Toxicological Safety Evaluation Of Bound Polyphenols Of Seabuckthorn Berries

Seabuckthorn,also known as vinegar willow,is a wild shrub or small tree of the family Elaeagnaceae,genus Hippophae.It is reported that the polyphenols in seabuckthorn have significant effects including relieving angina pectoris,anti-tumor,anti-oxidation and anti-inflammatory.However,its systematical toxicological statistics are very scarce.Followed by the Procedure and Methods of Food Safety Toxicological Assessment GB-15193(China),the acute oral toxicity test,three genetic toxicity tests and repeated dose 28-day oral toxicity test were conducted to explore safe oral doses.Acute oral toxicity testAccording to Horn's method,forty ICR mice(half male and female)were randomly allocated to four groups treated with 2.15,4.64,10.00 and 21.5 g/kg.bw of Bound Polyphenols of Seabuckthorn Berries(SBBP)by oral gavage.The LD50 of SBBP for male and female mice is 5.84 g/kg.bw(4.30-7.94)and 5.01 g/kg.bw(3.08-8.14),respectively.The SBBP belongs to a non-toxic substance.Genetic toxicity testsIn accordance with method of plate incorporation in Bacterial reverse mutation test,the mutagenicity of SBBP was performed with each qualified Salmonella typhimurium strain TA1535,TA97,TA98,TA100 and TA102 at the different concentrations at 5,1.58,0.50,0.158 and 0.050 mg/plate with or without S9 mixture.Meanwhile,spontaneous recovery group(sterile distilled water),DMSO solvent control group and positive control group were also set up.In mammalian cell micronucleus test,fifty healthy ICR mice weighing 25-30 g were randomly divided into 5 groups(10 animals per a group,half male and female).According to individual LD50,these animals were by oral gavage administrated with SBBP twice by 24-hour interval at the doses of 626.25,1252.5,2505 mg/kg.bw for female and 730,1460,2920 mg/kg.bw for male.At the same time,the control group(distilled water)and positive control group(cyclophosphamide,50 mg/kg.bw)were taken in the same way.In mammalian spermatocyte chromosome aberration test,twenty healthy ICR mice weighing 30-35 g were divided into 5 groups,i.e.three treatment groups at 730,1460 and 2920 mg/kg.bw of SBBP,negative control group(distilled water)and positive control group(cyclophospamide,40 mg/kg.bw).The positive control group was only given a single intraperitoneal injection on the first day.Other groups were administered with SBBP by gavage once a day for 5 consecutive days.The 5 mg/kg.bw colchicine was intraperitoneally injected on 13-day.At 4 h after the injection,all groups were sacrificed by cervical dislocation.In conclusion,the results of bacterial reverse mutation test,mammalian cell micronucleus test and mammalian spermatocyte chromosome aberration test are negative,suggesting that the mutagenicity of SBBP is not observed.Repeated dose 28-day oral toxicity studyThirty healthy SD male rats and forty-two female rats were evenly assigned to six groups including the control group,three treatment groups and two additional satellite groups relative to the control group and high dose group.Six experimental groups were treated by gavaged with SBBP at corresponding dose once a day for 28-day but two satellite groups were observed without SBBP for another 14-day.There is no mortality.Under the experimental conditions,general activity,food intake and utilization rate and haematological,clinical biochemical and urinal analysis and main organ weight for rats treated with SBBP up to 2.5 g/kg.bw are no pronounced significance compared to the control group.However,the high dose of SBBP reduces body weight gain of male rats.There are no abnormalities in the gross necropsy and in the histopathology of testes and liver and kidney for female rats except the dose-response dilatation and congestion in hepatic sinus and interrenal hemorrhage for male rats treated with high dose.The no-observed-adverse-effect-level(NOAEL)of SBBP for SD rats is more than 1.25 g/kg.bw in the repeated dose 28-day oral toxicity test.