A Systematic Research On Lentinan Injection Preparation Quality Standard

Lentinan is a kind of fungus polysaccharide. In1985, Lentinan was approved as a newanticancer drug,first in Japan,mainly used in the adjuvant therapy of cancer.people find thatLentinan has a good curative effect on gastric cancer,colon cancer,breast cancer,lungcancer therapy,can prolong the survival time of tumor patients;recent studies show thatlentinan has antiviral and anti-infection function.Lentinan production in China wasapprovaled for clinical application in1995.Lentinan injection preparation include injectionand sterile powder for injection.they have three different standards,WS3-149(Z-29)-95(Z),WS-1071（X-806）-2002and WS1（-X-152）-2004Z.The lentinan injection preparation fromthe same raw materials was performed in different standards,some implementationstandard for Chinese traditional medicine,some implementation standard for chemicaldrug,resulting in poor controllability.These standards have been implemented for manyyears, the check items are less and the determination methods are old.It is necessary toestablish a more efficient quality control standard,and ensure the safety,effective,stable,controllable of the lentinan injection preparation in clinical.According to the "Chinese Pharmacopoeia" and related literature,this paper made asystematic study on the lentinan injection preparation quality standard,and use thechromatographic technique as the main method.1.The improvement of determination method on current lentinan injection preparationquality standard.In the current quality standards, the determination method of lentinan (traditionalChinese Medicine) and lentinan for injection is titration, which operation is complicated,sensitivity and reproducibility is poor.As operation simple and good reproducibility,anthrone colorimetric method can be used for content determination of lentinan(traditionalChinese Medicine) and lentinan for injection.HPLC-RID is another effective method of thedetermination of lentinan injection preparation.it can avoid the interference from excipientsin lentinan injection preparation.But now it has no lentinan standard,the use of dextran as astandard to determinate the content will cause a certain error.2.Study on the projects outside the lentinan injection preparation quality standard.(1) Now the lentinan standards of injection preparation are lack of the check items onlentinan molecular weight.Studies show that the lentinan antitumor ability is closely related to Lentinan molecular weight. In this paper,we use the HPLC-RID for the lentinanmolecular weight determination of lentinan injection preparation.The results show that,different preparations of lentinan has different molecular weight.(2)The monosaccharide composition analysis is an important content of the study onpolysaccharide structure,activity and structure-activity relationship.This paper use theHPLC-RID method to study the monosaccharide composition of lentinan injectionpreparation.The results showed that,different preparations of lentinan has differentmonosaccharide composition.(3) The organic solvents such as chloroform,butanol,ethano were used in lentinanproduction process.According to the2010"Chinese Pharmacopoeia", the solvent residue ofchloroform,butanol,ethanol beyond the limits will be harmful to human body.This paperuse the GC to study the organic solvent residue limits control in lentinan injectionpreparation.The results show that,there is no residual solvent detected in the lentinaninjection preparation.(4)Benzyl alcohol is used in injection as an analgesic and a preservative,but if thecontent beyond the limits will be harmful to human body.Only lentinan injection(traditional Chinese Medicine) prescription contained benzyl alcohol,This paper use theHPLC–DAD for concentration determination of benzyl alcohol in lentinan injectionpreparation, The results show that,benzene methanol was only detected in the lentinan(traditional Chinese Medicine).(5)Lentinan is a polymerizing polysaccharide composed by glucose and other sugars.It is insoluble in water,and needs the solubilizer when lentinan injection isproducted.5-HMF is an aldehyde compounds,which hydrolysis from monosaccharidecompounds in high temperature or weak acidic conditions such as dehydration produced，and would damage the human striated muscle and visceral.Twain80is always used assolubilizer of insoluble drug for injection,but it have strong irritant, hemolysis andhypersensitivity.This paper use the HPLC-UV for the concent limit determination of5-HMF, Twain-80in lentinan injection preparation。The results show that,there are no5-HMF and Twain80detected in the lentinan injection preparation.(6)According to the2010"Chinese Pharmacopoeia"(appendix ⅨG) The Mooreosmotic concentration determination is inevitable items for injection preparation whenadded osmotic pressure regulator in prescription.This paper use the freezing point depression method for osmotic pressure molar concentration determination of lentinaninjection preparation.The results show that, different preparations of lentinan has differentMoore osmotic pressure.(7)Study shows that excessive intake of the heavy metal elements is harmful to humanbody,The heavy metals in injection preparation is usually originate from rawmaterials,auxiliary materials and production equipment. So it is necessary to limit thecontents of heavy metal elements in lentinan injection preparation.This paper use theICP-MS to determinate the metal elements in lentinan injection preparation.The resultsshow that, the contents of heavy metal elements in lentinan injection preparation did notexceed the standard.3.The influence of immunoregulation in rats after the lentinan injection preparationwas intraperitoneal injected.The mononuclear phagocytic system has a strong and rapid nonspecific phagocyticclearance ability of allogeneic particles,pathogens and tumor cells in the body.Studies showthat the immune function and molecular weight of lentinan are related.So thepharmacological activity of lentinan can be characterize by determinating the function ofmononuclear phagocyte system.This paper compare the strength of effect on mouse carbonparticle clearance, reflect the effect of lentinan injection on monocyte macrophage function,The results show that, lentinan for injection of880000Da of immunologic modification isstronger than the weight average molecular weight of320000Da lentinan injection(medicine and traditional Chinese Medicine).According to the above results,we developed the Systematic quality standard of thelentinan injection preparation,and it will be ensure the safety,effective,stable,controllable ofthe lentinan injection preparation.