150 mm) 5 µm

MeOH: Phosphate buffer (0.05 M)

(20:80%) v/v

1.0 ml/min

4.396

Improper peak separation, broad peak and peak tailing.