摘要
目的测定半枝莲标准汤剂中总黄酮的含量,评价其体外抗氧化活性,建立其指纹图谱,并进行化学模式识别分析。方法采用紫外-可见分光光度法测定半枝莲标准汤剂中总黄酮含量(以野黄芩素计);采用1,1-二苯基-2-三硝基苯肼(DPPH)自由基、2,2′-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)自由基清除实验考察半枝莲标准汤剂的体外抗氧化活性;采用高效液相色谱(HPLC)法,以野黄芩苷峰为参照峰,采用《中药色谱指纹图谱相似度评价系统(2004A版)》绘制16批半枝莲标准汤剂的指纹图谱并进行相似度评价,确定共有峰;采用SPSS 24.0软件进行Pearson相关性分析,筛选具有体外抗氧化活性的潜在物质,并以其为变量,采用SPSS 24.0、SIMCA 14.1软件进行聚类分析和主成分分析。结果总黄酮检测质量浓度的线性范围为2.106~21.06μg/mL(R;=0.9993);精密度、重复性、稳定性(120 min)试验的RSD均小于2%;加样回收率为100.62%(RSD=0.55%,n=6);总黄酮含量为0.634~1.053 mg/mL。DPPH自由基清除实验的半数抑制浓度(IC;)为1.120~3.602 mg/mL,ABTS自由基清除实验的IC;为0.684~1.327 mg/mL;相关性分析结果显示,半枝莲标准汤剂中总黄酮含量与DPPH自由基、ABTS自由基清除实验的IC;均呈负相关,相关系数分别为-0.976、-0.940(P<0.01)。16批半枝莲标准汤剂共有18个共有峰,相似度为0.964~0.997;共指认出其中4个共有峰,分别为野黄芩苷(8号峰)、野黄芩素(14号峰)、木犀草素(15号峰)、芹菜素(17号峰)。半枝莲标准汤剂HPLC指纹图谱中3~4、8~9、12~15、17号峰峰面积与DPPH自由基、ABTS自由基清除实验的IC;均呈显著负相关(P<0.05或P<0.01)。聚类分析结果显示,16批半枝莲标准汤剂可聚为两类,其中S2、S7~S8、S14~S16为一类,S1、S3~S6、S9~S13为一类;主成分分析将16批半枝莲标准汤剂分为两类,其中S2、S4、S7、S14~S16为一类,S1、S3、S5~S6、S8~S13为一类,S4、S13、S15样品的综合
OBJECTIVE To determine the contents of total flavonoids in Scutellaria barbata standard decoction,evaluate in vitro antioxidant activity,establish the fingerprint and conduct chemical pattern recognition analysis.METHODS The contents of total flavonoids in S.barbata standard decoction(calculated by scutellarein)were determined by ultraviet-visible spectrophotometry.In vitro antioxidant activity of S.barbata standard decoction was investigated by free radical scavenging tests of 1,1-diphenyl-2-trinitrophenylhydrazine(DPPH)and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)ammonium salt(ABTS);HPLC method was adopted.Using scutellarin as reference,the fingerprints of 16 batches of S.barbata standard decoction were drawn and evaluated by Similarity Evaluation System of TCM Chromatogram Fingerprint(2004 A edition),and the common peaks were determined;Pearson correlation analysis was carried out by using SPSS 24.0 software to screen substances with in vitro antioxidant activity.Taking them as variables,cluster analysis and principal component analysis were carried out by using SPSS 24.0 and SIMCA 14.1 software.RESULTS The linear range of total flavonoids were 2.106-21.06μg/mL(R;=0.9993);RSDs of precision,reproducibility and stability tests(120 min)were all lower than 2%;the recovery was 100.62%(RSD=0.55%,n=6);the contents of total flavonoids were 0.634-1.053 mg/mL.Median inhibitory concentration(IC;)of DPPH radical scavenging experiment ranged 1.120-3.602 mg/mL,and IC;of ABTS radical scavenging experiment ranged 0.684-1.327 mg/mL.The results of correlation analysis showed that the content of total flavonoids in S.barbata standard decoction was negatively correlated with the IC;of DPPH free radical and ABTS free radical scavenging experiment,and the correlation coefficients were-0.976 and-0.940 respectively(P<0.01).There were 18 common peaks in the fingerprints of 16 batches of S.barbata standard decoction;the similarities were 0.964-0.997.A total of 4 common peaks were identified,such as scutellarin(peak 8),s
作者
杜义龙
李赛
李艳荣
潘海峰
DU Yilong;LI Sai;LI Yanrong;PAN Haifeng(Hebei Key Laboratory of Traditional Chinese Medicine Research and Development,Chengde Medical University,Hebei Chengde 067000,China)
出处
《中国药房》
CAS
北大核心
2022年第4期425-432,共8页
China Pharmacy
基金
河北省高校省级重点学科建设项目(No.冀教高[2013]4号)
承德医学院自然科学研究计划项目(No.201824)。
关键词
半枝莲
标准汤剂
总黄酮
体外抗氧化作用
指纹图谱
主成分分析
聚类分析
Scutellaria barbata
standard decoction
total flavonoids
in vitro antioxidant effect
fingerprint
principal component analysis
cluster analysis
