Researches On The Extraction, Isolation And Antibacterial, Anti-tumor Activity Of Curcumin

Turmeric, a traditional Chinese medicine, was used as the material in the experiment. We found the method to extract the effective constituents from turmeric and to identify the activity of the turmeric by exploring the measuring method, separation method and purification method through hyperpressure, the identification of volatile oil, the detection of bacteriostatic effect and antitumor activity of turmeric, and the determination of bacteriostatic effect of volatile oil.We detected the total content of curcumin through spectrophotomery and high performance liquid chromatography. We found that the content of curcumin was linear with the absorbance in spectrophotomery and peak area in the high performance liquid chromatography, and the experiment for the recovery rate and accuracy also showed that both methods were reliable, but after comparing the two results we found that the result of spectrophotomery was slightly higher than that of the high performance liquid chromatography.By using C₁₈ and C₃₀ column, we investigated the effect of seperation of three different curcumins. The results showed that by the use of methanol - water isocratic elution, the separation efficiency of C₃₀ column was far better than the C₁₈ columin in the same condition. We also got the baseline separation and the separation time was satisfying by using methanol- water as the mobile not using complex gradient elution procedure. According to the chemical structure, polarity, chromatographic retention time and the characteristic analysis of UV spectrum, the three peaks including dimethoxy curcumin, curcumin and turmeric to the methoxy factors were identified respectively.32 chromatographic peaks were separated from the volatile oil extracted from curcuma longa, and 22 peaks about 85.94% of the total which was tested were identified. The main components were ar-turmerone (16.06 %),β-curcumene (11.40%),β-phellandrene half times (7.27%), big root geranium ketone (6.86%), table-α-green celery-ene (5.85%), ar-onrcumene (5.43%),α-ginger flavonoids (5.38%), eucalyptol (5.25%) and benzyl alcohol (5.24%). The main composition of turmeric volatile oil was terpenoids, which is in consistence with the result of literatrues published already.Based on the determination of the effects of extraction solvent, solvent concentration, pressure, extraction time, solid-liquid ratio, extraction time on extraction ratio, an optimally combined model of hyperpressure extraction process was set up by orthogonal experiment method. The UHPE optimum process condition of extraction was determined: 70% ethanol, liquid to solid ratio of 45:1, the pressure is 350 MPa, extraction time 3.5 min. In the optimum extraction conditions, extraction was repeated 3 times, the total average extraction rate of curcumin 28.82 mg·g^-1, which was greater than the highest value (28.49 mg·g^-1) of the orthogonal experimental results. The results demonstrated that the UHPE optimum process condition by orthogonal experiment method was successful.The qualitative and quantitative determination to bacteriostasis of curcumin, extraction of curcumin, drilling method using plate and liquid antibacterial double dilution method were used. The results showed that certain concentration of curcumin had inhibiting effect on Escherichia coli, Bacillus subtilis, yeast, Aspergillus niger, Geotrichum. The effect of bacteriostatic action was E. coli> Aspergillus niger> Bacillus subtilis> Yeast > Geotrichum candidum. The minimum inhibitory concentration of E. coli was 0.78mg/ml, Aspergillus 1.56mg/ml, Bacillus subtilis 1.56mg/ml, yeast 3.125mg/ml, Geotrichum candidum 6.25mg/ml.The corresponding concentration of turmeric volatile oil extracted with petroleum ether was prepared, after petroleum ether was removed. The qualitative and quantitative determination to bacteriostasis was made by drilling method using plate and liquid antibacterial double dilution method. The results showed that turmeric volatile oil had inhibitory action on Escherichia coli, Bacillus subtilis, yeast, Candida Nguyen production and Geotrichum candidum. The minimum inhibitory concentration of E. coli, Bacillus subtilis, yeast, Candida Nguyen was 6.25mg/ml, but it had no inhibitory effect on Aspergillus niger and had the strongest inhibition on Geotrichum candidum.Human heptoma cell line HepG₂ were treated with different concentrations of curcumin, and the inhibiting effect of curcumin on HepG₂ cells proliferation was determined by MTT method. Results showed that curcumin could significantly inhibit the growth of hepatoma cell lines HepG₂, and performed a time and dose dependent relationship. The inhibiting effect increased with the increase of concentration in a certain range, but the effect would reduce after certain concentration. The inhibitory effect was best when the curcumin concentration was 20μmol/L and worked for 48 h.