摘要
目的 建立诺日布-7散的高效液相色谱(High performance liquid chromatography,HPLC)指纹图谱和四种特征成分的含量测定方法。方法 采用HPLC法对10批次诺日布-7散建立指纹图谱,色谱条件为Agilent ZORBAX SB-C_(18)色谱柱(250 mm×4.6 mm,5μm),0.1%磷酸水溶液为水相,甲醇∶乙腈(1∶1,V/V)为有机相,梯度洗脱;检测波长为225 nm;流速为1 mL/min;柱温为30℃;进样量为5μL。采用该色谱条件对10批次诺日布-7散进行分析,标定共有峰并进行指认归属,并对没食子酸、栀子苷、异土木香内酯、土木香内酯4个特征成分进行含量测定。采用SPSS 25.0和SIMCA 18.0软件进行聚类分析(Cluster analysis,CA)、主成分分析(Principal components analysis,PCA)和偏最小二乘法-判别分析(Orthogonal partial least squares-discriminant analysis,OPLS-DA),对制剂的整体质量进行评价。结果 10批次诺日布-7散样品中共标记20个共有峰,指认6个共有峰,分别为没食子酸、栀子苷、柯里拉京、鞣花酸、异土木香内酯、土木香内酯,相似度在0.981~0.996之间;CA分析结果显示,10批次诺日布-7散样品分为3类:S1单独聚为一类;S2、S4、S6和S7聚为一类;S3、S5、S8、S9和S10聚为一类,PCA得到3个主成分的累积方差贡献率为86.553%,其得分图显示三类样品空间分布模式与CA完全一致。OPLS-DA模型进一步验证该分类,得分图中三类样品完全分离,且成功筛选出VIP>1的7个差异标志物峰。线性试验结果表明,没食子酸、栀子苷、异土木香内酯和土木香内酯的质量浓度与色谱峰面积线性关系良好;精密度、稳定性、重复性、加样回收率均符合方法学要求。10批次制剂中没食子酸、栀子苷、异土木香内酯、土木香内酯的含量依次为0.549±0.041、8.194±0.804、1.980±0.164、1.013±0.106 mg/g。结论 本研究建立的诺日布-7散含量测定和指纹图谱方法操作简便、结果可靠,为诺日布-7散质量标准提高提供数据支撑。
Objective To establish the high performance liquid chromatography(HPLC)fingerprint and a content determination method for four characteristic components in Noribu-7 Powder.Methods HPLC fingerprints were developed for 10batches of Noribu-7 Powder using an Agilent ZORBAX SB-C18 column(250 mm×4.6 mm,5μm)with 0.1%phosphoric acid aqueous solution and methanol:acetonitrile(1∶1,V/V)as mobile phases under gradient elution.Detection wavelength:225 nm;flow rate:1 mL/min;column temperature:30℃;injection volume:5μL.Common peaks were calibrated and identified,and gallic acid,geniposide,isoalantolactone,alantolactone 4 characteristic components were quantified.Cluster analysis(CA),principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were performed using SPSS 25.0 and SIMCA 18.0 software.Results 20 common peaks were marked in 10 batches,with 6 identified as gallic acid,geniposide,corilagin,ellagic acid,isoalantolactone and alantolactone.Similarities ranged between 0.981~0.996.The CA results showed that the 10 batches of Noribu-7 Powder were divided into 3 categories:S1 clustered alone;S2,S4,S6 and S7 clustered into 1 category;S3,S5,S8,S9 and S10 clustered into another category.PCA yielded 3 principal components with a cumulative variance contribution rate of 86.553%,and its score plot demonstrated that the spatial distribution patterns of the 3 sample categories were completely consistent with CA.The OPLS-DA model further validated this classification,with complete separation of the 3 sample categories in the score plot,and successfully screened 7 differential marker peaks with VIP>1.Linear relationships for gallic acid,geniposide,isoalantolactone and alantolactone were satisfactory.Precision,stability,repeatability and recovery met methodological requirements.The contents in 10 batches were determined as follows:gallic acid 0.549±0.041 mg/g,geniposide 8.194±0.804 mg/g,isoalantolactone 1.980±0.164 mg/g and alantolactone 1.013±0.106 mg/g.Conclusion The established HPLC fingerprint and content determination method are simple and reliable,providing data support for quality standard enhancement of Noribu-7 Powder.
作者
张梓月
朱雯钰
张瑶
马桂芝
ZHANG Ziyue;ZHU Wenyu;ZHANG Yao;MA Guizhi(College of Pharmacy,Xinjiang Medical University,Urumqi 830017,China;Department of Pharmaceutical Analysis,Xinjiang Medical University,Urumqi 830017,China)
出处
《新疆医科大学学报》
2025年第8期1137-1145,共9页
Journal of Xinjiang Medical University
基金
新疆维吾尔自治区重点研发计划项目(2017B03013-2)
新疆医科大学大学生自治区级创新训练计划项目(S20211076010)。
关键词
诺日布-7散
高效液相色谱
含量测定
特征成分
指纹图谱
Noribu-7 Powder
high performance liquid chromatography(HPLC)
content determination
characteristic components
fingerprint
