Evaluation of Analytical Chemical Methods for Detection of Estrogens in the Environment

Compounds Samples Sample preparation Analytical method LOD/LOQ Ref E1-3S, E1-3G, E2-3S, E2-3G, E2-17S, E2- 3S17G, E2- 3G17S, E2- 3,17DiS, E3-3S, E3-3G, E1, α-E2, ß-E2, E3, EE2 River, lake water, effluent (STP) Detection:   E1-3S: 0.3-2.2 ng/L E2-3S: 0.2-1 ng/L Filtration (1L) pH-adjustment to 3.5-5 with acetic acid SPE: Autoprep EDS-1 Elution: Free: ethyl acetate;          Conjugates: 5mM TEA in MeOH Florisil clean-up (free) Dissolved in 100 µL-1mL LC-MS/MS, ESI (-) StP: Zorbax Extend-C18 column (150 mm x 1 mm I.D., 3.5 µm, Agilent) MP: A: Acetonitrile; B: H2O, C: 100 mM TEA in H2O (pH 12.2)   Temp: 30 ºC, FR: 40µL/min; IV: 10 µL MDL:   Free: 0.1-1.5 ng/L; Conjugates: 0.1-3.1 ng/L (105) E1-3S, E1-3G, E2-3S, E2-3G, E2-17G, E3-3S,   E3-3G,E3-16G   E1, α-E2, ß-E2, E3, EE2 Female urine, septic tank collecting domestic wastewater, influent and effluent (STP) Preservation with formaldehyde (1%, v/v) Filtration (Whatman GF/C glass fiber, 1.5 µm), washing filter with 3 mL MeOH, added to the aqueous extract   SPE (0.5 g Carbograph 4, GCB) washing   After loading of the water samples, the cartridges were washed sequentially with: 50 mL of distilled water, 10 mL MeOH acidified with formic acid 50 mM and 5 mL MeOH Elution of the free: 12 mL methylene chloride:MeOH ( 80:20, v/v) After elution of the free estrogens from the SPE the cartridges were reversed(?), eluted with 20 mL methylene chloride/MeOH (60:40, v/v) containing 10 mM NaAcetate Reconstituted with 200 µL H2O:MeOH (50:50, v/v) after solvent evaporation LC-MS/MS, ESI (-) StP: Alltima C18 column (25x4.6 mm I.D., 3.5 µm) MP: A Acetonitril: B: Water both 10 mM formic acid FR: 1 mL/min; IV: 50 µL LOQ (ng/L): Urine: Free: 16-40; conjugates: 20-600; Wastewater: F:2-4; C: 2- 60;   STP influent: F: 1-2; C: 0.8-30;   STP effluent: F: 0.5-1; C: 0.3-12 (106;107) E1-3S, E1-3G, E2-3S, E2-3G, E2-17G, E3-3S,   E3-3G,E3-16G   Raw sewage, treated sewage, river water Filtration (Whatman GF/C glass fiber, 1.5 µm), washing filter with 3 mL MeOH, added to the aqueous extract   SPE (0.5 g Carbograph 4, GCB) cartridges LC-MS/MS, ESI (-) StP: Alltima C18 column (250 x 4.6 mm I.D., 3.5 µm) LOD (ng/L): STP influent (100 mL) F: 0.4-0.85, C: 0.2-15 STP effluent (250 mL) (108)
E1, α-E2, ß-E2, E3, EE2 washing with 10 mL dichloromethane:MeOH (80:20, v/v) containing 5mM TMACI (tetraethylammonium Chlorid), 5 mL MeOH, 20 mL water acidified (pH 2) with HCl, distilled water 5 mL Loading on the SPE After the loading the SPE were sequentially washed with 50 mL distilled water, 10 mL MeOH acidified with 50 mM formic acid and 10 mL MeOH Elution: F: 10 mL dichloromethane: MeOH (80:20, v/v) into glass vials with conical bottom; C: were back eluted from the SPE with 10 mL dichloromethane: MeOH (80:20, v/v) containing 5mM TMACl into glass vials. Extracts were evaporated at 30 ºC under N2. Re-dissolved in 200 µL Water:MeOH (50:50; v/v) MP: Free: A: Acetonitril: B: Water both 10 mM formic acid; Post column addition of MeOH containing 40 mM NH3, at FR:0.1mL/min Conjugated: Acetonitril: B: Water both 10 mM formic acid; FR: 1 mL/min;   IV: 50 µL F: 0.04-024, C: 0,04-6 River water (2 L) F: 0.005-0.03, C: 0.05-0 E1-3S,E32-3S, E3-3S, E2-17S, E2-3,17DiS d4-E2-3S (IS) Human urine Robotic SPE extraction in the 96.Well Disk Plate LC-MS/MS, ESI (-) StP: Keystone Betasil C18 column (150 x 2 mm, 5 µm) MP: 5 mM ammonium sacetate, pH = 5.4/MeOH (50:50) FR: 200 µL/min; IV: 20 µL LOQ: C: 0.2 ng/mL (109) Human urine HPLC-UV   (110) FR: flow rate; MDL: method detection limit; IV: injection volume; MP: mobile phase; StP: stationary phase.