The Research About HPLC Fingerprint Of Callicarpa Nudiflora In Hainan Province

ObjectiveIn order to achieve the Callicarpa nudiflora quality control, ensure the safety and the clinical application of slices and the related preparation effective, make a solid foundation to the further development and The production of sustainability, We carry out the research about Callicarpa nudiflora’s quality standards, it is including:literature reseach, HPLC Fingerprint, et al. Thus improve the quality evaluation standard of Callicarpa nudiflora, to establish a GAP planting base to provide scientific data to support, keep pace with the requirements of our Chinese herbal medicine quality standard today.MethodsIn this paper, we choose a high performance liquid chromatography (HPLC) fingerprint method, which is a fast, reliable and mature technology, to build a fingerprint of Callicarpa nudiflora. The chromatographic condition:mobile phase, detection wavelength, gradient elution program and testing time; The preparation methods of test sample solution:extraction method, the specises of extraction solvent, the dosage of extraction solvent, extracting time, the baseline of chromatogram, degree of separation of the chromatographic peaks, the number of the chromatographic peaks, et al. to be the evaluation indicators. We make the method of combining the relevant literature, retention time, the purty of peak and UV absorption to primary definite the compostion peak of Callicarpa nudiflora.10batches of Callicarpa nudiflora which comes from Hainan province are analyzed and evaluated by "The traditional Chinese medicine (TCM) chromatographic fingerprint similarity evaluation system2004A (research)" software, then common pattern generation is established.11batches Callicarpa nudiflora which were harvested in different times of Baisha county (3batches collected in2010are wild) do compare with the common pattern generation respectively. To the fingerprint patterns generations of other three areas of hainan province and hainan province common pattern generation, batches in different seasons, the distinction we can understand by the comparison.Callicarpa kwangtungsis, Callicarpa macrophylla, Callicarpa formosana and Callicarpa kochiana, which are also from Verbenaceae Callicarpa, are determinated by following the ensure method. Our objective is reflecting the specificity of Callicarpa nudiflora.ResultsThe HPLC Fingerprint conditions of Callicarpa nudiflora:the chromatographic column:Agilent TC-C18(250×4.6mm,5μm);mobile phase: acetonitrile:0.30%acetic acid; the flow rate:1.0mL/min; gradient elution; column temperature:30℃; the sample injection volume:20uL; detection wavelength:254nm. The preparation methods of test sample so]ution:2g Callicarpa nudiflora powder is weighed,50%methanol50mL, reflux extraction about30min in a water bath pot, which keep a temperature about80℃, wait until it is cooling, we filter it by a0.45u m filtration membrane, then the test sample is made.On the HPLC fingerprint of Callicarpa nudiflora, we select19peaks to be the common pesks and establish a common pattern generation. The HPLC fingerprint of samples have a high similarity with each other and the reference fingerprint. The result of the similarity calculation and system clustering analysis shows that there are differences between the samples from the same place(Baisha city) but not the same harvest time, samples of wild and cultivated have differences; give a priority to choose Lingao county, Haikou city, Wuzhishan city and establish GAP planting base; samples recoveried in different seasons, and that individual difference is bigger, the harvest time can be in the hot season (March-August).For the result of the study, we preliminary sure about that, the11th peak is the location of Acteoside(36.17min), the16th peak is Luteoline(64.42min), and the18th peak is5,4’-dihydroxy3,7,3’-trimethoxyl of flavone (92.37min). The chromatogram of Callicarpa nudiflora is unique when contrast with Callicarpa kwangtungsis, Callicarpa macrophylla, Callicarpa formosana and Callicarpa kochiana. It can achieve on the purpose of Callicarpa nudiflora authenticity identificantion.ConclusionIn this issue, we establish the HPLC fingerprint of Callicarpa nudiflora. this method can be used to distinguish between Callicarpa nudiflora and other Verbenaceae Callicarpa commonly used in the market, to the purple of reflecting Callicarpa nudiflora species specificity. For Callicarpa nudiflora in different regions, different harvest time, We can understand the internal difference by similarity clustering analysis and system evaluation. In combination with the content determination results, providing a scientific basis to the cultivation and raw material selection of Callicarpa nudiflora.