Preparetion Of Hypericin Injection And Its Quality Standards

Hypericin was first separated by Dietrich in 1891. S.Czerny named hypericin in 1911. Its chemical formula was determined in 1951 as 4, 4', 5, 5', 7, 7 '- six hydroxyl - 2, 2' - dimethyl - meso - naphtalin antithetical dianthrone. The molecular formula is C30H16O8 and the molecular weight is 504.43. Hypericin is blue black acicula and can decompose easily when heated up. It does not have fixed melting point.It must preserved without light because of its sensitivity to light. Hypericin is easy to dissolve in some organic solvent, such as pyridine, methyl alcohol, ethyl alcohol, ethyl acetate and appears orange with red fluorescence. Hypericin can be dissolved in the alkaline water and appear red when pH is below 11.5. When pH was above 11.5, It is green with red fluorescence. The functions of hypericin has been proved, such as treating depression, antibacterial, easing pain,hemostasia and so on. It is also a kind of endogenous photosencitizer and was adopted for remedying and diagnosing tumour. In addition, hypericin was confirmed that has strong activity on restraining RNA virus. So we can infer that hypericin will become development prospects natural medicine.A protein-bound complex consisted mainly of hypericin was developed successfully by Prof. Liang. It has been demonstrated that this complex can kill avian influenza virus (AIV) in vitro and in vivo. Preparation and establishment of quality standards of hypericin Injection was studied to take advantage of its pharmacologic function effectively and quickly.Hypericin Injection was prepared in compound solvent and its content was determined by HPLC. The Chromsphere C18 (4.6×150mm, 5μm) column was used to determine the content of hypericin. The mobile phase was methanol-acetonitrile-1% phosphate buffer (170:10:10).The flow rate is 1ml/min and the detection wavelength is 588nm. The calibration curve ranging from 0.04μg to 0.2μg was linear,The content of hypericin was measured to be 0.21% (equivalent to 1ml injection containing 8.97μg hypericin)and RSD was 1.07%. HPLC is sensitive, accurate, highly reproductive, and it is suitable for ananlysis of hypericin injection with the characteristic of sensitivity accuracy and repeatability.The quality standards test was carried out subsequently. The hypericin in injection were identified by TLC method. The contents of hypericin in injection were determined by HPLC. Practical experiences prove that hypericin in injection could be detected obviously by TLC and HPLC. TLC is a simple and accurate method and HPLC is a accurate and reliable method. The quality standard can be used for quality control of this product.Above the results, we can draw a conclusion that clear and transparent hypericin injection can be successfully prepared using compound solvent method. Thin-layer chromatography (TLC) and High performance liquid chromatography method (HPLC) were used to evaluate the quality standards of hypericin injection. This injection conforms to the stipulation in the appendix of the 2005 edition of Chinese Pharmacopoeia. This test is a foundation for the clinical use of hypericin injection,It will supply sientific basis and detection standards for the development of hypericin injection.