Indiarto Rossi, Pranoto Yudi, Santoso Umar
Pak J Biol Sci. 2019 Jan;22(1):34-44. doi: 10.3923/pjbs.2019.34.44.
The content of polyphenols in cacao beans can be modified during the processing of cacao. This study aimed to obtain the fraction of cacao bean extract polyphenols with the highest antioxidant activity and bioactive compounds profile of extracts and their fractions on cacao beans.
The cacao beans (fermented for 5 days and unfermented) were blanched (5 min; 95°C), followed with defatted, freeze-dried and extracted uses 80% ethanol solvent. The extract obtained was then fractionated using n-hexane, chloroform, ethyl acetate, n-butanol and aqueous. Extracts and fractions obtained are calculated for yield, total polyphenol uses Folin-ciocalteu reagent, total flavonoid uses AlCl3, antioxidant activity uses DPPH and FRAP methods, functional group uses fourier transform infrared spectroscopy (FTIR) and polyphenol compound profiles uses UHPLC-MS/MS.
The results showed that the aqueous fraction had the highest yield but lowest chemical content and antioxidant activity. The unfermented cacao beans extract undergoing fractionation using ethyl acetate showed polyphenol content, flavonoids, DPPH free radical scavenging activity and highest ferric reducing activity. The FTIR analysis showed that the cacao bean extract and its fractions had O-H, C-H, C=O, C=C and C-O-C functional groups. Cacao beans extracts and ethyl acetate fractions were dominated by procyanidin compounds, especially dimer B2. Cacao bean fermentation caused a decrease in procyanidin compounds (monomer to nonamer) and alkaloids (theobromine and caffeine).
Unfermented cacao bean extraction is then followed by fractionation with ethyl acetate solvent, obtained the fraction with highest chemical and antioxidant activity.
可可豆中的多酚含量在可可加工过程中会发生变化。本研究旨在获得具有最高抗氧化活性的可可豆提取物多酚馏分,以及提取物及其馏分对可可豆的生物活性化合物谱。
将可可豆(发酵5天和未发酵)进行烫漂(5分钟;95°C),随后脱脂、冷冻干燥并使用80%乙醇溶剂进行提取。然后将得到的提取物用正己烷、氯仿、乙酸乙酯、正丁醇和水进行分馏。对得到的提取物和馏分计算其产率,使用福林-酚试剂测定总多酚含量,使用氯化铝测定总黄酮含量,使用DPPH和FRAP方法测定抗氧化活性,使用傅里叶变换红外光谱(FTIR)测定官能团,使用超高效液相色谱-串联质谱(UHPLC-MS/MS)测定多酚化合物谱。
结果表明,水相馏分产率最高,但化学含量和抗氧化活性最低。未发酵可可豆提取物用乙酸乙酯进行分馏后,其多酚含量、黄酮类化合物、DPPH自由基清除活性和铁还原活性最高。FTIR分析表明,可可豆提取物及其馏分具有O-H、C-H、C=O、C=C和C-O-C官能团。可可豆提取物和乙酸乙酯馏分以原花青素化合物为主,尤其是二聚体B2。可可豆发酵导致原花青素化合物(单体至九聚体)和生物碱(可可碱和咖啡因)减少。
未发酵可可豆提取后用乙酸乙酯溶剂进行分馏,得到化学和抗氧化活性最高的馏分。