Determination And Toxicity Of Naphthoquinones In Lithospermum Erythrorhizon

ObjectiveTo establish a HPLC method for the determination of Arnebia euchroma in three kinds of naphthoquinone compounds shikonin, acet ylshikonin and β,β-dimethylacry lshikonin content;To investigate the acute toxicity and cumulative toxicity of Arnebia euchroma naphthoquinone compounds effects in mice and discusses its main toxic target organs.MethodsDetermination of three kinds of naphthoquinone compounds shikonin, acetyl shikonin and β,β-dimethylacrylshikonin of the Arnebia euchroma content by RP-HPLC method.Acute toxicity:Using the modified Karber’s method for determination of LD50and LD5095%confidence limit, observed after administration for14days to observe and record the time of mice from death, death time and number.Cumulative toxicity:With the accumulation coefficient method to test and cal culate the accumulation coefficient, Observation and detection of normal physiolo gical and growth, viscera index, histopathology changes, serum biochemical indexes during the test in mice.ResultsThe linear response ranges of shikonin, acetylshikonin and β,β-dimethy lacryl shikoninwere19.4～194.0μg/ml(r=0.9992),26.4～264.0μ.g/ml(r=0.9993),19.0-190.0μg/ml(r=0.9998),respectively.The average recoveries were100.25%,101.07%,99.45%,respectively.RSDwerel.75%,1.96%,1.70%respectively.Arnebia euchroma of shikonin, acetylshikonin and β,β-dimethylacrylshikonin average content0.092%,1.217%,0.572%.The Arnebia euchroma naphthoquinone compounds single lavage LD50in mice=3.48g/kg and LD5095%confidence interval was from3.16to3.80g/kg.Cumulative toxicity test results showed that Arnebia euchroma naphthoquinone compounds in the mice inner accumulation coefficient K=3.76, moderate accumulative toxicity.The cumulative17days intragastric administration of Arnebia euchroma naphthoquinone compounds13.08g/kg can cause reduction food intake and weight loss in mice, the liver, spleen and gastrointestinal tissues viscera index compared with the control group there were significant differences, histopathological observation found that it had different degrees of injury and disease, female mice uterus index and ovarian index decreased, but the organization without the occurrence of pathological injury, Compared with the control group, the experimental group mice serum TP, ALB, A/G decreased significantly, ALT and AST were significantly increased.ConclusionsThis experiment is to establish a method for determination of Arnebia euchroma naphthoquinone compounds by high performance liquid chromatography inshikonin, acetylshikonin and β,β-dimethylacrylshikonin conten, this method has the advantages of simple operation, and the linearity, precision, repeatability is good. Study on toxicity of the extract was found, a single intragastric administration of LDso=3.48g/kg in mice, the accumulated gavage in mice inner accumulation coefficient K=3.76, moderate accumulative toxicity, its main target organ toxicity to liver, spleen and gast rointestinal tract. This provides reference for the identification of Arnebia euchroma select Pharmacopoeia other relatively stable HPLC components; it is not only for the future of Arnebia euchroma petroleum ether extract attenuated provides the basis, but also provided a guarantee for the rational development and utilization of Arnebia euchroma security; At the same time, It is provide technical support for showed that the composition of naphthoquinone in Arnebia euchroma, and screening of toxic components of this extract, and found the monomer compound properties of stability, development mechanism clear, high curative effect, low toxicity of Shikonin class compound new drug.