田基黄总黄酮提取工艺优化、降血糖活性评价及成分分析OA北大核心CSTPCD
Optimization of extraction technology,evaluation of hypoglycemic activity and component analysis of total flavonoids from Hypericum japonicum
目的 优化田基黄总黄酮的超声波辅助提取-低共熔溶剂提取工艺,评价其体外降血糖活性,并初步分析其化学成分.方法 以氢键受体及供体组成、摩尔比、含水量为考察因素,以总黄酮得率为响应值,筛选田基黄总黄酮提取的最适低共熔溶剂;以液料比、超声功率、超声温度、超声时间为考察因素,以总黄酮得率为响应值,结合单因素实验和Box-Behnken响应面法优化田基黄总黄酮的提取工艺并验证;以阿卡波糖为阳性对照,测定田基黄总黄酮对α-淀粉酶、α-葡萄糖苷酶的体外抑制活性;采用超高效液相色谱法,通过对比对照品分析其化学成分.结果 最适低共熔溶剂为氯化胆碱-草酸(摩尔比1∶1,含水量30%);最优提取工艺为液料比52∶1(mL/g)、超声温度54℃、超声功率240 W、超声时间42 min,3次验证实验的总黄酮得率为(73.26±2.48)mg/g,与理论值(73.48 mg/g)的相对误差为-0.30%.田基黄总黄酮对α-淀粉酶和α-葡萄糖苷酶的半数抑制浓度分别为0.73、0.44 mg/mL,均高于阿卡波糖的0.23、0.15 mg/mL.田基黄总黄酮提取物中含有异槲皮苷、槲皮苷、槲皮素-7-O-α-L-鼠李糖苷、槲皮素.结论 优化所得田基黄总黄酮提取工艺稳定、可行,其提取物具有一定的体外降血糖活性,含有异槲皮苷、槲皮苷、槲皮素-7-O-α-L-鼠李糖苷等成分.
OBJECTIVE To optimize the ultrasound-assisted extraction-deep eutectic solvents technology of total flavonoids from Hypericum japonicum,evaluate its hypoglycemic activity in vitro,and analyze its chemical compositions preliminarily.METHODS The most suitable deep eutectic solvent for total flavonoids from H.japonicum was screened using the composition of hydrogen bond acceptor and donor,molar ratio,water content as factors,and the total flavonoid yield as the response value.Using liquid-solid ratio,ultrasonic power,ultrasonic temperature and ultrasonic time as factors,the yield of total flavonoids as response value,the extraction technology of total flavonoids from H.japonicum was optimized by single-factor experiments combined with Box-Behnken response surface method,and the optimum extraction technology was validated.Taking acarbose as the positive control,the inhibitory activities of total flavonoids from H.japonicum on α-amylase and α-glucosidase in vitro were determined.The chemical constituents of total flavonoids from H.japonicum were analyzed by UPLC combined with comparing the reference substances.RESULTS The most suitable deep eutectic solvent was choline chloride-oxalic acid(the molar ratio of 1∶1,the water content of 30%).The optimum extraction technology was as follows:the ratio of liquid-solid was 52∶1(mL/g),the ultrasonic temperature was 54℃,the ultrasonic power was 240 W,and the ultrasonic time was 42 min;the total extraction yield of total flavonoids from H.japonicum in 3 validation tests was(73.26±2.48)mg/g,the relative error of which with the theoretical value(73.48 mg/g)was-0.30%.The total flavonoids from H.japonicum could inhibit α-amylase and α-glucosidase with IC50 values of 0.73 and 0.44 mg/mL,respectively,which were higher than those of acarbose(0.23 and 0.15 mg/mL).UPLC analysis showed that the total flavonoids from H.japonicum contained isoquercetin,quercitrin,quercetin-7-O-α-L-rhamnoside and quercetin.CONCLUSIONS The optimized extraction technology of total flavonoids from H.japonicum is stable and feasible,and the extract has certain hypoglycemic activity in vitro and contains isoquercetin,quercitrin and quercetin-7-O-α-L-rhamnoside,etc.
周志红;胡勤;肖明明
中南大学湘雅医学院附属儿童医院(湖南省儿童医院)药学部,长沙 410007
中医学
田基黄总黄酮超声波辅助提取低共熔溶剂响应面法降血糖活性成分分析
total flavonoids from Hypericum japonicumultrasound-assisted extractiondeep eutectic solventsresponse surface methodhypoglycemic activitycomponent analysis
《中国药房》 2024 (016)
1972-1978 / 7
湖南省自然科学基金项目(No.2021JJ40279);湖南省卫生健康委科研立项课题(No.202113011086)
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