Technological Study On The Extraction Of Solanesol And The Evaluation Of The Anti-cancer Activity Of YIMIAOBAOERJINMI Capsules

Objective: 1. The technology of the extraction of solanesol from tobacco leaves was studied and a determination method of solanesol was established. 2. The anti-cancer activity and the acute toxicity of YIMIAOBAOERJINMI capsules was evaluated.Methods: 1. The content of solanesol was determined by means of HPLC method. The chromatographic column of DIKma C₁₈ (150 mm×4.6 mm, 5μm) was used. The mobile phase was methanol-iso-propanol (52: 48 ) . The flow rate was 0.8 mL/min and the detection wavelength was 210 nm. The injection volume was 20μL. The content was determined by the calibration curve. 2. The extraction procedure was studied with the dissociated solanesol as an indicator and factors such as solvents, method and time were investigated through orthogonal and comparison experiment. The saponification procedure was studied with the dissociated solanesol as an indicator and factors such as saponification substance, solvents, time, concentration, temperature, and the volume of saponification solution were investigated through orthogonal experiment. 3. The inhibition rate of YIMIAOBAOERJINMI capsules towards the in vitro growth of colon cancer cells (CT-26) and gastric cancer cells (MFC) of mice was determined by improved MTT method and median inhibition concentration IC50 was calculated. 40 mice of CT-26 colon cancer subcutaneous inoculation model and 40 mice of MFC gastric cancer subcutaneous inoculation model were randomly divided into four groups: negative control (taking equal volume of normal saline), positive control (taking Lentinan tablets, 0.5 mg/20 g/d; or Kanglixin capsules, 0.075 g/20 g/d), low dose group (taking powder of YIMIAOBAOERJINMI capsules, 0.0834g/20g/d) and high dose group (taking powder of YIMIAOBAOERJINMI capsules, 0.2501g/20g/d). Intragastric administration was made 5 days after the inoculation for continuous 8 days. The animals were executed 24 hours after the suspension of the drug. The tumors were striped and weighed to calculate the inhibition rate of tumor. The milts and thymus were taken out and weighed to calculate the milt index and the thymus index.Results: 1. It is indicated through methodical study that the linear range was 0.56～2.80μg for solanesol determination with a correlation coefficient r of 0.9995. The average recovery of assay was 100.2%(n=9). The precision gave a RSD of 1.19%. The content of dissociated solanesol in tobacco leaves extract was around 8%¹2%. 2. The recovery of solanesol varies much through different parameters of the extraction from the tobacco leaves and saponification procedures. The optimized extraction technology include: 10 times 95% ethanol, refluxing extraction at 50°C for 120 minutes or continuously ultrasonic extraction for 30 minutes. The optimized saponification technology include: saponification at 60°C for 4 hours in ethanol solution with 3 times 2.0 mol/L NaOH as saponification agent. 3. YIMIAOBAOERJINMI capsules demonstrate no obvious inhibition activity towards the in vitro growth of colon cancer cells (CT-26) and gastric cancer cells (MFC) of mice, while demonstrate obvious inhibition function towards the in vivo growth of colon cancer cells (CT-26) and gastric cancer cells (MFC) of mice. The high dose group shows a tumor inhibition rate of 30.26% towards colon cancer cells (CT-26) of mice, while the low dose group shows a rate of 25.74%. The high dose group shows a tumor inhibition rate of 29.35% towards gastric cancer cells (MFC) of mice, while the low dose group shows a rate of 20.53%, compared with that of the normal saline group.Conclusions: 1. The determination method of solanesol is feasible because of its good precision, accuracy and sensitivity. 2. YIMIAOBAOERJINMI capsules demonstrate obvious inhibition function towards the in vivo growth of colon cancer cells (CT-26) and gastric cancer cells (MFC) of mice, while no obvious dosage-effect relations are observed. 3. The ultrasonic extraction greatly improves the extraction rate from the raw materials. The selection of the technological parameters of the solanesol extraction from tobacco leaves and saponification procedures is very important and deserves further study.