Research On Preparation Of High Purity Crocetin Sodium And Its Bioavailability

Objective Crocetin sodium is a compound with potential sensitizing effect on anti-tumor therapy,which improves oxygen content in hypoxic tumor tissues,thereby enhancing the efficacy of radiotherapy and chemistry drugs.In this study,we investigated the preparation technology of crocetin salts and its bioavailability in vivo.This study could provide novel preparation technology of high purity crocetin sodium considered the condition of industrial production.And it would enhance the knowledge of crocetin-associated compounds on the therapy of malignant brain glioma.Methods])Process optimization and preparation of crocetin sodium:Comparison the advantages and disadvantages of using a variety of different processes such as water extraction and alcohol extraction,crude extraction by two commonly used macroporous resin,and LH-20 Sephadex or polyamide purification,finally,determine an effective,economical and suitable process for the production of high-purity crocetin sodium;2)Structure confirm of crocetin and crocetin sodium:The use of infrared absorption spectroscopy,nuclear magnetic resonance(1H NMR,13C NMR),and mass spectrometry to confirm the chemical structure of crocetin and crocetin sodium;3)Oral bioavailability study of crocetin sodium:Quarantine-eligible SD rats were divided into three groups crocetin sodium low,high-dose gavage group and intravenous group,take the plasma sample supernatant for in vivo pharmacokinetic analysis.Results 1)The final process route used in this study was water extraction.First,the herbs were dipped in water at room temperature for crude extraction.And the HZ-801 macroporous resin was used for purification.Then the purified water and 25%ethanol were used for elution,respectively.The crocus glycoside was collected in 70%ethanol.After then purified by 100 to 200 mesh polyamide column chromatography with the purified water and 20%ethanol.After freeze drying,the key intermediate crocus glycoside was received.Followed with hydrolyzing by 2%sodium hydroxide,the crocetin sodium was received.The purity reached 99.5%by HPLC detection.2)Spectrum results identified that the molecular formula of crocetin is C20H24O4,whose molecular weight was 328,showed 24 hydrogen proton and 20 carbon atom formants,respectively.Meanwhile,crocetin sodium is C20H22O4Na2,whose molecular weight was 372,with 24 hydrogen proton and 20 carbon atom formants.3)The measurement of crocetin sodium in the rat blood samples was established by HPLC.In HPLC detection,the endogenous impurities did not interference analysis.The linear range of crocetin sodium was 0.05-100 ?g/mL.The pharmacokinetic parameters of crocetin sodium were calculated by statistical moment analysis.Conclusion 1)The technology for the preparation of crocetin sodium in this study was economical and environmental protection.The equipments and reagents was common and the herbs used to isolate crocus glycoside-I were low-cost compared to the rare medicinal herb saffron crocus.And the purity of product was also reached high standards.2)The structure of product was confirmed by IR,1H NMR,1H-1H COSY,13C NMR,DEPT135,HSQC and mass spectrum.3)In this study,the bioavailability of 25 mg/kg and 100 mg/kg dose of crocetin sodium were 54.56%and 43.77%in Sprague-Dawley rats,respectively.