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Analysis of flavonoids in beer by LC-MS
Měřínská, Radana ; Kočí, Radka (referee) ; Márová, Ivana (advisor)
Presented diploma thesis is focused on analysis of phenolic compounds in various types of beers. The aim of this work was to find possible differences among several types of beers and also between Czech beers and beers of foreign production. Theoretical part contains detailed description of beer technology, review of important analytical methods for determination of phenolic compounds and total antioxidant activity. The main attention is focused on instrumental technique LC/MS. In the experimental part values of total phenolics and total flavonoids and results of antioxidant status in analyzed beers were determined spectrophotometrically. Basic brewing characteristics were determined by pycnometry. On-line liquid chromatography with photo diode array detection and mass spectrometry detection was used for identification and quantification of individual phenolic compounds. Spectrophotometric analysis of phenolic and flavonoid levels seems to be less specific, that is consistent with findings in accessible literature. It was proved, that iso- bitter compounds exhibit about 65.44 – 90.93 % of the total bitter substances. These components are responsible for the main part of beer bitterness. The HPLC/PDA/ESI-MS was necessary to use for determination of characteristic phenolic compounds in individual types of beer. Within our measurement three columns were tested, the most favourable results were obtained using the column Restek C18. When compared with foreing beers, in Czech beers higher level of majority of phenolic compounds was detected and specific distribution of individual derivatives was found as well.
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Study of phenolic compounds in some biologic materials using method LC/MS
Měřínská, Radana ; Zemanová, Jana (referee) ; Kočí, Radka (advisor)
Presented bachelor thesis was focused on suitable methods for analysis of phenolics, especially modern instrumental methods were studied. In experimental part some standard phenolic compounds were analysed using combined technique HPLC/ESI-MS with off-line and/or on-line detection. Mass spectra of individual stadards and/or mixtures were determined in MS full scan and/or MS/MS full scan mode. Ionisation of chosen standard compounds was done in pozitive and/or negative mode. Parent ions of followed compounds were identified: reserpine (PI 609.9, NI 607.7), acid chlorogenic (NI 353), quercetin (PI 303.5, NI 301.5), rutin (PI 611, NI 609), morin (PI 303.1, NI 301.1). Typical fragments of studied standards were produced using MS/MS full scan mode. By fragmentation characteristic product ions were identified: acid chlorogenic (NI 191), quercetin (PI: 229.1 and 257.1; NI: 179.0 and 151.0), rutin (in MS/MS NI: 301; in MS3 NI: 179, 151.1), morin (PI: 285.1 a 257.1; NI: 283.2, 273.2, 257.1, 229.1). Optimal conditions of phenolic HPLC separation were determined including gradient elution and Restek column use. In conclusion on-line HPLC/MS detection using negative ionization was observed as the best configuration for phenolic analysis.
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Analysis of flavonoids in beer by LC-MS
Měřínská, Radana ; Kočí, Radka (referee) ; Márová, Ivana (advisor)
Presented diploma thesis is focused on analysis of phenolic compounds in various types of beers. The aim of this work was to find possible differences among several types of beers and also between Czech beers and beers of foreign production. Theoretical part contains detailed description of beer technology, review of important analytical methods for determination of phenolic compounds and total antioxidant activity. The main attention is focused on instrumental technique LC/MS. In the experimental part values of total phenolics and total flavonoids and results of antioxidant status in analyzed beers were determined spectrophotometrically. Basic brewing characteristics were determined by pycnometry. On-line liquid chromatography with photo diode array detection and mass spectrometry detection was used for identification and quantification of individual phenolic compounds. Spectrophotometric analysis of phenolic and flavonoid levels seems to be less specific, that is consistent with findings in accessible literature. It was proved, that iso- bitter compounds exhibit about 65.44 – 90.93 % of the total bitter substances. These components are responsible for the main part of beer bitterness. The HPLC/PDA/ESI-MS was necessary to use for determination of characteristic phenolic compounds in individual types of beer. Within our measurement three columns were tested, the most favourable results were obtained using the column Restek C18. When compared with foreing beers, in Czech beers higher level of majority of phenolic compounds was detected and specific distribution of individual derivatives was found as well.
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Study of phenolic compounds in some biologic materials using method LC/MS
Měřínská, Radana ; Zemanová, Jana (referee) ; Kočí, Radka (advisor)
Presented bachelor thesis was focused on suitable methods for analysis of phenolics, especially modern instrumental methods were studied. In experimental part some standard phenolic compounds were analysed using combined technique HPLC/ESI-MS with off-line and/or on-line detection. Mass spectra of individual stadards and/or mixtures were determined in MS full scan and/or MS/MS full scan mode. Ionisation of chosen standard compounds was done in pozitive and/or negative mode. Parent ions of followed compounds were identified: reserpine (PI 609.9, NI 607.7), acid chlorogenic (NI 353), quercetin (PI 303.5, NI 301.5), rutin (PI 611, NI 609), morin (PI 303.1, NI 301.1). Typical fragments of studied standards were produced using MS/MS full scan mode. By fragmentation characteristic product ions were identified: acid chlorogenic (NI 191), quercetin (PI: 229.1 and 257.1; NI: 179.0 and 151.0), rutin (in MS/MS NI: 301; in MS3 NI: 179, 151.1), morin (PI: 285.1 a 257.1; NI: 283.2, 273.2, 257.1, 229.1). Optimal conditions of phenolic HPLC separation were determined including gradient elution and Restek column use. In conclusion on-line HPLC/MS detection using negative ionization was observed as the best configuration for phenolic analysis.
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