摘要
利用沙棘粕为研究对象,经乙醇提取、大孔吸附树脂纯化获得沙棘粕醇提物(sea buckthorn seed extract,SBSE)。利用超高效液相色谱-串联四极杆飞行时间质谱(ultra-performance liquid chromatography coupled with quadruple time-of-flight tandem mass spectrometry,UPLC-QTOF-MS/MS)对SBSE进行定性研究,通过超高效液相色谱-串联三重四极杆质谱(ultra performance liquid chromatography coupled with triple quadruple mass spectrometry,UPLC-QQQ-MS)定量分析SBSE活性成分。最后研究分析SBSE对以D-半乳糖诱导的衰老小鼠的抗氧化功效,不同浓度SBSE灌胃衰老小鼠模型42 d后,比较小鼠肝脏组织中总抗氧化能力(total antioxidant capacity,T-AOC)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)活性以及丙二醛(malondialdehyde,MDA)和脂褐素(lipofuscin,LP)水平。结果表明:通过UPLC-QTOF-MS/MS分析方法共鉴定出24种化合物,其中与标准品比对共确认14种物质;利用UPLC-QQQ-MS法,采用负离子多反应监测方式,建立SBSE的14种化合物的定量分析方法,通过检测分析,SBSE中山柰酚及其衍生物所占比例最大,质量浓度为(667.94±5.61)μg/mL;其次为槲皮素及其衍生物((204.01±0.04)μg/mL),原花青素类物质为(33.97±0.49)μg/mL。经过42 d的衰老小鼠灌胃实验,与模型组相比,灌胃一定剂量SBSE可以提高小鼠肝脏组织中GSH-Px、SOD和CAT的活力;低剂量组小鼠肝脏组织中的T-AOC水平极显著高于模型组(P<0.01);3种灌胃剂量小鼠的肝脏组织LP和MDA含量均极显著低于模型组(P<0.01)。
This research studied the qualitative analysis of sea buckthorn seed extract (SBSE), prepared from sea buckthorn seed residue by ethanol extraction and macroporous resin purification, by ultra-performance liquid chromatography coupled with quadruple time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS). The bioactive components of SBSE were quantitatively determined by ultra performance liquid chromatography coupled with triple quadruple mass spectrometry (UPLC-QQQ-MS). Meanwhile, the antioxidant effect of SBSE in D-galactose-induced aging mice was studied. The mice were administrated by gavage with SBSE at different dosages for 42 days. Total antioxidant capacity (T-AOC), the activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) and the contents of malondialdehyde (MDA) and lipofuscin (LP) in liver tissue were measured. The results obtained were as follows: A total of 24 compounds were identified by UPLC-QTOF-MS/MS, 14 compounds of which were characterized by comparing with reference substances and quantitatively determined by UPLC-QQQ-MS in the negative ion multiple reaction monitoring (MRM) mode. Kaempferol and its derivatives were the most abundant constituents in SBSE ((667.94 ± 5.61) μg/mL), quercetin and its derivatives were in the middle ((204.01 ± 0.04) μg/m L), and procyanidins were the least abundant ((33.97 ± 0.49) μg/m L). The administration of SBSE to aging mice could increase the activities of GSH-Px, SOD and CAT in liver tissue. Compared with the D-galactose-induced model group, hepatic T-AOC activity in the low-dosage group was significantly increased (P < 0.01), and the levels of MDA and LP were decreased significantly by SBSE at three dosages (P < 0.01).
引文
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