Using Aqueous Normal Phase - Microsolv

Discussion of Chromatograms. Chromatograms A & B. Peak 1 (Metformin) eluted with longer retention time as the concentration of Acetonitrile was increased.
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HPLC

TM

TM

Using Aqueous Normal Phase (ANP) and Aqueous Reverse Phase (ARP) on One TYPE-C™ Based HPLC Column Swap Peak Order… Can Your HPLC Column Do This? Method Conditions

Retention Map

Mobile Phase

Chromatograms A & B Solvent A: Aqueous (0.05% H3P04) Solvent B: Acetonitrile

Column

Cogent UDC-Cholesterol™

Cat. No.

69069-75R

Dimensions

75 x 4.6mm id

100 90

70 60

100

pH 2.18

50 40

90

Standards 1 — Uracil 2 — Pyridine 3 — Phenol

30 20 10

0

1

2

3

4

5

6

7

8

9

10

11

80

% ACN

% ACN

80

70 60 50 40 30 20

TIME IN MINUTES

Chromatogram A: Mobile Phase A -20% B -80%

2

Flow Rate

ml/min pH1.0 2.18

Detection:

UV, 240nm

10

0

SOLVENT FRONT

Standards 1 — Uracil 2 — Pyridine 3 — Phenol

UDC Cholesterol

1

2

3

4

5

6

7

8

9

10

11

RetentionTIME Map: IN MINUTES Useful tool, for quickly developing methods in HPLC: Here, Phenol exhibits a typical reverse phase plot (ARP) while both Uracil and Pyridine are showing normal phase characteristics in aqueous conditions (ANP).

Discussion of Chromatograms 1

1

Chromatograms A & B Peak 1 (Metformin) eluted with longer retention time as the concentration of Acetonitrile was increased. During the same run, Peak 2 (Glyburide) elutes with shorter retention time as concentrations of Acetonitrile are increased.

4

Chromatogram B: 1 SOLVENT FRONT

APPLICATION NOTES

No. 1 of Series D

1

2

Mobile Phase A -50% B -50%

4

Peaks 1: Metformin Log P Minus 2.6 2: Glyburide Log P Plus 4.79

Implications and Advantages Metformin is a very polar pharmaceutical compound which, chromatographically, is very similar to Pyridine (see Retention Map above). Glyburide, a neutral pharmaceutical ingredient is often formulated with Metformin, which is so polar that it elutes on the solvent front on other C18, Cyano, Amino, Phenyl and F5 columns. To quantify these compounds, two runs with different methods were previously necessary. Uses By using one UDC-Cholesterol™ HPLC Column, you can isocratically achieve separation and get confirmation of your peaks with the column’s orthogonal selectivity. Simply modify the mobile phase as shown here to save time and lab resources.

MTC • MicroSolv Technology Corporation 1 Industrial Way West, Bldg E, Eatontown, NJ 07724 USA Ph. 1-732-578-1777 • Fx. 1-732-578-9777 www.mtc-usa.com Email: [email protected] 03-H-09-D1