Unambiguous determination of farnesol and tyrosol in vaginal fluid using fast and sensitive UHPLC-MS/MS method
Autor: | Michele Protti, Frantisek Svec, Laura Mercolini, Vladimír Buchta, Hana Vlčková, Ondřej Jung, Lucie Nováková, Veronika Pilařová |
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Přispěvatelé: | Pilarova V., Kocova Vlckova H., Jung O., Protti M., Buchta V., Mercolini L., Svec F., Novakova L. |
Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
Adult
Candida albican Microextraction Sorbent 02 engineering and technology Tandem mass spectrometry 01 natural sciences Biochemistry Analytical Chemistry chemistry.chemical_compound Young Adult Limit of Detection Tandem Mass Spectrometry Candida albicans Tyrosol Protein precipitation Humans Sample preparation Chromatography High Pressure Liquid Chromatography Chemistry Elution 010401 analytical chemistry Extraction (chemistry) Selected reaction monitoring Middle Aged Phenylethyl Alcohol 021001 nanoscience & nanotechnology Farnesol 0104 chemical sciences Quorum sensing UHPLC-MS/MS Vagina Female 0210 nano-technology |
Popis: | The new ultra-high performance liquid chromatography method with tandem mass spectrometry detection (UHPLC-MS/MS) has been optimized to allow fast, selective, and high-throughput analysis of two Candida albicans quorum sensing molecules (QSM), farnesol and tyrosol. The problem of the presence of the interference in the samples and system was successfully solved by careful optimization of chromatographic conditions. Charged hybrid stationary phase modified with pentafluorophenyl group and optimized gradient elution provided adequate separation selectivity and peak shapes. The impurity was identified as dibutyl phthalate and had the same m/z ions as farnesol leading to an important interference on selected reaction monitoring channel. Two different types of biological matrices originating from vaginal fluid, supernatant and sediment, were analysed. Micro-solid phase extraction in pipette tips was optimized for the selective isolation of QSM from the supernatant. The insufficient retention of farnesol on the extraction sorbent was improved when 1% of organic solvent was added prior to extraction, while the retention of tyrosol was only possible when using combined C8 and polymer sorbent type. Strong retention of farnesol had to be solved by increasing elution solvent strength and volume up to 600μL. However, this approach did not allow the pretreatment of sediment samples due to the sorbent clogging. Therefore, our previously developed protein precipitation method was modified and validated to analyse the sediments. New developed UHPLC-MS/MS method provided suitable accuracy and precision for the determination of QSM in vaginal fluid while using only 50μL sample volume and two different sample preparation methods. |
Databáze: | OpenAIRE |
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