博落回根中生物堿的组织化学定位研究

Translated title of the contribution: Histochemical localization study of alkaloids in root of Macleaya cordata

左姿, 郑亚杰, 梁之桃, 邹剑峰, 曾建国*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

目的:建立激光显微切割结合HPLC-MS/MS技术测定博落回根不同组织中血根堿、原阿片堿、别隐品堿、白屈菜红堿、二氢白屈菜红堿和二氢血根堿的定量分析方法,开展博落回根的组织化学研究。

方法:利用激光显微切割技术将两年生不同生长期博落回根的木栓层、皮层、韧皮部、木质部维管束和木射线进行切割,HPLC-MS/MS对切割的组织进行目标生物堿的检测。结果6种生物堿均有较宽的线性范围和良好的线性关系(r2>0.996 6);日内、日间精密度RSD均小于2.4%;检出限和定量限最高分别为0.29、0.57 ng/m L;重复性试验RSD均小于13.2%;加样回收率为85.1% 1 33.9%,RSD值为9.4% 2 0.7%。苗期根的上述5个组织中6种生物堿的总量分别为0.14、0.10、0.07、0.16、0.11μg/mm2;花期分别为0.38、0.22、0.15、0.41、0.26μg/mm2;果期分别为0.46、0.29、0.27、0.55、0.22μg/mm2;果后期分别为0.52、0.29、0.24、0.41、0.23μg/mm2。

结论:基于激光显微切割和LC-MS/MS的组织中生物堿的定量分析方法分辨率高、专属性强、灵敏度好、准确可靠;博落回根中6种生物堿主要分布在木栓层和木质部维管束。 

Objective: To develop the quantitative analysis method combined with laser microdissection and LC-MS/MS technique for determinating sanguinarine, protopin, allocryptopine, chelerythrine, dihydrochelerythrine, and dihydrosanguinarine in the roots of Macleaya cordata, and performe the histochemical study on M. cordata roots. 

Methods: The technology of laser-microdissection was used to dissect the cork, cortex, phloem, xylem vascular bundles, and xylem rays from 2-year-old M. cordata roots at different growth stages, the technique of LC-MS/MS was applied to detecting the contents of targeted alkaloids in the microdissected tissues. Results: The six kinds of alkaloids possesed a wide linear ranges and a good linear relationship (r2 > 0.996 6); The RSD of intra- and inter-day precision was all less than 2.4%; The highest limits of detection and quantification were respectively 0.29 and 0.57 ng/mL; The repeatability RSD was below 13.2%. The recovery varied from 85.1% to 133.9%, and its RSD ranged from 9.4% to 20.7%. The total amounts of six kinds of alkaloids in the above five tissues were respectively 0.14, 0.10, 0.07, 0.16 and 0.11 μg/mm2 for seedling stage root; 0.38, 0.22, 0.15, 0.41 and 0.26 μg/mm2 for flowering stage root; 0.46, 0.29, 0.27, 0.55 and 0.22 μg/mm2 for fruit stage root, and 0.52, 0.29, 0.24, 0.41 and 0.23 μg/mm2 for latering fruit stage root. 

Conclusion: The integrated method is high-resolution, specific, sensitive, and reliable for the quantitative analysis of alkaloid in tissues; The six kinds of alkaloids are mainly located in corks and xylem vascular bundles from the roots of M. cordata.

Translated title of the contributionHistochemical localization study of alkaloids in root of Macleaya cordata
Original languageChinese (Simplified)
Pages (from-to)1785-1790
Number of pages6
Journal中草药
Volume47
Issue number10
DOIs
Publication statusPublished - 28 May 2016

Scopus Subject Areas

  • Pharmaceutical Science
  • Drug Discovery
  • Complementary and alternative medicine
  • Pharmacology (medical)

User-Defined Keywords

  • Alkaloids
  • Histochemistry
  • HPLC-MS/MS
  • Laser-microdissection
  • Macleaya cordata (Willd.) R. Br.

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