| Sanguinarine belongs to benzyl isoquinoline alkaloid, which is the main active ingredient of Macleaya. Sanguinarine has the effects of anti-bacterial, anti-inflammatory, anti-tumor, insecticide and strengthening the immune system, and so on. In the veterinary clinical it has mainly used to promote animal growth and prevent and treat diarrhea. This study aims to develop sanguinarine into a new type of nano preparation, to increase its solubility and improve the bioavailability, extend the time of its role in the body, thus reducing dosage and side effects.1. Preparation of sanguinarine nano powder:In this study, sanguinarine nanosuspension were prepared by ultrasonic method and high pressure homogeneous method. Through the excipients selection and process optimization, we determined the optimal preparation process as follows:take tween-801 mL (0.5%, w/v), add 180 mL of distilled water to dissolve them; take Sanguinarine bulk 4 g (2%, w/v) and add it to the distilled water contained stabilizers, mix, after high-speed probe at 8000 r/min for 5 minutes, and then, the suspension was homogenized by the high pressure homogenizer (200bar,500bar, 1000bar, 1500bar successively for 10 cycles); After that we added methyl cellulose (0.5%, w/v) solution 20mL to it, the sanguinarine nanosuspension were obtained after high-pressure homogenized times. Finally, place it in 60℃ vacuum drying 48h, to obtain the sanguinarine nano powder. Sanguinarine nanosuspension observed under transmission electron microscope, Sanguinarine is acicular and its size less than 1 micron. After sanguinarine nano powder by redispersion, we found that its shape and size did not change significantly compared to before curing, and no re-aggregation.2. Content determination of anguinarine nano powder:In this study, we established a HPLC method for determination the content of sanguinarine:Column:Kromasil C18 (150×4.6mm,5um); mobile phase:acetonitrile-0.1% phosphoric acid aqueous solution (28:72, V/V); flow rate:1mL/min; detection wavelength:284nm; column temperature:35℃; injection volume:10μL. The results prove that, in this condition, excipients and solvents were no interference to the content determination of sanguinarine. Sanguinarine have good linearity between 0.1 and 10.0 μg/mL, it also has good precision. It is simple and convenient to use the external standard method to determination of sanguinarine content. The results of the study show that this is a accurate and rapid method to determination of the sanguinarine content, which can be used for quality control of sanguinarine nano powder.3. Acute toxicity test of sanguinarine nano powder:In this study, up and down method was employed to carry out acute toxicity tests. The results show that LD50 of sanguinarine nano powder and sanguinarine bulk drugs for mice are 2471 mg/kg and 1643 mg/kg Sanguinarine content of sanguinarine nano powder and sanguinarine bulk drugs are 22.6% and 60%, so the LD50 convert into the actual content of sanguinarine are 558.45 mg/kg and 985.8 mg/kg. Compared to their actual content of sanguinarine shows that the acute toxicity has increased after sanguinarine bulk drugs prepared as sanguinarine nano powder, but all belong to low toxicity level, and the use of safety within the recommended clinical dose range.4. Subacute toxicity test of sanguinarine nano powders:Subacute toxicity test results show that there were no effects at each dose groups of sanguinarine nano powder on organ coefficients and hematological indexes in mice. Compared with the control group, there were no significant effect at low dose group (24.71 mg/kg) and middle dose group (49.42mg/kg) of Sanguinarine nano powder on biochemical indicators of mice. The high dose group (247.1 mg/kg) of the ALT and LDH were significantly increased compared with the control group. Therefore, we speculated that high dose of sanguinarine nano powder may have Slight damage to liver. However, histopathological examination of various organs were no obvious pathological changes. |
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