| In order to analyze the changes of compositions in young Leonurus heterophyllus (YLH) during growth and guide the harvest time, the dry whole plant of Leonurus japonicus Houtt and Leonurus liquid extract were studied in this research with modern analytical technology, which was to set up a quality control methodology in the fields of fingerprint and multi-component determination.Objective:(1) To set up one fingerprint method of Leonurus liquid extract (2) To set up one multi-component quantification analysis method of Leonurus liquid extract. (3) To set up one fingerprint method of Leonurus japonicus Houtt (L.japonicus) (4) To set up one multi-component determination of L.japonicus. (5) To research the dynamic regularity of chemical components in different growing periods of young Leonurus heterophyllus(YLH).Methods:(1) The chemical components in Leonurus liquid extract were isolate by the means of silica gel column chromatography, solvent extraction, etc. After that, methds of chromatography, spectroscopy and mass spectrometry were used to identify the structures of the components. Through teh optimized HPLC condition, the fingerprint of Leonurus liquid extract was set up and applied to analyze the Leonurus liquid extract (2) An HPLC method for the simultaneous determination of P-hydroxybenzoic acid, syringic acid, and hydrochloric acid leonurine was developed. (3) The polar compounds in L.japonicus were extracted and separated by the means of solvent extraction. (4) The determination of 3 components, P-hydroxybenzoic acid, syringic acid, and hydrochloric acid leonurine, was established. (5) the dynamic regularity of chemical components in different growing periods of young Leonurus heterophyllus(YLH) was compared.Results:(1) Because of the high contents in the water-saturated n-Buthyl extraction layer from Leonurus liquid extract,3 components were identified, which were P-hydroxybenzoic acid, syringic acid, and hydrochloric acid leonurine, respectively, through HPLC, spectroscopy and LC-MS methods. HPLC fingerprint of characteristic components of Leonurus liquid extract was established for the first time, Totally,6 common peaks were figured out, and 3 of them were identified. The results revealed that the sample fingerprints were similar with the standard fingerprint. (2) An HPLC method was set up for the simultaneous determination of P-hydroxybenzoic acid, syringic acid, and hydrochloric acid leonurine in Leonurus liquid extract. These 3 components showed good linearities in the ranges of 3.952-790.4ng (r=1.0000),10.427-2085.4ng (r=1.0000) and 6.983-1396.6ng (r=1.0000), with the average recoveries as 98.6%(RSD= 1.5%),97.8%(RSD=2.0%) and 97.9%(RSD= 1.8%), respectively. (3) The HPLC fingerprints of Leonurus japonicus Houtt were set up for the first time. Totally,6 common peaks were figured out, and 3 of them were identified, similar to the Leonurus liquid extract. The fingerprint showed that the correlations of Leonurus liquid extract and Leonurus japonicus Houtt were good. The results revealed that the sample fingerprints were similar with the standard fingerprint. (4) The quantitation method of P-hydroxybenzoic acid, syringic acid, and hydrochloric acid leonurine was set up in L.japonicus, The average recoveries of the 3 components were 99.0%(RSD= 1.5%),99.5%(RSD=0.7%),98.6%(RSD=1.3%), respectively. (5) The results of the contents determination in different growing periods showed that between 2.5th month to 3.5th month, a significant increase occurred in the contents of the three components, between 3.5th month to 4.5th month, a small decline occurred, and after the 5.5th month, rose again, then the contents gradually decreased. The results showed that the contents of the three active ingredients were all in high level in the 5.5th month after planting.Conclusions:(1) The HPLC characteristic fingerprint for the identification of Leonurus liquid extract was specific, simple and reproducible. (2) The established multi-component quantification analysis method could be used to conveniently and accurately control the quality of Leonurus liquid extract, and provide the basis for comprehensive evaluation and control of product quality. (3) There was good correlation between HPLC characteristic fingerprint of L.japonicus and Leonurus liquid extract,which was helpful to source the qualified herbs. (4) The multi-component quantification analysis method of Leonurus japonicus Houtt could used to select high-quality crude drug. (5) The dynamic changes of the 3 components in different growing periods of young Leonurus heterophyllus(YLH) could provide a guidance for the plantation and harvest of Leonurus japonicus Houtt. |
PDF Full Text Request