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HPLC Application Note: 68-AB

Separation of Cephalosporins on HALO Penta-HILIC and HALO HILIC PEAK IDENTITIES: 1

1. 2. 3. 4. 5. 6. 7. 8. 9. 10.

G0050

Absorbance @ 254 nm

HALO Penta-HILIC 2 3 4

5

1

8

9

5

10

STRUCTUR G0051

HALO HILIC

1 2,3 4

0

6 7

6 8 7

9

10

2

3

4

5

6

7

8

9

Time (min.)

Cephalothin Cefoxitin Cefotaxime Cefazolin Cefaclor Cephalexin Cephradine Cefadroxil Ceftazidime Cephalosporin C

10

STRUCTURES:

TEST CONDITIONS: Columns: 2.1 x 150 mm Part Numbers: HALO Penta-HILIC: 92812-705 HALO HILIC: 92812-701 Mobile Phase: A= 95/5 ACN/H2O with 5 mM NH4formate, pH=3.0 (adj.); B= 50/50 ACN/H2O with 5 mM NH4formate, pH=3.0 (adj.) Gradient: HALO Penta-HILIC: 85-65% B in 10 min. HALO HILIC: 85-70% B in 10 min. Flow Rate: 0.5 mL/min. Pressure: 195 bar on HALO Penta-HILIC 163 bar on HALO HILIC Temperature: 30° C Detection: UV 254 nm, VWD Injection Volume: 0.5 μL Sample Solvent: 50/50 ACN/water Flow Cell: 5 μL semi-micro LC System: Agilent 1100

Cephalothin

Cefaclor

Cefadroxil

Cephalexin

Ceftazidime

Cephradine

Cephalosporin C

Cefoxitin

Cefotaxime

The HALO Penta-HILIC phase offers an alternate selectivity for the separation of cephalosporins in HILIC mode. Cefazolin

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Cephalosporins on Penta-HILIC and HILIC