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| Gas chromatograph quantification and osteoblast cells stimulation study on lipophilic products from Bushen Huoxue Recipe |
| QIAN Yafang1，2 SHAN Letian3 JIN Hongting3 GU Mancang2 |
1.Preparation Center, Zhejiang Hospital of Traditional Chinese Medicine, Zhejiang Province, Hangzhou 310006, China;
2.College of Pharmacy, Zhejiang Chinese Medical University, Zhejiang Province, Hangzhou 310053, China;
3.Institute of Orthopaedics and Traumatology, Zhejiang Chinese Medical University, Zhejiang Province, Hangzhou 310053, China |
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Abstract Objective To establish determination method of lipophilic components in Bushen Huoxue Recipe by gas chromatography(GC), and investigate the activity of this component to induce osteoblast differentiation. Methods Multiply supercritical fluids were employed to extract lipophilic products SFEⅠ and SFEⅡ from Bushen Huoxue Recipe. Linoleic acid and oleic acid were determined by GC. The proliferation and ALP activity of osteoblast cells were explored after exposure to drug-loaded serums from SFEⅠ and SFEⅡ treated rats. Results Methyl linoleic acid and methyl oleic acid had good linear relationship within the inlet range of 0.2-1.2 μg. Methodological findings met the requirements. Linoleic acid and oleic acid content of SFEⅠ were 318.51-328.64 mg/g and 454.25-460.82 mg/g. Linoleic acid and oleic acid content of SFEⅡ were 205.22-211.32 mg/g and 228.55-235.13 mg/g. The drug-loaded serum from SFEⅠ and SFEⅡ treatment group could significantly stimulate the proliferation and ALP activity in MC3T3E1 cells in vitro compared with that of ethanol-water extraction groups (P < 0.05). Conclusion The supercritical fluid extracts of Bushen Huoxue Recipe SFEⅠ and SFEⅡ are mainly linoleic acid and oleic acid, and the osteoblast differentiation activity is better than that of alcohol extract.
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