Page 10 - LC-SFC_Pharma_Brochure

4-7. Determining Analytical Conditions for Simultaneous Analysis

The analytical conditions optimized in Step 2 for the ultra high column, and gradient program described in Step 1 and Step 2, we
performance LC simultaneous analysis of the 13 cephem antibiot- were able to establish analytical conditions that permitted the
ic substances are shown in Table 4, and their respective chromato- completion of one cycle of simultaneous analysis of 13 cephem
grams are shown in Fig. 12. Due to optimization of the mobile phase, antibiotics within 9 minutes.
mAU
7
400

Peaks
9
1. Cefsulodin
300
2. Cefadroxil
3 3. Cephapirin
4
4. Cefaclor
5. Cephalexin
6. Cephradine
200 8 12 13 7. Cefotaxime
8. Cefazolin
12 5 6 9. Cefuroxime
11
10. Cefmetazole
11. Cefoxitin
10
100 12. Cefoperazone
13. Cephalothin
0
0 0.5 1.0 1.5 2.0 2.5 3.0 min
Fig. 12 Simultaneous Analysis of 13 Cephem Antibiotic Substances

Table 4 Analytical Conditions for Simultaneous Analysis of 13 Cephem Antibiotics
Mobile phase :(A) Sodium phosphate buffer solution (pH 2.6)
:(B) Acetonitrile
Column :Kinetex XB-C18 (50 mmL. × 3.0 mmI.D., 2.6 µm)
Time program :B Conc. 5% (0 min) 65% (5 min) 90% (5.01−7 min)
Flow rate :1.0 mL/min
Injection volume :5 µL
Column temperature :40°C
Detection wavelength :260 nm (SPD-M20A)

(Note) Kinetex is a trademark or registered trademark of Phenomenex, Inc. in the U.S. and other countries.
For details regarding this product, please contact the Shimadzu GLC. – Phenomenex Support Center (e-mail psc@glc.shimadzu.co.jp)
or your Shimadzu representative

First Edition: December, 2012

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