Monolithic chromatography on conjoint liquid chromatography columns for speciation of platinum-based chemotherapeutics in serum of cancer patients |
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| Affiliation: | 1. Department of Environmental Sciences, Jožef Stefan Institute, Jamova 39, 1000 Ljubljana, Slovenia;2. Jožef Stefan International Postgraduate School, Jamova 39, 1000 Ljubljana, Slovenia;3. Department of Experimental Oncology, Institute of Oncology Ljubljana, Zaloška cesta 2, 1000 Ljubljana, Slovenia;4. Department of Medical Oncology, Institute of Oncology Ljubljana, Zaloška cesta 2, 1000 Ljubljana, Slovenia;1. Chemistry department, Faculty of Science, Islamic University of Gaza, Palestine;2. Faculty of Medicine, Islamic University of Gaza, Palestine;1. School of Health and Social Care, Teesside University, Middlesbrough, UK;2. Centre for Oral Health Research, School of Dental Sciences, Newcastle University, Newcastle upon Tyne, UK;1. MOE Key laboratory of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631, China;2. College of Environment and Chemical Engineering, Dalian University, Dalian, Liaoning 116622, China;1. Federal Research and Clinical Center of Physical Chemical Medicine, 1-a Malaya Pirogovskaya, Moscow, 119435, Russia;2. Lebedev Physical Institute, 53 Leninskij Prospekt, Moscow, 119991, Russia;3. Moscow Research Institute of Psychiatry, 3 Poteshnaya, Moscow, 107076, Russia |
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| Abstract: | BackgroundMonolithic chromatography using convective interaction media (CIM) disks or columns can be used in the separation step of speciation analysis. When different monolithic disks are placed in one housing, forming conjoint liquid chromatography (CLC) monolithic column, two-dimensional separation is achieved in a single chromatographic run.MethodsHere, we assembled low-pressure (maximum 50 bar) CLC monolithic column, which consists of two 0.34 mL shallow CIM monolithic disks and high-pressure CLC column (maximum 150 bar) from 0.1 mL analytical high performance short bed CIMac monolithic disks. Both the CLC columns constructed from affinity Protein G and weak anion exchange diethylamine (DEAE) disks, were applied for the speciation of cisplatin, oxaliplatin and carboplatin in spiked standard serum proteins, spiked human serum and serum of cancer patients. The analytical performances of the CLC columns used were evaluated by comparing their robustness, selectivity, repeatability and reproducibility. The separated serum proteins were detected on-line by ultraviolet (UV) and eluted Pt species by inductively coupled plasma mass spectrometry (ICP-MS). For accurate quantification of the separated Pt species (unbound Pt-based chemotherapeutic from species associated to transferrin (Tf), human serum albumin (HSA) and Immunoglobulin G (IgG)), post column isotope dilution (ID)-ICP-MS was used.ResultsThe data from analyses showed that both tested CLC monolithic columns gave statistically comparable results, with the low-pressure CLC column exhibiting better resolving power and robustness. It also enables more effective cleaning of monolithic disks and to analyse larger series of serum samples than the high-pressure CLC column. Analyses of serum samples of cancer patients treated with cisplatin or carboplatin showed that Pt-chemotherapeutics were bound preferentially to HSA (around 80%). The portion of unbound Pt in general did not exceed 2%, up to 5% of Pt was associated with Tf and approximately 20% with IgG. Column recoveries, calculated as a ratio between the sum of concentrations of Pt species eluted and concentration of total Pt in serum samples, were close to 100%.ConclusionsLow-pressure CLC column exhibited greater potential than high-pressure CLC column, and can be thus recommended for its intended use in speciation analysis of metal-based biomolecules. |
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| Keywords: | Low-pressure and high-pressure conjoint liquid chromatography Anion-exchange and affinity monolithic disks Inductively coupled plasma mass spectrometry Pt-based chemotherapeutics Serum of cancer patients |
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