The Application Of Original Sanitary Inspection Techniques For Determination Of Diethylstilbestrol And Dienestrol In Pharmaceutical And Hair Growth Product

Diethylstilbestrol and dienestrol are synthetic non-steroid phenolic estrogens with similar structures. They are clinically used as supplements for insufficient estrogen and for the treatment of breast cancer, prostatic cancer, amenorrhea and menopausal syndrome. They can also promote the normal development of female genital organs and secondary sexual characters, the propagation of the endometrium, the uterine contraction and the sensitivity to uterus oxytocic hormone. They can stimulate the anterior pituitary to secrete gonodotropin and prolactin in small dosages. However, inhibition effects can be achieved in large dosages. Dienestrol is the in vivo metabolite of diethylstilbestrol. In animal husbandry, diethylstilboestrol and dienestrol are mainly used to increase milk production and promote the synthesis of proteins. They can promote the sedimentation of proteins and reduce the sedimentation of fat. Climacteric disturbance, loss of fertility, sexual precocity in females, feminization in males, breast cancer and oophoroma can be caused after long term use of Stilboestrol and dienestrol or intake of diethylstilboestrol and dienestrol containing milk products or meat products. So it is of high significance to establish convenient and rapid detection methods on Stilboestrol and Dienestrol.Through large amounts of preliminary studies it has been found that luminol can be oxidized by Ag(â…¢) complexes to generate stable chemoluminescence signals in basic media. Some substances can significantly enhance or inhibit the chemoluminescence from luminol-Ag(â…¢) complexes. And there is a good linear relation between strength of chemoluminescence and substance concentration within a certain range. In this study we combined a new luminol-Ag(â…¢) complex chemoluminescence system and a kind of flow injection technology to establish a new method for the detection of diethylstilboestrol and used this method for the assay of diethylstilboestrol tablets.Capillary electrophoresis combines electrophoretic technologies and modern microcolumn separation technologies. With high voltage direct current electric field as the drive, highly efficient and rapid separation can be achieved in capillaries according to different mobility and distribution behaviors of different components. The separation channels for capillary electrophoresis are hollow capillaries. The requirements on pre-treatment of samples are low. The purpose of this study is to establish a capillary electrophoresis method for the detection of Stilboestrol and Dienestrol in hair growth promotion products.PART 1 Detection of diethylstilboestrol content in diethylstilboestrol tablets with a new flow injection - Ag(â…¢) - Luminol chemoluminescence systemObjective: To establish a detection of diethylstilboestrol content in diethylstilboestrol tablets with a novel flow injection - Ag(â…¢) - Luminol chemoluminescence system.Methods: In alkaline medium, a novel flow injection injection - Ag(â…¢) - Luminol chemiluminescent method has been developed for the determination of diethylstilbestrol, based on the inhibition of diethystilbestol on the chemiluminescence reaction between luminol and Ag(III) coordination compounds, and the chemiluminescent intensity show good linearity to the concentration of the analytes within a certain range. Conditions of this chemiluminescent reaction have been systematically investigated and optimized. And this method has been applied for the determination of diethylstilbestrol in diethylstilbestrol tablet.Results: A novel flow injection - Ag(â…¢) - Luminol chemoluminescence system for determination of diethylstilbestrol was developed. The detection limit was 3.0×10^-7 mol/L. The linear equation of the method wasâ–³I=5.84×10⁸C+548, r =0.9989, the recoveries of standard addition was in rang of 88.6%-94.0% . The relative standard deviation was 3.4% for 1.0×10^-5 mol/L of diethylstilbestrol (n=7).Conclusions: The novel flow injection - Ag(â…¢) - Luminol chemoluminescence system can be applied for the determination of diethylstilbestrol in diethylstilbestrol tablet . The method is simple, fast, accurate and reliable. It can be used for fast quality control in process of pharmaceutical production.PART 2 Determination of diethylstilbestrol and dienestrol in hair growth product by capillary electrophoresisObjective: To establish a new method for determination of diethylstilbestrol and dienestrol in hair growth product by capillary electrophoresis with UV detection.Methods: An uncoated fused-silica capillary column (75μm i.d.×55 cm, effective length 50 cm) was used as separation channel. The running buffer contained 20mmol/L borate with additive of 30% (by volume) N,N-Dimethylformamide. A constant voltage of 20 kV was applied and the UV detector was set at 254 nm. Samples were injected by gravity for 10 s.Results: A good baseline separation of diethylstilbestrol and dienestrol was obtained with in 9 min. The calibration curves were linear in the range of 2.0×10^-6-2.0×10^-4mol/L for diethylstilbestrol and 2.0×10^-6-2.0×10^-4mol/L for dienestrol, with linear correlation coefficients of 0.9991 and 0.9994, respectively. The limits of detection of diethylstilbestrol and dienestrol were 6.7×10^-7mol/L and 3.3×10^-7mol/L, respectively. The average recovery of diethylstilbestrol and dienestrol were between 89.6-94.6% and 88.7-104.8%, with the relative standard deviation (RSD) of 3.97% and 4.54%.Conclusion: The method is simple, convenient and sensitive, which is suitable for determination of diethylstilbestrol and dienestrol residues in hair growth product.