Development of an HPTLC-based diagnostic method for invasive aspergillosis
Article first published online: 23 DEC 2009
Copyright © 2009 John Wiley & Sons, Ltd.
Volume 24, Issue 8, pages 887–892, August 2010
How to Cite
Puri, A., Ahmad, A. and Panda, B. P. (2010), Development of an HPTLC-based diagnostic method for invasive aspergillosis. Biomed. Chromatogr., 24: 887–892. doi: 10.1002/bmc.1382
- Issue published online: 14 JUL 2010
- Article first published online: 23 DEC 2009
- Manuscript Accepted: 3 NOV 2009
- Manuscript Received: 24 JUL 2009
- diagnostic methods;
- invasive aspergillosis;
A rapid, sensitive and specific high-performance thin-layer chromatographic (HPTLC) method was developed and validated for determination of gliotoxin in Aspergillus infected immunocompromised patients with invasive aspergillosis (IA). Densitometric analysis of gliotoxin was carried out in the absorbance mode at 254 nm after single-step extraction with chloroform. The method uses TLC aluminum plates pre-coated with silica gel 60F-254 as a stationary phase and toluene–isoamyl alcohol–methanol (10:0.5:0.5, v/v/v) as mobile phase, which gives compact spot of gliotoxin (Rf = 0.51). The calibration curve was linear (r2 ≥ 0.994) between peak area and concentration in the tested range of 100–1000 ng spot−1 with minimum detectable range 0.025 ng μ−1 of serum sample. The mean ± SD value of slope and intercept of the standard chromatogram of gliotoxin were found to be 523.2 ± 1.555635 and 915.8 ± 30.68843, respectively. The developed method is simple, rapid, precise and less costly than earlier diagnostic methods, and different serum samples can be run on a single TLC plate for comparative analysis. The proposed method can be used to analyze gliotoxin in patient serum for easy, rapid and cost-effective diagnosis of IA. Copyright © 2009 John Wiley & Sons, Ltd.