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Vysoké Učení Technické V Brně Brno University of Technology VYSOKÉ UČENÍ TECHNICKÉ V BRNĚ BRNO UNIVERSITY OF TECHNOLOGY FAKULTA CHEMICKÁ ÚSTAV CHEMIE POTRAVIN A BIOTECHNOLOGIÍ FACULTY OF CHEMISTRY INSTITUTE OF FOOD SCIENCE AND BIOTECHNOLOGY CHARAKTERIZACE PLODŮ ARONIE (ARONIA MELANOCARPA) A MUCHOVNÍKU (AMELANCHIER ALNIFOLIA) DIPLOMOVÁ PRÁCE MASTER'S THESIS AUTOR PRÁCE Bc. LENKA BUTOROVÁ AUTHOR BRNO 2014 VYSOKÉ UČENÍ TECHNICKÉ V BRNĚ BRNO UNIVERSITY OF TECHNOLOGY FAKULTA CHEMICKÁ ÚSTAV CHEMIE POTRAVIN A BIOTECHNOLOGIÍ FACULTY OF CHEMISTRY INSTITUTE OF FOOD SCIENCE AND BIOTECHNOLOGY CHARAKTERIZACE PLODŮ ARONIE (ARONIA MELANOCARPA) A MUCHOVNÍKU (AMELANCHIER ALNIFOLIA) CHARACTERISATION OF CHOKEBERRY AND SASKATOON BERRY FRUITS DIPLOMOVÁ PRÁCE MASTER'S THESIS AUTOR PRÁCE Bc. LENKA BUTOROVÁ AUTHOR VEDOUCÍ PRÁCE Ing. EVA VÍTOVÁ, Ph.D. SUPERVISOR BRNO 2014 Vysoké učení technické v Brně Fakulta chemická Purkyňova 464/118, 61200 Brno 12 Zadání diplomové práce Číslo diplomové práce: FCH-DIP0817/2013 Akademický rok: 2013/2014 Ústav: Ústav chemie potravin a biotechnologií Student(ka): Bc. Lenka Butorová Studijní program: Chemie a technologie potravin (N2901) Studijní obor: Potravinářská chemie a biotechnologie (2901T010) Vedoucí práce Ing. Eva Vítová, Ph.D. Konzultanti: RNDr. Milena Vespalcová, Ph.D. Název diplomové práce: Charakterizace plodů aronie (Aronia melanocarpa) a muchovníku (Amelanchier alnifolia) Zadání diplomové práce: 1. Zpracujte literární rešerši k dané problematice 2. Zvolte parametry vhodné pro charakterizaci nutriční kvality plodů aronie a muchovníku 3. Vyberte metody vhodné pro jejich stanovení 4. Aplikujte je na vybrané vzorky 5. Zhodnoťte nutriční kvalitu plodů Termín odevzdání diplomové práce: 9.5.2014 Diplomová práce se odevzdává v děkanem stanoveném počtu exemplářů na sekretariát ústavu a v elektronické formě vedoucímu diplomové práce. Toto zadání je přílohou diplomové práce. - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - Bc. Lenka Butorová Ing. Eva Vítová, Ph.D. doc. Ing. Jiřina Omelková, CSc. Student(ka) Vedoucí práce Ředitel ústavu - - - - - - - - - - - - - - - - - - - - - - - V Brně, dne 31.1.2014 prof. Ing. Jaromír Havlica, DrSc. Děkan fakulty Tato práce vznikla za podpory projektu BiochemNet - Vytvoření sítě pro podporu spolupráce biomedicínských pracovišť a zvýšení uplatnitelnosti absolventů biochemických oborů v praxi (registrační číslo CZ.1.07/2.4.00/31.0133). Projekt je realizován v rámci Operačního programu vzdělávání pro konkurenceschopnost, který je spolufinancovaný z Evropského sociálního fondu a státního rozpočtu ČR. ABSTRAKT Diplomová práce se zabývá charakterizací plod ů aronie ( Aronia melanocarpa ) a muchovníku ( Amelanchier alnifolia ). Teoretická část práce pojednává o botanickém za řazení, chemickém složení, lé čivých účincích a použití plod ů aronie a muchovníku v potraviná řském pr ůmyslu. Dále je pozornost v ěnována p řehledu metod pro stanovení d ůležitých antioxidant ů, se zam ěř ením na stanovení polyfenolických látek, anthokyan ů, kyseliny askorbové a celkové antioxida ční aktivity. Experimentální část práce se zam ěř uje na stanovení vybraných parametr ů různých odr ůd aronie a muchovník ů sbíraných v letech 2012 a 2013 pomocí spektroskopických a chromatografických technik s důrazem na vliv rozpoušt ědla (50% ethanol, destilovaná voda, DMSO) na uvedené parametry. Konrétn ě je v práci zjiš ťován celkový obsah polyfenolických látek a anthokyanů, barevné charakteristiky pomocí systému CIE L*a*b* a schopnost vzorku redukovat železité ionty pomocí metody UV-VIS-NIR spektroskopie. Dále je kvantifikován obsah kyseliny askorbové v extraktech z plod ů aronie a muchovník ů pomocí vysokoú činné kapalinové chromatografie s využitím detektoru s diodovým polem a také celková antioxida ční aktivita metodou DPPH a ABTS s využitím elektronové paramagnetické rezonance. ABSTRACT Diploma thesis deals with the characterization of chokeberry (Aronia melanocarpa ) and saskatoon berry (Amelanchier alnifolia ) fruits. The theoretical part focuses on the botanical classification, chemical composition, curative effects and the use of chokeberry and saskatoon berry in the food industry. Further, the attention is also devoted to a review of methods for the determination of important antioxidants, specialized on the determination of polyphenols, anthocyanins, ascorbic acid and total antioxidant activity. The experimental part of the work deals with the determination of selected parameters in different varieties of chokeberry and serviceberry, harvested in 2012 and 2013 using spectroscopic and chromatographic techniques with an emphasis on effect of solvent (50% ethanol, destilled water, DMSO). Specifically total content of polyphenols and anthocyanins, color characteristics using the CIE