Primary Study On The Preparation And Safety Of High Concentration Garlic Oil Injection

Garlic is the underground bulbs of Allium sativum Linn.Garlic oil is the volatile oil extracted by steam distillation or organic solvent.Garlic oil has the function of antisepsis and anti-inflammation,cancer prevention and anticancer,cardiovascular disease prevention and treatment,protecting liver,increasing physical immunity,and anti-aging.At present,garlic preparations mainly include American garlic oil soft capsules,Japanese garlic ingot,German and Hungarian garlic oilβ-cyclodextrin inclusion complex,dehydrated garlic powder,garlic slices, garlicin microcapsule and garlicin injection.DATS is a single component by synthesized,and the injection preparation was from local standard to national standard in 2004 and 2005 respectively. Despite of the achievement of studying on garlic submicron emulsion injection,the research is far from production and application.Powder,pills,tablets,capsules are main in existing preparations,the route of administration is limited to a great extent since Garlic oil is slightly soluble in water.In order to get rapid and stable effects,Garlic oil is hoped to be prepared injection for intramuscular injection or intravenous injection.Now garlic oil injection is limited or withdrawn from markets for its potential hemolytic toxicity caused by Tween-80,a kind of solubilizing agent used to improve the solubility of Garlic oil. This article chose garlic oil by Inclusion of HP-β-CD to prepare injection in order to improve garlic oil concentration in garlic oil injection,improve the stability of Garlic oil,reduce irritations and toxicities and provide safe and effective higher concentration injection.The principal researches were as follow:Establishment of method for determination of garlic oil in high concentration garlic oil injection:By HPLC.The column used was Hypersil BDS-C18(200mm×0.46mm,5μm),with the mobile phase of acetonitrile-water(62:38)at the detection wavelength of 220nm.The column temperature was 35℃,and the flow rate was 1.2 ml/min.The good linearities were achieved over the concentration(1.4929～0.0149 mg/mL)for the sample.The precision,recovery and repeatability conforming to prescribed. Study on the preparation and preliminary evaluation the stability of high concentration garlic oil injection:Through orthogonal test,the preparation of high concentration garlic oil injection was selected.The grinding method and the best preparation technology was determined.The optimum conditions were the ratio of garlic oil and HP-β-CD of 1:10,grinding time of 1.5 h,preparation temperature of 30℃,pH of 8.5.Appearance characters of high concentration garlic oil injection was light yellow or yellow clearing liquid.The specification was 1 mL injection had 30mg garlic oil. Study on the stability of high concentration garlic oil injection:Initial equal-speed method was adopted to predict the period of validity.A reversed-phase HPLC method was developed for the determination of contents in injection at different temperature.It showed injection could be stored for 1.8 year at room temperature.It is suitable for clinical circumstances demand.The high concentration garlic oil injection is sensitive to photothermal.So it must store in suitable temperature and away from light.The accelerated test and long-term test showed the character,pH and content of high concentration garlic oil injection had no changes after six months.Primary study on the Safety of high concentration garlic oil injection:This paper determined LD₅₀by improved Karber method,accumulative toxicity test and tolerance test by progressive dose increase method.Bone marrow micronucleus test with 3 experimental groups at dosage of 181,73 or 36 mg/kg respectively and positive and negative control groups,spermatogenic malformation test with 3 experimental groups at dosage of 380,190 or 90 mg/kg/day respectively and positive and negative control groups.Hemolytic test by two percent red blood cell suspension.Observation of the irritation by pathological section after intramuscular injection the high concentration garlic oil injection.The results showed that LD₅₀of high concentration garlic oil injection on mice by i.m was 362.5mg/kg,and the confidence limit of 95%was 314.3～418.0 mg/kg.The cumulation coefficient was over 5.3.The other 3 results were all negative(P＞0.05).There was no hemolytic and no irritation.The high concentration garlic oil injection was safety and reliable,and it could be applied in muscle injection.Study on quality standard of high concentration garlic oil injection:Quality standard of high concentration garlic oil injection was established.After several rounds of review tests,the quality standard could effectively control the quality of the preparation,and laid the foundation for industrial production and clinical application.