Chromatographic Conditions

S. No.

Analytical Column

Mobile phase composition

Flow

rate

Retention

time (min)

Result

1)

Zorbex SB phenyl, L11 (4.6 mm × 250 mm) 5 µm

pH 2.8 phosphate buffer, methanol and acetonitrile at a ratio of (50:30:20)

0.9 ml/min

11.99 min & 4.18 min

Improper peak separation and more RT

2)

Zorbex SB phenyl, L11 (4.6 mm × 250 mm) 5 µm

pH 2.8 phosphate buffer, methanol and acetonitrile at a ratio of (50:30:20)

1.0 ml/min

10.81 min & 3.77 min

More RT but pack separation, theatrical plate and trailing factor was good.

3)

Zorbex SB phenyl, L11 (4.6 mm × 250 mm) 5 µm

pH 2.8 phosphate buffer, methanol and acetonitrile at a ratio of (50:30:20)

1.1 ml/min

9.84 min & 3.43 min

Improper peak separation and more RT

4)

Zorbex SB phenyl, L11 (4.6 mm × 250 mm) 5 µm

pH 2.7 phosphate buffer, methanol and acetonitrile at a ratio of (50:30:20)

1.0 ml/min

10.74 min & 3.76 min

Improper peak separation and more RT

5)

Zorbex SB phenyl, L11 (4.6 mm × 250 mm) 5 µm

pH 2.9 phosphate buffer, methanol and acetonitrile at a ratio of (50:30:20)

1.0 ml/min

10.36 min & 3.74 min

Improper peak separation and more RT

6)

Zorbex SB phenyl, L11 (4.6 mm × 250 mm) 5 µm

48:52 Mobile Phase Ratio

1.0 ml/min

8.78 min & 3.58 min

Improper peak separation, more tailing and less plate count

7)

Zorbex SB phenyl, L11 (4.6 mm × 250 mm) 5 µm

52:48 Mobile Phase Ratio

1.0 ml/min

12.13 min & 3.89 min

Improper peak separation and more RT

8)

Phenomenex C18 ODS (4.6 × 150 mm) 5 µm

pH 2.8 phosphate buffer, methanol and acetonitrile at a ratio of (50:30:20)

1.0 ml/min

7.8 min & 3.42 min

Improper peak separation, more tailing and less plate count

9)

Inertsil C18 (15 cm × 0.46 cm) 5 µm

pH 2.8 phosphate buffer, methanol and acetonitrile at a ratio of (50:30:20)

1.0 ml/min

14.2 min & 4.9 min

Improper peak separation and more RT