L*a*b* system and the ability of the sample to reduce the iron are determined using the UV-VIS-NIR spectroscopy. Further ascorbic acid concentration in individual extracts is quantified by High performance liquid chromatography coupled with a diode array detector and the total antioxidant activity is characterized by electron paramagnetic resonance involving •DPPH and ABTS •+ assays. KLÍ ČOVÁ SLOVA aronie, muchovník, antioxida ční aktivita, anthokyany, vitamin C, HPLC, UV/VIS/NIR, EPR spektroskopie KEY WORDS chokeberry, saskatoon berry, antioxidant activity, anthocyanins, vitamin C, HPLC, UV/VIS/NIR, EPR spectroscopy 3 BUTOROVÁ, L. Charakterizace plod ů aronie ( Aronia melanocarpa ) a muchovník ů (Amelanchier alnifolia). Brno: Vysoké u čení technické v Brn ě, Fakulta chemická, 2014. 103 s. Vedoucí diplomové práce Ing. Eva Vítová, Ph.D. PROHLÁŠENÍ Prohlašuji, že jsem diplomovou práci vypracovala samostatn ě a že všechny použité literární zdroje jsem správn ě a úpln ě citovala. Diplomová práce je z hlediska obsahu majetkem Fakulty chemické VUT v Brn ě a m ůže být využita ke komer čním ú čel ům jen se souhlasem vedoucího diplomové práce a d ěkana FCH VUT. …………………………………………. podpis studenta Pod ěkování: Na tomto míst ě bych ráda pod ěkovala vedoucí diplomové práce Ing. Ev ě Vítové Ph.D. a konzultantce RnDr. Milen ě Vespalcové Ph.D. za odborné vedení ochotnost, čas a cenné rady. Dále bych cht ěla pod ěkovat Ing. Martinu Polovkovi PhD. a Ing. Blance Tobolkové z VÚP v Bratislav ě za vst řícnost, přátelský přístup, laboratorní zázemí a cenné rady p ři zpracovaní experimentální části práce. 4 OBSAH 1 ÚVOD ................................................................................................................................ 7 2 TEORETICKÁ ČÁST ..................................................................................................... 8 2.1 Aronia melanocarpa – Temnoplodec černoplodý ...................................................... 8 2.1.1 Chemické složení ............................................................................................... 9 2.1.2 Lé čivé ú činky ................................................................................................... 17 2.1.3 Nežádoucí ú činky ............................................................................................. 19 2.1.4 Použití ............................................................................................................... 19 2.2 Amelanchier alnifolia – Muchovník olšolistý .......................................................... 21 2.2.1 Chemické složení ............................................................................................. 22 2.2.2 Lé čivé ú činky ................................................................................................... 25 2.2.3 Nežádoucí ú činky ............................................................................................. 26 2.2.4 Použití ............................................................................................................... 26 2.3 Instrumentální techniky vhodné pro charakterizaci plod ů vybraného ovoce ........... 27 2.3.1 UV-VIS-NIR spektroskopie ............................................................................. 27 2.3.2 Vysokoú činná kapalinová chromatografie (HPLC) ......................................... 28 2.3.3 Elektronová paramagnetická rezonance (EPR) ................................................ 30 2.4 Metody vhodné pro charakterizaci plod ů vybraného ovoce .................................... 34 2.4.1 Stanovení celkového obsahu polyfenolických látek ........................................ 34 2.4.2 Stanovení celkových anthokyan ů ..................................................................... 35 2.4.3 Stanovení jednotlivých anthokyan ů ................................................................. 37 2.4.4 Stanovení barevných charakteristik v systému CIE L*a*b* ............................ 39 2.4.5 Stanovení koncentrace vitaminu C ................................................................... 41 2.4.6 Metody vhodné pro stanovení antioxida ční aktivity ........................................ 42 2.5 Statistické metody .................................................................................................... 47 3 CÍLE DIPLOMOVÉ PRÁCE ....................................................................................... 48 4 EXPERIMENTÁLNÍ ČÁST ......................................................................................... 48 4.1 Analyzované vzorky ................................................................................................. 48 4.2 Laboratorní vybavení ..............................................................................................
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