HPLC DETERMINATION OF SOME ANTIBIOTICS
Virginia Coman1, Veronica Avram1, Loredana Soran1, Rodica Grecu1,
Z. Moldovan2 and Hajnalka Farkas3
“Raluca Ripan” Institute for Research in Chemistry, Department of
Chromatography, 30 Fântânele Street, Cluj-Napoca, Romania,
E-mail: v.coman@icrr.cj.edu.ro
2
National Institute for Research & Development of Isotopic and
Molecular Tehnology, 71-103 Donath Street, 400293 Cluj-Napoca,
Romania
3
“Babeş-Bolyai” University, Faculty of Chemistry and Chemical
Engineering, 11 Arany Janos Street, RO-3400 Cluj-Napoca, Romania
1
The determination of antibiotics presents some difficulties because of their thermal instability.
The HPLC method at room temperature is preferred in the last decade.
This work presents a simple HPLC method for the determination of some antibiotics (see
Table). The registering of chromatograms was performed with a high performance liquid
chromatograph JASCO – 980 (JASCO, Japan) equipped with an UV-VIS detector (190–400
nm). The samples were introduced by manual injection with Hamilton Rheodyne Syringe (50
L) in a valve of 20 L loop. To find the optimum UV wave length for these antibiotic
detection, the UV-VIS spectra registered on UNICAM UV4 spectrometer were analysed.
For the determination of mentioned antibiotics, the following experimental HPLC parameters
were established: wave length 220 nm; separation column: NUCLEOSIL 120–C18, 5,
250.4 cm; mobile phase: acetonitrile – water with ammonium acetate buffer 80 mM, in the
ratio 20:80 (v/v); flow rate of 1 mL/min.
Some antibiotics were detected also at 270 nm (cefuroxime, chloramphenicol), and 250
nm (gentamicin) in order to improve the HPLC detection limit.
The HPLC data have been processed by the BORWIN Soft. For all antibiotics the calibration
curves were registered in the concentration range of 0.2–1.0 mg/ml.
The retention time (RT), detection limit (DT) and regression factor (r) for studied antibiotics
are given in the table below:
TABLE. Retention time, detection limit and regression factor for studied antibiotics.
Nr. crt.
1
2
3
4
5
6
7
Antibiotic
Ampicillin
Cefuroxime
Lincomicin
Penicillin
Gentamicin
Oxacillin
Chloramphenicol
RT [min]
2.70
3.58
5.12
7.00
7.95
11.01
12.46
DL [g/ml]
50.0
12.5
50.0
3.1
50.0
0.8
12.5
r
0.99567
0.99972
0.99960
0.99895
0.98335
0.99573
0.99925
References
[1] V.R. Meyer, Practical High-Performance liquid Chromatography, IIIth Edition, John Wiley & Sons Ltd., Chichester, 1998.
[2] D. Do b re s c u, E. Ma n o le sc u, V. S ub ţir ică , A. Dră g a n, C. I va n, L. Do b res c u, I. A. An ca ,
Memomed 2002. (Drug Memorized Book and Pharmacotherapic Guide), VIIIth Edition, Bucharest, 2002.