Optimization Of Extraction By Response Surface Methodology And Effect Of Gigantol From Dendrobium Officinale On Human Gastric Carcinoma Cells

Gigantol is one of the important medicinal components of Dendrobium officinale Kimura et Migo.However,the content of gigantol in Dendrobium officinale is very low,so the explorations on the optimal extraction,accumulation rules,content,medicinal value,and other applications of gigantol in Dendrobium officinale have important theoretical and applied value.In this study,GC-MS was used to identify gigantol in Dendrobium officinale.HPLC-D AD was used to establish the detection method of gigantol and its methodology was verified.Then the method of ultrasound-assisted extraction of gigantol was performed and the contents of gigantol were measured in Dendrobium officinale and Dendrobium Fengdou from Huoshan area in Anhui Province.Moreover,the effects of extract in Dendrobium officinale and gigantol on proliferation of human gastric cancer cell line SGC-7901 were studied.It can provide a theoretical basis for the further development and utilization of Dendrobium officinale and its active ingredient gigantol.The main conclusions were as follows:1.Gigantol in Dendrobium officinale was identified by GC-MS.The control article of gigantol and Dendrobium officinale extract were separated at 38 min and had the same characteristic ions.These indicated that Dendrobium officinale contained gigantol.2.The HPLC-DAD method was established to detect gigantol in Dendrobium officinale.The detection conditions were as follows:mobile phase:chromatographic acetonitrile(B)and 0.1%formic acid aqueous solution(A);elution procedure:0 min～40 min,B from 5%to 80%;flow rate:1.0 mL/min;column temperature:25℃;injection volume:20 μL.The maximum ultraviolet absorption of gigantol was located at 279 nm.The verification results of the methodology showed that the detection method was reliable.3.The extraction process of gigantol in Dendrobium officinale was optimized,and the effects of methanol concentration,liquid ratio,temperature and time on the extraction efficiency were analysed.The optimum process conditions of the response surface optimization were as follows:the concentration of methanol was 92.98%,the ratio of material to liquid was 27.2 mL/g,and the extraction temperature was 41.41℃.4.The detection results of gigantol content in tissue culture seedlings of Dendrobium officinale showed that gigantol in the early development of Dendrobium officinale was the highest;and the detection results of gigantol content in cultivated seedlings of Dendrobium officinale showed that the content of gigantol in the leaves was significantly higher than that in the stems.The gigantol in the annual leaves was about 4.7942 μg/g,and the gigantol in triennial stems was 0.8978 μg/g.The comparison results of gigantol content in different varieties of Dendrobium Fengdou showed that the gigantol content of Dendrobium from Huoshan was significantly higher than that in the other three species.5.The treatment of gastric cancer cells with gigantol showed that the extracts of Dendrobium officinale and of four kinds of Dendrobium Fengdou could inhibit the human gastric cancer cell line SGC-7901 in different degrees except Dendrobium moniliforme.Gigantol showed an inhibitory effect on the human gastric cancer cell line SGC-7901,with IC50 values of 86.78 μg/mL(48 h)and 20.61 μg/mL(72 h),respectively;SGC-7901 cells were treated with a concentration gradient of gigantol and plate cloning experiments were performed.And the results showed that gigantol has an obvious inhibitory effect on the colony forming ability of SGC-7901 cells.