Department of Chemistry, University of Malakand, Chakdara, Dir Lower KPK, Pakistan.
BMC Biochem. 2018 Jun 26;19(1):5. doi: 10.1186/s12858-018-0095-7.
In this study Aesculus indica fruit was subjected to isolation of phytochemicals. Two antioxidants quercetin and Mandelic acid were isolated in pure state. The free radical scavenging and acetyl choline esterase inhibitory potential of the crude extract and sub fractions were also determined.
The antioxidant capacity of crude extract, fractions and isolated compounds were determined by DPPH and ABTS methods. Folin-Ciocalteu reagent method was used to estimate the total phenolic contents and were found to be 78.34 ± 0.96, 44.16 ± 1.05, 65.45 ± 1.29, 37.85 ± 1.44 and 50.23 ± 2.431 (mg/g of gallic acid) in crude extract, ethyl acetate, chloroform, n-hexane and aqueous fractions respectively. The flavonoid concentration in crude extract, ethyl acetate, chloroform, n-hexane and aqueous fraction were; 85.30 ± 1.20, 53.80 ± 1.07, 77.50 ± 1.12, 26.30 ± 1.35 and 37.78 ± 1.25 (mg/g of quercetin) respectively. The chloroform fraction was more potent against enzymes, acetyl choline esterase and butyryl choline esterase (IC = 85 and 160 μg/ml respectively). The phenolic compounds in the crude extract and fractions were determined using HPLC standard method. Chlorogenic acid, quercetin, phloroglucinol, rutin, mandelic acid and hydroxy benzoic acid were detected at retention times 6.005, 10.062, 22.623, 30.597, 35.490 and 36.211 in crude extract and different fractions. The ethyl acetate fraction was rich in the targeted compounds and was therefore subjected to column isolation. The HPLC chromatogram of isolated compounds showed single peak at specified retention times which confirms their isolation in pure state. The isolated compounds were then characterized by FTIR and NMR spectrophotometric techniques.
The Aesculus indica fruit extracts showed antioxidant and anticholine esterase inhibitory potentials. Two bioactive compounds were isolated in the pure form ethyl acetate fraction. From results it was concluded that the fruit of this plant could be used to minimize oxidative stress caused by reactive oxygen species.
本研究对印度七叶树果实进行了植物化学成分的分离。成功分离出两种抗氧化剂槲皮素和扁桃酸，且均为纯品。还测定了粗提取物及其各馏分的自由基清除能力和乙酰胆碱酯酶抑制潜力。
通过 DPPH 和 ABTS 法测定了粗提取物、馏分和分离化合物的抗氧化能力。福林-考尔法测定总酚含量，结果分别为 78.34±0.96、44.16±1.05、65.45±1.29、37.85±1.44 和 50.23±2.431（以没食子酸计，mg/g）。粗提取物、乙酸乙酯、氯仿、正己烷和水相部分的黄酮浓度分别为 85.30±1.20、53.80±1.07、77.50±1.12、26.30±1.35 和 37.78±1.25（以槲皮素计，mg/g）。氯仿馏分对酶（乙酰胆碱酯酶和丁酰胆碱酯酶）的抑制作用更强（IC=85 和 160μg/ml）。采用 HPLC 标准方法测定粗提取物和馏分中的酚类化合物。在粗提取物和不同馏分中检测到绿原酸、槲皮素、间苯三酚、芦丁、扁桃酸和羟基苯甲酸，保留时间分别为 6.005、10.062、22.623、30.597、35.490 和 36.211。乙酸乙酯馏分富含目标化合物，因此进行了柱分离。分离化合物的 HPLC 色谱图在指定保留时间处显示单个峰，证实其为纯品分离。然后用傅里叶变换红外光谱（FTIR）和核磁共振（NMR）分光光度技术对分离化合物进行了表征。
印度七叶树果实提取物具有抗氧化和抑制乙酰胆碱酯酶的潜力。从乙酸乙酯馏分中分离出两种生物活性化合物。结果表明，该植物的果实可用于减少活性氧引起的氧化应激。
