Application Study On The Modern Quality Control Of Flos Lonicerae From Shandong Province

Flos Lonicerae, the dry flower buds of Lonicera japonica Thunb., is common used for anti-virus, anti-inflammation, and antioxidant. And Lonicerae are famous traditional Chinese medicines and have a long history. Shandong province is one of main genuine producing areas in China, but the varieties used are in chaos. Several varieties are usually planted in one place or appeared in one batch of goods. The phenomenon is unfavorable for the quality control of Lonicerae. Therefore, the methods of quality assessment of Lonicerae from Shandong province were studied deeply and systematically, which provided evidences for the quality specification. The method should be helpful explore as a new quality control method for traditional Chinese medicine.Lonicerae were collected from different habitats, such as Shandong, Henan, Hebei, Hunan and Hubei. The Lonicera japonica Thunb. trueborn in Shandong Pingyi were considered as the main objects, so as the qualitative and quantitative analysis were studied.A reversed phase high performance liquid chromatography (RP-HPLC) method has been developed and validated for the quality evaluation of Lonicera japonica Thunb., Lonicera japonica var.chinensis, Flos lonicerae by fingerprinting analysis. Separations were achieved using a 250mm×4.6mm,5τm, reversed-phase column. The elution system consisted of two components:water (0.1% phosphoric acid) and acetonitrile with gradient elute. Column temperature was kept at 25℃. Analysis time was 60min. Blank tests and samples were detected at 238 nm as well as 190-400nm to gain the three dimensional pictures with the DAD. The results revealed that there were 13 common chromatographic peaks in Lonicerae. In this study, Similarity identification and Cluster analysis were studied based on the developed HPLC fingerprints. With the method of similarity identification, there was some differentia in Lonicerae between different territories and different varieties. And the result for similarity identification was coincident with the result of cluster analysis. The developed method proved practicable and reliable for quality control.The volatile oil from Lonicera japonica Thunb., Lonicera japonica var.chinensis and Flos lonicerae were studied by Headspace-Gas chromatogram- mass spectrum (HS-GC-MS) technique for the first time. Influences of headspace heating temperature, headspace heating time and sample weight on relative peak area of volatile compounds were investigated, and the HS-GC-MS method were developed and validated. There were 59 species of compounds in the Lonicerae with the established method. The constituents and contents had much difference for Lonicera japonica var.chinensis and other Lonicerae. HS-GC-MS can be used for the analysis of Lonicera japonica Thunb. With high precision, less sample consumption, no organic solvent extraction and highly automatic analysis process, the method was available for the analysis of Flos lonicerae.A HPLC method was developed for simultaneous determination of 4 constituents such as Chlorogenic acid, Sweroside, Hyperoside and Lutedin-7-O-glucoside in Lonicerae, collected from different habitats and different batches. The linear calibration curves were plotted with satisfied relative coefficients (r), which were 0.9994,0.9999,0.9975 and 0.9998. The average recoveries were in the range of 99.66-101.00% with RSD% less than 2.55%. Lonicerae were determined with the developed quantitative method. Following results were achieved from the analysis. The contents of Chlorogenic acid for all samples were in accordance with the rule of ChP., except one batch. The established assay method is simple, rapid, and reproducible and provides a quantitative basis for the quality assessment for Lonicerae. HPLC with on-line diode-array detector (HPLC-DAD) and electro spray ionization tandem mass spectrometry (HPLC-ESI-MS) was developed for simultaneous analysis of compounds in the Lonicerae. Combining the separation by HPLC, the characteristic of UV spectra, and the information of molecular structure provided by ESI-MS, eleven peaks were identified as Loganin, Swertiamarin, Chlorogenic acid, Sweroside, n-Caffeoylquinic acid, Secoxyloganin, Epi-vogeloside, Hyperoside, Lutedin-7-O-glucoside, Aldosecologanin and 4,5-dicaffeoylquinic acid. Study with correlation between HPLC-FP and GC-MS-FP for qualitative indentify, combined the cluster analysis for chemical pattern recognition to enhance the quality control system.The developed method with the modern spectrum technology, proved practicable and reliable for quality control of Lonicerae, which provided the qualitative and quantitative basis for the quality assessment. The method should be helpful explore as a new quality control method for traditional Chinese medicine (TCM).