Molecular Biological Identification And Chemical Constituents Analysis Of Thirteen Chinese Medicinal Plants

Piper is the largest genus in the Pepper Family and the plants of this Genus are widely distributed throughout the tropical and subtropical regions of the world. Plants belonging to the Genus Piper are used medicinally in various manners, and are reputed in the Indian Ayurvedic system of medicine for their medicinal properties and in folklore medicine of Latin America and West Indies. In traditional folk medicine, many species of this genus are used for a variety of purposes including invigorating the circulation, reducing stasis, detumescence and as analgesics. To date, the majority of plants species found in this genus are used in the treatment of pain and central nervous system (CNS) disorders which has become a high incidence of disease. However, it’s difficult to identify plants in this genus precisely based on the subtle morphological characteristics, so new ways and discriminate characteristics to support the accuracy of source are urgently needed. Therefore, DNA sequence analysis and HPLC fingerprint technology are performed to identify the Piper species. Chemical constitutes in Piper species are analyzed in this study, too.In order to find DNA molecular markers for identification, we studied the rDNA ITS sequences variation patterns of the medicinal plants of Genus Piper in China. The results thereof showed that the homologous alignment of ITS sequences was lower than 66%between Genus Piper and outgroups. While within Genus Piper, it was higher than 81%. The identification standard is above 97%. Thus ITS sequence is applicable and reliable for identification of Piper at molecular level.A number of phytochemicals with anti-depression activities have been extracted from Piper plants by our group. A HPLC method for simultaneous determination of 10 active components in 12 Chinese Piper species was established. The method was precise, sensitive and showed a high accuracy for the quantification. In this study, it is the first reported occurrence of laetispicine in P. austrosinense, P. hancei, P. wallichii, and P. hongkongense. Meanwhile, the HPLC fingerprint of Piper was established, the common characteristics of the fingerprint of this genus and characteristics of 8 major species’fingerprint are applicable for further identification of Piper at chemical level.This study developed the methods of identifying Piper species both at molecular and chemical level, which could not only be useful for researching resources of bioactive compounds, but also provide a method to control TCM’s quality.