Hua-Jie Lai

Trace analysis of 28 steroids in surface water, wastewater and sludge samples by rapid resolution liquid chromatography–electrospray ionization tandem mass spectrometry

A sensitive rapid resolution liquid chromatography-tandem mass spectrometry (RRLC-MS/MS) method, combined with solid-phase extraction, ultrasonic extraction and silica gel cartridge cleanup, was developed for 28 steroids including 4 estrogens (estrone (E1), 17β-estradiol (E2), 17α-ethynyl estradiol (EE2), diethylstilbestrol (DES)), 14 androgens (androsta-1,4-diene-3,17-dione (ADD), 17α-trenbolone, 17β-trenbolone, 4-androstene-3,17-dione, 19-nortestoserone, 17β-boldenone, 17α-boldenone, testosterone (T), epi-androsterone (EADR), methyltestosterone (MT), 4-hydroxy-androst-4-ene-17-dione (4-OHA), 5α-dihydrotestosterone (5α-DHT), androsterone (ADR), stanozolol (S)), 5 progestagens (progesterone (P), ethynyl testosterone (ET), 19-norethindrone, norgestrel, medroxyprogesterone (MP)), and 5 glucocorticoids (cortisol, cortisone, prednisone, prednisolone, dexamethasone) in surface water, wastewater and sludge samples. The recoveries of surface water, influents, effluents and sludge samples were 90.6-119.0% (except 5α-DHT was 143%), 44.0-200%, 60.7-123% and 62.6-138%, respectively. The method detection limits for the 28 analytes in surface water, influents, effluents and freeze-dried sludge samples were 0.01-0.24 ng/L, 0.02-1.44 ng/L, 0.01-0.49 ng/L and 0.08-2.06 ng/g, respectively. This method was applied in the determination of the residual steroidal hormones in two surface water of Danshui River, 12 wastewater and 8 sludge samples from two wastewater treatment plants (Meihu and Huiyang WWTPs) in Guangdong (China). Ten analytes were detected in surface water samples with concentrations ranging between 0.4 ng/L (17β-boldenone) and 55.3 ng/L (5α-DHT); twenty analytes in the wastewater samples with concentrations ranging between 0.3 ng/L (P) and 621 ng/L (5α-DHT); and 12 analytes in the sludge samples with concentrations ranging between 1.6 ng/g (E1) and 372 ng/g (EADR).

Occurrence and fate of eleven classes of antibiotics in two typical wastewater treatment plants in South China

Wastewater treatment plants (WWTPs) are regarded as one of the most important sources of antibiotics in the environment. Two sampling campaigns over a period of one year in two wastewater treatment plants (plant A: activated sludge with chlorination, and plant B: oxidation ditch with UV) of Guangdong Province, China were carried out to assess the occurrence and fate of 11 classes of 50 antibiotics. The wastewater samples were extracted by Oasis HLB cartridges (6mL, 500 mg), while the solid samples (sludge and suspended solid matter) were extracted by ultrasonic-assisted extraction with solvents (acetonitrile and citric acid buffer), followed by an enrichment and clean-up step with solid-phase extraction using SAX-HLB cartridges in tandem. The results showed the presence of 20 and 17 target compounds in the influents and effluents, respectively, at the concentrations ranging from low ng/L to a few μg/L. Sulfamethoxazole, norfloxacin, ofloxacin, anhydro erythromycin and trimethoprim were most frequently detected in the WWTPs wastewaters. Twenty-one antibiotics were found in the sewage sludge from the two WWTPs at the concentrations up to 5800ng/g, with tetracycline, oxytetracycline, norfloxacin and ofloxacin being the predominant antibiotics. The total mass loads of antibiotics per capita in the two plants ranged from 494 to 901 μg/d/inhabitant (672±182 μg/d/inhabitant) in the influents, from 130 to 238 μg/d/inhabitant (175±45 μg/d/inhabitant) in the effluents and from 107 to 307 μg/d/inhabitant (191±87.9 μg/d/inhabitant) in the dewatered sludge, respectively. The aqueous removals for sulfonamides, macrolides, trimethoprim, lincomycin and chloramphenicol in the WWTPs were mainly attributed to the degradation processes, while those for tetracyclines and fluoroquinolones were mainly due to the adsorption onto sludge.

