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【目的】建立一种同时测定竹叶中13种黄酮类化合物(异荭草苷、荭草苷、异牡荆苷、牡荆苷、芹菜素、木犀草素、苜蓿素、7-甲氧基-苜蓿素、苜蓿素-7-O-葡萄糖苷、芹菜素-7-O-葡萄糖苷、Demethyltorosaflavone、芹菜素-7-O-葡萄糖-6″-O-鼠李糖苷、6-反式-(2″-O-α-鼠李糖基)乙烯基-5,7,3’,4’-四羟基黄酮)含量的高效液相色谱法,并用此方法对牡竹属10种竹种(麻竹、龙竹、牡竹、黄竹、梁山慈竹、花吊丝竹、云南龙竹、福贡龙竹、勃氏甜龙竹、版纳甜龙竹)竹叶黄酮类化合物进行分析比较,以期为黄酮类化合物的定性定量检测、牡竹属竹叶的开发利用及提取竹叶黄酮时的竹种选择提供参考。【方法】采用HPLC梯度洗脱分离黄酮类化合物,通过仪器精密度、LOD值、LOQ值、日内稳定性、日间稳定性及添加回收率对方法进行验证。竹叶样品用70%乙醇超声提取,经石油醚萃取后采用HPLC进行检测。【结果】各黄酮类化合物标准品分离效果良好,保留时间在13.23~54.67 min之间。13种黄酮类化合物标准品在0.01~500 mg·L-1的线性范围内呈良好的线性关系,相关系数R2在0.999 6~1.000 0之间。仪器精密度RSD在1.06%~2.55%之间,LOD值在0.01~0.10 mg·kg-1之间,LOQ值在0.03~0.34 mg·kg-1之间。方法准确性、日间与日内精密度良好,日内稳定性RSD在0.15%~0.67%之间,日间稳定性在0.44%~5.61%之间,13种黄酮类化合较稳定。7种黄酮类化合物的添加回收率高于70%,其余6种黄酮类化合物的添加回收率在39.76%~68.75%之间。采用此方法对牡竹属10种竹种竹叶黄酮类化合物进行分析,结果显示各竹种竹叶黄酮类化合物含量存在差异,除木犀草素、6-反式-(2″-O-α-鼠李糖基)乙烯基-5,7,3’,4’-四羟基黄酮和Demethyltorosaflavone未在部分竹种中检测到外,其余10种黄酮类化合物均在10种竹叶中检测到。13种黄酮类化合物总含量在419.94~5 155.84mg·kg-1之间,花吊丝竹叶中总黄酮含量最高,福贡龙竹含量最低。【结论】本文建立的同时测定竹叶中13种黄酮类化合物含量的高效液相色谱法简便、快速、准确,牡竹属10种竹种竹叶中黄酮类化合物含量较丰富,有利于开发利用。
【Objective】 To establish a method for simultaneous determination of 13 flavonoids (isoorientin, xyloside, isovitexin, vitexin, apigenin, luteolin, alfalfa, 7-methoxy - Alfalfa, Alfalfa-7-O-glucoside, apigenin-7-O-glucoside, Demethyltorosaflavone, apigenin-7-O-rhamnoside, 2 “-O-α-rhamnosyl) vinyl-5,7,3 ’, 4’-tetrahydroxyflavone) were determined by HPLC. Bamboo, bamboo, bamboo, yellow bamboo, Liangshan Ci bamboo, bamboo wire hanging flowers, Yunnan Longzhu, Fugong Longzhu, Boh Long sweet bamboo, Banana sweet bamboo) bamboo leaves flavonoids were compared, with a view to Qualitative and quantitative detection of flavonoids, bamboo development and utilization of bamboo leaves and bamboo leaves when flavonoids bamboo selection to provide a reference. 【Method】 Flavonoids were separated by HPLC gradient elution. The method was validated by instrument precision, LOD value, LOQ value, intra-day stability, daytime stability and added recovery rate. Bamboo leaves samples were extracted with 70% ethanol, extracted by petroleum ether and detected by HPLC. 【Result】 The results showed that the standard of flavonoids was separated well and the retention time was between 13.23 and 54.67 min. Thirteen flavonoid standards showed a good linearity in the linear range of 0.01-500 mg · L-1, with a correlation coefficient R2 of 0.999 6-1.000 0. The precision of the instrument was between 1.06% and 2.55% with the LOD values between 0.01 and 0.10 mg · kg-1 and the LOQ between 0.03 and 0.34 mg · kg-1. The accuracy of the method was good and the precision was good during the day and the day. The intra-day stability RSD ranged from 0.15% to 0.67%. The daytime stability ranged from 0.44% to 5.61%. Thirteen flavonoids were more stable. The recoveries of seven flavonoids were higher than 70%, and the recoveries of the other six flavonoids were between 39.76% and 68.75%. The results showed that the content of flavonoids in bamboo leaves varied with the exception of luteolin, 6-trans- (2 ”-O-α - rhamnosyl) vinyl-5,7,3 ’, 4’-tetrahydroxyflavone and Demethyltorosaflavone were not detected in some species of bamboo, the other 10 flavonoids were detected in 10 species of bamboo. The total contents of 13 flavonoids ranged from 419.94 to 5 155.84 mg · kg-1, and the content of total flavonoids was the highest among the four kinds of flowers, while the content of Fugong was the lowest. 【Conclusion】 Thirteen The content of flavonoids by high performance liquid chromatography is simple, rapid and accurate. The contents of flavonoids in 10 species of bamboo of Phyllostachys pubescens are rich, which is beneficial to exploitation and utilization.