Preparation Technics Of Anthraquinone Of Rhizoma Solid Lipid Nanoparticles And The Study Of Intestinal Absorption

Objective: Polygonum cuspidatum used as a traditional Chinese medicine, with the effects such as promoting blood circulation and relieve pain,clearing heat and dampness, releasing toxins and holding back coughing. With the development of modern medicine, the study of Polygonum cuspidatum has entered the era of the monomer composition studies. Modern pharmacological studies indicate: Resveratrol and Resveratrol glycosides from the stilbene ingredients of polygonum cuspidatum have the effects such as dilation of blood vessels, inhibit platelet formation, improve microcirculation of organism, hold back cough,etc.The anthraquinone substances polygonum cuspidatum have effect of anti-tumor, antibacterial, there are reports that anthraquinones in Polygonum cuspidatum inhibit the role of diabetic nephropathy. Because Anthraquinones in Polygonum cuspidatum has poor water solubility, in vivo absorption incomplete and other shortcomings, this experiment was prepared knotweed Polygonum cuspidatum extract purified total anthraquinone, combined the characteristics of SLN that this new nano drug delivery systems can improve the solubility of poorly soluble drugs,the slowly absorbed in vivo, bioavailability, Polygonum cuspidatum solid lipid nanoparticles were prepared. Its preparation process, quality standards, initial stability, release in vitro and intestinal absorption mechanism were studied and provide a scientific basis for action of drug formulations in vivo.Methods:(1) Extract process of Polygonum cuspidatum The content of emodin and physcion were detected by HPLC, and the total anthraquinone was detecetd by UV spectrophotometry. In knotweed content index, Using single factor and uniform design, central composite design experiments,with the index of total anthraquinone in knotweed,prefer the alcohol extraction process.(2) The process of Polygonum cuspidatum total anthraquinone content as an indicator of a resin model, eluent concentration, the maximum amount of sample selection.(3)Technology and quality standard of SLN preparation Use the single factor experiments and the star point design experiment with the indexs of particle size, encapsulation efficiency, drug loading to prepare the best prescription of the SLN. The outward appearance of SLN was observed by the TEM morphology, the particle size, and the distribution of the potential size were observed by the Nano particle size analyzer. TLC method was established to qualitative the emodin and physcion in the SLN; The content of emodin and physcion was determined by HPLC.Centrifuge tube with the microfiltration was used to determination the drug loading and encapsulation efficiency of SLN.(4) The study of preliminary stability and in vitro release properties of the SLN SLN changes at different temperatureschange in particle size 0-2 months, potential, encapsulation efficiency and drug loading method with effect factors determined SLN optimum storage conditions, the dialysis bag method SLN investigated in vitro release characteristics.(5) The intestinal absorption of the rat use the characteristics of rats intestinal condition similar to human physiological, the choice of the small intestine of rats in vivo perfusion model way to HPLC assay SLN intestinal perfusate emodin and emodin A content ether, more emodin, changes in the concentration of the perfusate physcion determine their rat duodenum, intestinal absorption rate constant(Ka) jejunum, ileum, colon and apparent distribution coefficient(Papp), after the inspection to join P-gP inhibitors verapamil hydrochloride, knotweed SLN total anthraquinone emodin, physcion intestinal absorption of Ka, Papp value change, and compared with the free inhibitor group, explore P-gP emodin and physcion impact on intestinal absorption.Results:Best result of alcohol(1) The extraction process of total anthraquinone: 10 times the amount of 70% ethanol with two times of extaction, 2h per time;(2) The preferred purification process: D101 resin selection, sample concentration(in crude drug meter) 0.1g / ml, pH = 10, the sample volume 3BV resin volume; eluent of 70% ethanol, the elution flow rate of 2BV / h.(3) The best preparation of SLN and the study of quality standard : high pressure homogenization method(heat), homogenizing pressure 100 MPa, five times the number of cycles; optimized formulation: stearic 0.160 g, total anthraquinone 0.136 g, the precent of P188 is 4.6%, the mass fraction of T-80 0.5%, lecithin 0.45 g, the total volume of prepared SLN is 50ml; prepared SLN looks like the clear solution with the half a reddish brownnearly yellow, TEM observation of spherical or spherical particle size of about 430 nm, Zeta potential-23.1mv, encapsulation efficiency was 87.1% drug loading 5%(average of emodin and physcion).(4) Preliminary stability of SLN and in vitro release SLN placed relatively stable at 4 ℃, size and potential, encapsulation efficiency and drug loading changed little under,with the enviroment of 25 ℃ in 30 days after placement gelatinous precipitates, the particle size, potential, encapsulation efficiency and drug loading have changed greatly. SLN in the dialysis solution containing 40% ethanol 48 h cumulative release rate of 73%, in vitro release behavior meet the first order distribution.(5) in rat intestinal absorption of polygonum SLN anthraquinone emodin and physcion are easily absorbed substances, mainly in the intestinal absorption site for colon, the emodin and physcion are P-gp substrates of SLN.Conclusion: Through the study of extraction and purification of total anthraquinone, and the preparation process of SLN, we discover the process for their preparation are scientific, stable and suitable for industrial production. Subject pioneered SLN preparation for Herbal Extracts and compared the extract and extract SLN in vivo intestinal absorption, provide a strong basis for in vivo pharmacokinetics in the future.