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Table 1 Effect of experimental parameters on the separation of EBS-Zn (ΙΙ) complex

From: Validated stability indicating liquid chromatographic determination of ebastine in pharmaceuticals after pre column derivatization: Application to tablets and content uniformity testing

Parameter

K'

N

HETP

Rs

Peak area ratio

Buffer type and pH

Britton Robinson buffer pH

 

3

3.41

1036

0.0145

2.73

0.726

3.5

3.92

1034

0.0145

3.58

0.743

4

5.53

2706

0.0055

5.012

0.784

4.5

5.13

2698

0.0055

5.012

0.784

5

3.84

1507

0.0099

4.177

0.752

5.5

3.66

769

0.0195

3.166

0.688

6

3.76

258

0.0581

3.133

0.564

Phosphate buffer pH 4.2

4.26

1479

0.0101

2.72

0.366

Acetate buffer pH 4.2

4.06

1027

0.0146

1.271

0.521

Ratio of aqueous: organic phase(acetonitril: methanol; 1:4)

45:55

4.02

837

0.0179

1.088

0.29

40:60

4.25

1034

0.0145

4.15

0.45

35:65

5.53

2706

0.0055

5.012

0.784

30:70

5.52

2687

0.0056

5.009

0.783

Ratio of acetonitril: methanol

1:4

5.53

2706

0.0055

5.012

0.784

2:3

3.52

1210

0.0124

3.64

0.645

3:2

2.73

2205

0.0068

4.45

0.429

4:1

4.103

1393

0.0108

4.41

0.235

Acetonitril only

5.08

1347

0.0111

4.745

0.198

Methanol only

4.938

1817

0.0083

4.371

0.349

Ionic strength of BRb (Molar)

0.0075

4.256

911

0.0165

3.714

0.48

0.01

5.23

1895

0.0079

4.015

0.69

0.015

5.53

2706

0.0055

5.012

0.784

0.02

5.52

2716

0.0053

5.009

0.782

0.05

5.55

2720

0.0052

5.011

0.783

0.07

4.62

1816

0.0083

4.127

0.642

0.1

4.67

1173

0.0128

4.143

0.513

Flow rate (mL/minute)

0.6

4.348

466

0.0322

2.11

0.92

0.7

4.291

1013

0.0148

2.64

0.84

0.8

4.342

1070

0.0141

2.69

0.75

1

5.53

2706

0.0055

5.012

0.784

1.1

5.26

1982

0.0076

4.857

0.662

1.2

4.74

1486

0.0101

4.33

0.679

1.3

4.25

1057

0.0142

3.21

0.54

1.4

4.12

1013

0.0148

2.94

0.49

1.5

3.97

925

0.0162

2.76

0.33

  1. Where:
  2. K' is the capacity factor; K'= tR - tM/tM
  3. N is the number of theoretical plates;
  4. HETP is the height equivalent to a theoretical plate; HETP = L/N
  5. Rs is the resolution factor;
  6. (t R is the retention time of the complex, t M is the time from injection to the appearance of the solvent front, w 1/2 is the peak width at half height, L is the length of the column, w R and w M are the peak width at base line for the complex and the solvent front respectively)