title: Modification and validation of a high-performance liquid chromatography method for quantification of Huperzine A in Huperzia crispata.
abstract: The present study describes a rapid and sensitive HPLC method for the quantification of huperzine A (HupA) in Huperzia crispata (Huperziaceae). The sample extraction and preparation involved a simple, time-saving, single-solvent extraction, with each sample being analyzed within 12 min. The mobile phase was ammonium acetate (0.1 M, pH 6.0)--methanol (64 + 36, v/v) at a flow rate of 1.0 mL/min. Detection was at 308 nm. The calibration curve was linear from 0.049 to 7.84 microg (R2 = 0.9997), with intraday and interday precision RSD of less than 2%. The extraction recovery rate was over 98.49%. Quantification of HupA was performed using this modified method, and the content of HupA was 1.86 times higher in the whole plant of H. crispata (218.17 +/- 1.55 microg/g) than in that of H. serrata (117.03 +/- 2.97 microg/g). In the whole plant of H. crispata, HupA mainly accumulated in the actively growing shoot tips, the apical bud, and the 10 youngest leaves, reaching 455.23 +/- 2.97 microg/g. The content of HupA in the samples from sunshine-sheltered sites was 3.45 times higher than in that from sunshine-abundant sites. The satisfactory results indicate that this modified method can be applied in the quality control of large-scale Huperziaceae plant extracts and that changes should be made in the cultivation of H. crispata so as to maximize the production of HupA.
authors.affiliation: Central South University, School of Biological Science and Technology, Molecular Biology Research Center, 110 Xiangya Rd, Changsha, Hunan 410078, People's Republic of China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Yu
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Shi
authors.forename: Jianan
authors.initials: J
authors.lastname: Huang
authors.forename: Yushun
authors.initials: Y
authors.lastname: Gong
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Liu
authors.forename: Weixin
authors.initials: W
authors.lastname: Hu
mesh_terms: D000470:Alkaloids; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D032501:Huperzia / Q000737:chemistry* / Q000254:growth & development; D010936:Plant Extracts / Q000032:analysis; D012717:Sesquiterpenes / Q000032:analysis*
chemical_list: D000470:Alkaloids; D010936:Plant Extracts; D012717:Sesquiterpenes; C050426:huperzine A