Screening of active fractions from Curcuma Longa Radix isolated by HPLC and GC-MS for promotion of blood circulation and relief of pain

Ethnopharmacological relevance: Curcuma Longa Radix (Huangsiyujin in Chinese, HSYJ) has been used in clinical for thousands of years as a traditional Chinese medicine (TCM) and has the effects of invigorating the blood circulation, relieving pain, promoting qi circulation and relieving depression, besides, it is able to clear heart heat and cool blood and cure jaundice. The study aims to determine the active fractions of HSYJ for promotion of blood circulation and relief of pain, and to identify their chemical constituents. Material and methods: HSYJ was extracted by the systematic solvent method. The effects of promoting blood circulation and relieving pain were determined by measuring pain threshold (hot-plate test), latency of twisting and writhing times (acetic acid induced writhing test), and hemorheology of mice. Chemical constituents of the extractive fractions were analyzed by Gas Chromatography-Mass Spectrometer (GC-MS) or High Performance Liquid Chromatography (HPLC). Results: The results showed that five fractions (volatile oil fraction, petroleum ether fraction, ethyl acetate fraction, 1-butanol fraction and water fraction) all could increase pain threshold, prolong latency of twisting, decrease the writhing times and influence hemorheology. However, the effects of the volatile oil fraction and ethyl acetate fraction were better than other fractions. Three constituents, namely bisdesmethoxycurum, desmethoxycurumin and curcumin were identified from the active fractions by comparing with standard preparation through HPLC analysis. Forty-five compounds including aR-Turmerone, Curlone, Tumerone, Cyclohexene,1-methyl-4-(1-methylethylidene)-, trans-Sesquisabinene hydrate, (E)-Atlantone, alpha-curcumene and Zingiberene were identified from the volatile oil fraction by GC-MS. Conclusion: HSYJ possessed the effect of promoting blood circulation and relieving pain. Curcumins and their derivatives were the major constituents and might be the main bioactive ingredients for the promotion of blood circulation and relief of pain. The study provided evidence and formed a basis for the establishment of HSYJ comprehensive quality evaluation system related to traditional efficacy. It is beneficial to the quality control of HSYJ and its proprietary Chinese medicine.