Determination Study Of Sexual Hormones In Cosmetic By HPLC

Dissertation contents of the thesis were:①Developed2kinds of determination methods of5forbidden sexual hormones in cosmetics by high performance liquid chromatography, including norgestrel, megestrol acetate, chlormadinone acetate, medroxyprogesterone acetate, propionate testosterone.2kinds of pretreatment methods were developed for lotion, emulsion, face cream and essential oils.②Improved the chromatographic detection conditions for the determination of7sexual homones reported in2007"Hygienic Standard for Cosmetics", in-cluding progesterone, testosterone, methyl testosterone, estrone, estradiol, estriol, diethylstil-bestrol.The liquid chromatography instrument conditions of separating5sexual hormones were discussed. The instrument separating parameters including the proportion of the mobile phase, column, column temperature, flow rate, injection volume, detection wavelength selection. The finalized instrument parameters were:Column:Kromasil100-5C18column (250mm×4.6mm,5μm); mobile phase:acetonitrile+water=65+35(v+v); flow rate:1.OmL/min; in-jection volume:5μL; UV detection wavelength:242,290nm; column temperature:25°C The linear plots were obtained between100and10000μg/L with good correlation coeffi-cients larger than0.9990. The limits of detection (LODs) for instrument were18.92-46.78μg/L, the qualification of detection (QOD) were33.1-131.0μg/L;the RSD of intraday and day precision were below4%. The results indicated that the developed method was accurate and suitable.After comparing the effection of different pretreatment methods, such sa solvent extrac-tion, adding precipitating agent, solid-phase column purification, the pretreatment method of5sexual hormones in cosmetics was established.The cosmetic samples were extracted with methanol centrifugal filtration, the solid inorganic salts demulsification injection, after swirled, crushed by ultrasound and centrifugated, then injected after been filtrated. The cos-metics samples were extracted by methanol and demulsified by solid inorganic salt. The lim-its of detection (LODs) for this method in cosmetic were in the range of0.552-1.008mg/Kg, the qualification of detection (QOD) were between1.104-5.040mg/Kg. The limits of with the recoveries were in the range of97-123%. The relative standard deviations (RSD) were below3%. The results indicated that the developed method is simple, rapid, accurate and suitable for the determination of5sexual hormones in cosmetic samples.Based on previous studies, the improvement of pretreatment method to essential oil was discussed. Blank essential oils were spiked by hexane solution,90%methanol in water (v+v) solution extraction, degreased by n-hexane.The essential oil samples were extracted by90%methanol-water (V/V). The detective wavelength was242and290nm. The limits of de-tection (LODs) for this method were in the range of0.550-5.040mg/Kg, the qualification of detection (QOD) were between0.662-5.260mg/Kg. The recoveries were in the range of80-122%except Testosterone propionate. The relative standard deviations (RSD) were below3%. The results indicated that the developed method is simple, rapid, accurate and suitable for the determination of5sexual hormones in essential oil.The chromatography parameters of stimultaneous determinating progesterone, the sex hormones testosterone, methyl testosterone, estrone, estradiol, estriol, diethylstilbestrol which were involved in the "Hygienic Standard for Cosmetics" were improved at the last.7sexual hormones could be separated simultaneously to baseline and reproducted well under2kinds of mobilephase ratio. The effection of changing mobile phase ratio to the chromatographic behavior was analysted, and a curve fitting method of sepration data was developed for sub-sequent determination.