Screening Of Arylsulfatase/?-glucuronidase Inhibitors And Their Application In The Storage Of Estrogen Conjugates Wastewater Samples

It is known that estrogen conjugates in municipal wastewater exist mainly as sulfate and glucuronide conjugates,which can be degraded to their correspongding natural estrogens by arylsulfatase and?-glucurondase.Therefore,it is necessary to keep estrogen conjuagtes intact during sample collection and storage.This study aims to find an effective inhibition condition for arylsulfatase and?-glucurondase.To elucidate these conditions,inhibiton experiments were performed by adding several inhibitors or by introducing extreme p H conditions.This work confirms that arylsulfatase and?-glucurondase can tolerate some extremes,including 1m M Hg Cl₂?1m M EDTA and 9.5%methanol.Low p H?<3?can effectively inhibit arylsulfatase and?-glucurondase,while sulfate conjugates can be degraded by acid.High p H?11?can effectively inhibit arylsulfatase and?-glucurondase and no deconjugation of sulfate and glucuronide conjugates are occurred.Thus,p H of 11 is an apporitate storage condition for the effective inhibiton of arylsulfatase and?-glucurondase.This storage condition was confirmed with diluted activated sludge samples,in which sulfate and glucuronide conjugates haven't been deconjugated within 48 h.In this study,the activity of arylsulfatase and?-glucurondase at different concentration of Hg Cl₂ was texted.Through the dose-carve,it can be found that 2 g/L of Hg Cl₂ has an effective inhibition of two enzymes.Inhibition experiments were performed with diluted activated sludge samples under room temperature?2 g/L of Hg Cl₂?low p H?1??high p H?11?.The result show that sulfate and glucuronide conjugates haven't been deconjugated within 48 h only at a concentration of 2 g/L of Hg Cl₂.In summary,both p H of 11 and 2 g/L of Hg Cl₂ are apporitate storage condition to prevent deconjugation of sulfate and glucuronide conjugates in wastewater during sample collection and storage.However,when there are many microorganisms in the wastewater,it is more appropriate to store the conjugate samples by adding 2 g/L of Hg Cl₂,because the role of microorganisms will make it difficult to maintain the p H of the wastewater at 11.Arylsulfatase and?-glucurondase are two kinds of hydrolytic enzymes wildly present in human body,which are closely related to human health.Studies have found that the activities of arylsulfatase and?-glucuronidase in patients with breast cancer,ovarian cancer,liver cancer and other diseases are significantly higher than those in normal people.Therefore,inhibiting the activity of these two enzymes in patients unusually is a means of treating such cancers.In this paper,the effects of different concentrations of sodium sulfite,hydrogen peroxide,sodium lauryl sulfate,and sodium hypochlorite on the activities of arylsulfatase and?-glucuronidase were studied.Sodium sulfite,hydrogen peroxide,sodium lauryl sulfate,and sodium hypochlorite have a good inhibitory effect on arylsulfatase and the IC₅₀ were 31.51,145.2,181.17,82.66?mol/L,respectively.Sodium sulfite and hydrogen peroxide have no obvious inhibitory effect on?-glucuronidase.Sodium dodecyl sulfate has inhibitory effect on?-glucuronidase but the inhibitory effect is limited,the maximum degree of inhibition is about38%.The IC₅₀ of sodium hypochlorite on?-glucuronidase is 784.4?mol/L.