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1.
TLS Spectrometry & Detection in Liquid Chromatography, FIA and Microfluidic Systems
Mladen Franko, invited lecture at foreign university

Abstract: This presentation provides theoretical backround for thermal lens spectrometry (TLS) and TLS microscopy (TLM) and describes the basic instrumentation for TLS and TLM measurements. Presented applications of TLS and TLM include detection in HPLC (carotenoids, bilirubin, biliverdin), FIA (alergens, Ag, Ag+, phytoplankton and cyanobacterial pigments, organophospate pesticides) and TLM detection in microfluidic systems (Cr(VI), microcystin, NGAL, study of diffusion proceses)
Keywords: Spekrometry TLS, TLM, HPLC, FIA, microfluidics, bilirubin, biliverdin, carotenoids, alergens, mikrocystin, pigments, organophosphate pesticides, NGAL, Ag, Cr(VI), diffusion
Published in RUNG: 16.09.2016; Views: 5430; Downloads: 3
.pdf Full text (35,40 MB)

2.
Application of high-performance liquid chromatography combined with ultra-sensitive thermal lens spectrometric detection for simultaneous biliverdin and bilirubin assessment at trace levels in human serum
Mitja Martelanc, Lovro Žiberna, Sabina Passamonti, Mladen Franko, 2016, original scientific article

Abstract: We present the applicability of a new ultra sensitive analytical method for the simultaneous determination of biliverdin and bilirubin in human serum. The method comprises isocratic reversed-phase(RP) C18 high-performance liquid chromatography (HPLC) and thermal lens spectrometric detection (TLS)based on excitation by a krypton laser emission line at 407 nm. This method enables the separation of IX-α biliverdin and IX-α bilirubin in 11 min. with limit of detection (LOD) and limit of quantitation (LOQ) for biliverdin of 1.2 nM and 3 nM, and 1nM and 2.8 nM for bilirubin,respectively.In addition, a step-gradient elution was set up, by changing the mobile phase composition, in order to further enhance the sensitivity for bilirubin determination with LOD and LOQ of 0.5 nM and 1.5 nM, respectively. In parallel, an isocraticHPLC-DAD method was developed for benchmarking against HPLC-TLS methods. The LOD and LOQ forbiliverdin were 6 nM and 18 nM, and 2.5 nM and 8nM for bilirubin,respectively. Additionally, both isocratic methods were applied for measuring biliverdin and free bilirubin in human serum samples (from 2 male and 2 female healthy donors). Combining isocratic HPLC method with TLS detector was crucial for first ever biliverdin determination in serum together with simultaneous free bilirubin determination. We showed for the first time the concentration ratio of free bilirubin versus unbound biliverdin in human serum samples.
Keywords: Bilirubin, biliverdin, serum, HPLC, TLS
Published in RUNG: 29.03.2016; Views: 5805; Downloads: 0
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