ABSTRACT
Objective To establish the fingerprint profiling of fruits of Macleaya cordata and study the method for its quality evaluation. Methods The fingerprint profiling of M. cordata fruits from different regions was established using high performance liquid chromatography (HPLC) based on the local standard which was the determination method of the content of protopine, allocryptopine, sanguinarine, and chelerythrine from Changsha of Hunan Province in 2009. The principal component analysis (PCA) and cluster analysis (CA) were performed to explore the correlation among the common fingerprint peaks, origins, and quality of M. cordata fruits. Results Eleven common fingerprint peaks were identified in the fingerprint profiling of chemical constituents of M. cordata fruits from different regions. M. cordata fruits produced from eight areas were classified into two classes by PCA and CA method, and there were five common peaks, including peak 5, 7, 8, 9, and 11 with significant contribution on the regional difference of the fingerprint. Also, common peak 6 was the right peak as the reference peak because of its less variation, appropriate retention time and intensity. Conclusion The fingerprint profiling of chemical constituents of M. cordata fruits established in this study has good precision, repeatability, and stability, which can be used to evaluate the quality of fruits of M. cordata from different producing areas.
ABSTRACT
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.
ABSTRACT
Objective: To study the antitumor molecular mechanism started on sanguinarine (San) and chelerythrine (Che) from Macleaya cordata. Methods: Determining the IC50 values of San and Che against A-549, HCT-8, and Bel-7402 cell lines by using MTT to clarify whether these two alkaloids are the active components in M. cordata; studying the interaction between human telomeric DNA and two alkaloids respectively by using UV-Vis, FL, and CD methods. Results: San and Che are the active antitumor components of M. cordata; San could induce HT4 to form antiparallel G-quadruplex completely with Ka of 5×108 and Che could induce HT4 to form antiparallel G-quadruplex partially with Ka of 930. Conclusion: One of the antitumor molecular mechanisms of M. cordata is that the two active components could induce human telomeric DNA to form G-quadruplex.