The Establishment of the Method of Cell Biochromatograpy and Analysis of the Active Ingredients from TongQiaoHuoXue Decoction Acting on the Neurocytes.

A novel strategy for screening active components in traditional Chinese medicines (TCM) using living cells and HPLC and GC analysis are proposed. The hypothesis is that when cells are incubated with the extract of Tongqiao Huoxue Decoction (TQHXD), a famous ancient prescription in TCM, the potential active components in the TQHXD should selectively combine with the cells, and the cell-combining components would be detectable in the extract of denatured cells. The identities of the cell-combining components could be determined by HPLC and GC analysis. Using the proposed approach, two characteristic active ingredients binding to the membrane of the PC12 cells are indicated. In the fingerprint of HPLC, there are two characteristic peaks. One active ingredient with its retention time was at around 70 min had been identified as muscone by HPLC, GC, which came from Moschus herb, the other active ingredient may come from the Allium fistulosum, its structure needs further research. Also, the protective effect of muscone on PC12 cells induced by Oxygen and glucose deprivation (OGD) had been explored. These results show that the pretreatment with muscone on PC12 cells observably increased cell viability, reduced the release of lactate dehydrogenase (LDH) and cell apoptosis. Combined with the pharmacodynamic study of muscone on neuroprotective effect, it could be identified as one of the effective components in TQHXD.

[Analysis of muscone from serum and cerebrospinal fluid of rabbits after intragastric administration of tongqiao huoxue granules].

OBJECTIVE: To analyse the muscone from serum and cerebrospinal fluid (CSF) of rabbits after intragastric administration of Tongqiao Huoxue granules (TQHXG) and provide the partial experimental reference on laying down quality standard of preparation of formula and offer new ideas on studying the relation between pharmacodynamics and its effective component of formula. METHODS: Muscone from TQHXG was qualitative identified by the methods of thin-layer chromatography (TLC) and gas chromatography (GC). Muscone from blood and CSF of rabbits was examined by GC after intragastric administration of TQHXG. Compared with standard reference material of muscone, blank serum and blank CSF, the segmental component absorbed into CSF of rabbits was confirmed. RESULTS: Spots of TQHXG on the chromatomap showed the same colour at the corresponding locality of the standard reference material. Detected by GC, chromatographic peaks appeared at the retention time of 4.8 minute in the chromatomaps of the serum with TQHXG, the CSF with TQHXG and standard reference material of muscone, while there were no chromatographic peaks in the chromatomaps of the blank serum, the blank CSF and methanol at the same time. The ingredient corresponding the chromatographic peaks at the retention time of 4.8 minute in the chromatomaps of the serum, CSF with TQHXG should be muscone. CONCLUSION: Muscone from TQHXG could penetrate blood brain barrier and could be one index of quality standard of preparation of TQHXG.