不同形态乌药根的特征图谱比较及多成分含量测定

Characteristic Spectra and Multi-component Content Determination of Different Root Forms of Lindera Aggregate

关键词:  乌药  直根  块根  特征图谱  高效液相色谱法  含量测定

Key words:  Lindera aggregata (Sims) Kosterm.  taproot  root tuber  characteristic fingerprint  HPLC  content determination

摘要:

目的 建立不同形态乌药根指纹图谱和6种成分定量分析的测定方法，为其质量控制提供更可靠的方法和依据。方法 采用Agilent Zorbax SB-C₁₈色谱柱（4.6 mm×250 mm，5 μm），以乙腈-0.2%磷酸溶液为流动相，梯度洗脱，流速0.8 mL·min^-1；检测波长210 nm；柱温30℃；进样量10 μL。建立22批不同形态乌药根的HPLC指纹图谱，借助中药色谱指纹图谱相似度评价系统对指纹图谱进行分析，并对去甲异波尔定、乌药醚内酯、波尔定碱、异乌药内酯、钓樟内酯、山奈酚进行含量测定。结果 共标定了20个共有峰，22批样品相似度>0.980；去甲异波尔定、乌药醚内酯、波尔定碱、异乌药内酯、钓樟内酯、山奈酚在12.7~12 700（r=0.999 1），5.3~1 040（r=0.999 2），7.5~6 210（r=0.999 5），10.25~8 200（r=0.999 3），5.5~2 200（r=0.999 3），6.85~1 370（r=0.999 2）μg·mL^-1内与峰面积线性关系良好；含量测定结果表明，块根样品中去甲异波尔定、乌药醚内酯、波尔定碱含量较直根样品高，异乌药内酯含量较直根低，钓樟内酯和山奈酚含量无统计学意义上的差异。结论 该方法准确，稳定可靠，为乌药直根药用资源的开发和利用提供了科学依据。

Abstract:

OBJECTIVE To establish a method for the qualitative analysis of specific chromatogram and quantitative analysis of 6 components of different forms of Lindera aggregate, and to provide a reliable method and basis for the quality control of Lindera Aggregate. METHODS The determination was performed on Agilent Zorbax SB-C₁₈(4.6 mm×250 mm, 5 μm) column with mobile phase consisted of acetonitrile-0.2% phosphoric acid(gradient elution) at the flow rate of 0.8 mL·min^-1. The detection wavelength was set at 210 nm. The column temperature was set at 30℃, and the injection volume was 10 μL. The HPLC specific chromatogram were analyzed and the similarity was evaluated with "Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine". The contents of norisoboldine, linderalactone, boldine, isolinderalactone, linderane and kaempferol were determined. RESULTS Compared with the common pattern of fingerprints, the similarities of 22 batches of samples were>0.980 with 20 common peaks. The linear range was 12.7-12 700 μg·mL^-1(r=0.999 1) for norisoboldine, 5.3-1 040 μg·mL^-1(r=0.999 2) for linderalactone, 7.5-6 210 μg·mL^-1(r=0.999 5) for boldine, 10.25-8 200 μg·mL^-1(r=0.999 3) for isolinderalactone, 5.5-2 200 μg·mL^-1(r=0.999 3) for linderane, 6.85-1 370 μg·mL^-1(r=0.999 2) for kaempferol. The contents of norisobordine, linderane and boldine in root tuber of Lindera aggregate were higher than those in taproot of Lindera aggregate, while the contents of isolinderactone were lower in root tuber than taproot, there was no statistically significant difference in the contents of linderalactone and kaempferol. CONCLUSION The established method is accurate, stable and reliable. It can provide reference for a well-acceptable strategy for chemical comparison of taproot of Lindera aggregate.