Study On Determination Of Nandrolone Phenylpropionate Added In Two Veterinary Traditional Chinese Herb Medicine By HPLC

Norone phenylpropionate(NP)is a protein assimilating hormone.It is a synthetic steroid compound derived from androgen and increase protein synthesis.In order to promote the growth of animals,NP was added into chinese herbal medicines.The residues of NP in animal food have adverse effects on human health.The long-term use of exogenous androgens produces immunosuppressive effects on the body and causes the disorder of hormone metabolism in the body.In order to detect the NP in chinese herbal medicines,a HPLC method was established to detect the addition of NP in two chinese herbal medicines.The samples were extracted with methanol.The chromatographic conditions were performed on Eclipse Plus C18 column(4.6 mm × 250 mm × 5 ?m)with a mobile phase of acetonitrile-water,gradient elute,a detector wavelength of 241 nm,a flow rate of 1.0 mL/min and a column temperature of 35?.The injection volume was 10 ?L.The results showed a good linear relationship in the linear range of 1.0 ?g/mL?50.0 ?g/mL(R2>0.999).The recovery of the NP added in two chinese herbal medicines were 95.5%and 96.18%,respectively.The intra-assay coefficient of variation were 0.44%?3.67%,0.50%?3.38%respectively,and the coefficient of variation between batches were 0.10%?1.38%,0.02%?0.65%respectively.The coefficient of variation is all less than 10%.The limit of detection was 1.0 mg/kg,and limit of quantitation was 2.0 mg/kg of the NP added in two chinese herbal medicines.The NP concentration of high,medium,low and LOQ in the samples maintained good stability under room temperature condition.Man-machine verification test showed that a good linear relationship in the linear range of 1.0 ?g/mL?50.0 ?g/mL(R2>0.998).The deviation of recovery and coefficient of variation of NP in the two Chinese herbs were within 5%.The deviation of LOD and LOQ of variation of NP in man-machine verification test were within 5%.The established detection method was used to detect NP in six samples of two batches collected from 2017 to 2018 years.The results showed that NP was illegally added to one sample with 1.0 mg/kg.The established methods had specificity,precision,recovery and stability that meet the criterion of methodology.It is a practical significance to improve the ability to supervise the illegal addition of phenylpropionate in chinese herbal medicine and to ensure the safety of animal food.