The Research Of Environmental Estrogens In Fruits That Hunan Residents Often Eat

Objective:To establish High performance liquid chromatography-mass/mass (HPLC-MS/MS)method for the simultaneous determination o f8estrogens, including bisphenol A,4-n-nonyphenol, estrone,17β-estradio, estradiol,17a-ethinylestradiol,4-octylpheno and diethylstilbestrol in fruits. To survey the consumption of residents’ fruit in Hunan province and find the pollution level of several environmental estrogens contamination in some fruits. The study could not only provide the basis for the supervision in the future and control of contaminants,but also could provide references for the formulation of hygienic standard.Methods:1875residents and30households from permanent residents in Class3areas of Hunan Province were selected through multistage randomized cluster sampling. Fruits consumption data was collected by three bookkeeping method. A total of216samples including apples, pears, bananas, oranges, lemons and strawberries were collected through simple random sampling from some fruit wholesale market in Hunan. High performance liquid chromatography-mass/mass (HPLC-MS/MS) with solid-phase extraction, liquid-liquid extraction method was established for determination of bisphenol A,4-n-nonylphenol, estrone,17β-estradio), estradiol,17α-ethinylestradiol,4-octylpheno and diethylstilbestrol in fruit samples. Environmental estrogen exposures of residents from all kinds of fruit were used for preliminary estimating the human exposure risk.Results:1.The residents of hunan province in the intake of fresh fruits each day were105.6g, including28.1g of apple,26.4g of pear,18.1g of banana,10.5g of orange,5.1g of lemon,1.4g of strawberry.2.The important parameters which affect the determination efficiency such as the mobile phase,the condition of mass spectrometry and solid phase extraction column were optimized.The optiomal determination conditions were as follows:the samples were extracted with acetonitrile and ultrasonic at25℃25min, then cleaned-up with an Oasis HLB column,and waters ACQUITY UPLCRBEH C18was selected to separate the analytes. Acetonitrile-methanol (4:1, v/v) and0.1%ammonia aqueous solution were used as the mobile phase by gradient elution in the negative mode. Bisphenol A,4-n-nonylphenol, estrone,17β-estradio, estradiol,17α-ethinylestradiol,4-octylpheno and diethylstilbestrol had good linearity in a range of5～100ng/mL. The detection limits were0.17～0.51ng/g. The average recovery ranged from67.77%to110.49%, and the relative standard deviations (RSDs) were1.08%～12.02%.3.BPA was detected in all fruit samples, and the concentration range was1.26±1.53～2.75±2.13ng/g.The contents of E2in apples,pears and oranges were1.21±0.58ng/g,0.51±0.04ng/g,1.03±0.24ng/g respectively, P <0.05significantly statistical different results. The concentration of4-n-NP and EE2in all the fruit samples except strawberries was between2.67±1.47～12.91±9.66ng/g and0.93±0.59～1.52±1.03ng/g respectively. DES, OP, E1and E3were not detected in all samples. The total daily amount of BPA,EE2,E2and4-n-NP intake from the fruits were3.051、1.625、0.925、11.146ng/(kg·bw) respectively. The risk indexes for the six environmental estrogens were lower than1.Conclusions:High performance liquid chromatography-mass/mass (HPLC-MS/MS) method has been established for the simultaneous determination o f8estrogens, including bisphenol A,4-n-nonylphenol, estrone,17β-estradio, estradiol,17α-ethinylestradiol,4-octylpheno and diethylstilbestrol in fruits. The exposure content of environmental estrogens in some fruits was safe. Therefore, the amount of environmental estrogens intakes through the fruits diet were found to be at a relatively safe level. They would not affect adversely to the health of the local residents.