Excretion masses and environmental occurrence of antibiotics in typical swine and dairy cattle farms in China

This paper evaluated the excretion masses and environmental occurrence of 11 classes of 50 antibiotics in six typical swine and dairy cattle farms in southern China. Animal feeds, wastewater and solid manure samples as well as environmental samples (soil, stream and well water) were collected in December 2010 from these farms. Twenty eight antibiotics, including tetracyclines, bacitracin, lincomycin, sulfonamides, fluoroquinolones, ceftiofur, trimethoprim, macrolides, and florfenicol, were detected in the feeds, animal wastes and receiving environments. The normalized daily excretion masses per swine and cattle were estimated to be 18.2mg/day/swine and 4.24 mg/day/cattle. Chlortetracycline (11.6 mg/day/swine), bacitracin (3.81 mg/day/swine), lincomycin (1.19 mg/day/swine) and tetracycline (1.04 mg/day/swine) were the main contributors to the normalized daily excretion masses of antibiotics per swine, while chlortetracycline (3.66 mg/day/cattle) contributed 86% of the normalized daily excretion masses of antibiotics per cattle. Based on the survey of feeds and animal wastes from the swine farms and interview with the farmers, antibiotics excreted by swine were mainly originated from the feeds, while antibiotics excreted by dairy cattle were mainly from the injection route. If we assume that the swine and cattle in China excrete the same masses of antibiotics as the selected livestock farms, the total excretion mass by swine and cattle per annum in China could reach 3,080,000 kg/year and 164,000 kg/year. Various antibiotics such as sulfonamides, tetracyclines, fluroquinolones, macrolides, trimethoprim, lincomycin and florfenicol were detected in well water, stream and field soil, suggesting that livestock farms could be an important pollution source of various antibiotics to the receiving environments.

4-Nonylphenol, bisphenol-A and triclosan levels in human urine of children and students in China, and the effects of drinking these bottled materials on the levels.

4-Nonylphenol (4-NP), bisphenol-A (BPA) and triclosan (TCS) are three industrial chemicals used widely in daily products. This study investigated 4-NP, BPA and TCS levels in urine samples of 287 children and students aged from 3 to 24 years old in Guangzhou, China. Total (free and conjugated) amounts of 4-NP, BPA and TCS in the urine samples were detected using gas chromatography-mass spectrometry with negative chemical ionization. The detection rates of 4-NP, BPA and TCS were 100%, 100% and 93% respectively, given the detection limits of 3.8, 0.5 and 0.9 ng/L respectively. Data for 4-NP, BPA and TCS were presented in both creatinine-adjusted (microgram per gram creatinine) and unadjusted (microgram per liter) urinary concentrations. The geometric mean (GM) concentrations of urinary 4-NP, BPA and TCS were 15.92 μg/g creatinine (17.40 μg/L), 2.75 μg/g creatinine (3.00 μg/L) and 3.55 μg/g creatinine (3.77 μg/L) respectively. Multiple regression models considering age, gender, preferred drinking bottle and log-transformed creatinine were used to calculate the adjusted least square geometric mean (LSGM). Among these subjects, the females had higher LSGM concentrations of 4-NP, BPA and TCS than the males; and the only statistically significant difference was found for the LSGM concentrations of triclosan (p=0.031). Participants who reported to use ceramic cups more frequently had significantly lower LSGM concentrations of BPA than those who used plastic cups (p=0.037). Meanwhile, a three-week test of using polycarbonate bottles and ceramic cups to drink bottled water and boiled tap-water was carried out among 12 graduate students of 25 years old. The GM concentrations of urinary BPA at the end of the first week after using ceramic cups to drink bottled water were 7.16 μg/g creatinine, then decreased significantly to 3.49 μg/g creatinine after the second week of using ceramic cups to drink boiled tap-water (p<0.05), and finally increased to 4.15 μg/g creatinine after the third week of using polycarbonate bottles in drinking boiled tap-water. The results indicate that in daily life the use of polycarbonate bottles or drinking of bottled water is likely to increase the ingestion of BPA, resulting in an increase in urinary BPA levels.

Monitoring of selected estrogenic compounds and estrogenic activity in surface water and sediment of the Yellow River in China using combined chemical and biological tools

We investigated occurrence of selected compounds (4-t-octylphenol: 4-t-OP; 4-nonylphenols: 4-NP; bisphenol-A: BPA; estrone: E1; 17β-estradiol: E2; triclosan: TCS) and estrogenicity in surface water and sediment of the Yellow River in China by using combined chemical analysis and in vitro yeast screen bioassay. Estrogenic compounds 4-t-OP, 4-NP, BPA, E1, E2 and TCS were measured in the water samples, with their average concentrations of 4.7, 577.9, 46.7, 1.3, ND and 6.8 ng/L, respectively. In sediment, the average concentrations of 4-t-OP, 4-NP, BPA and TCS were 35.7, 0.5, 1.7 and 0.7 ng/g while E1 and E2 were not detected in the sediments of all selected sites. In general, the estrogenic compounds in surface water and sediment of the Yellow River were at relatively low levels, thus having medium to minimal estrogenic risks in most sites except for the site of east Lanzhou with high estrogenic risks.

Screening of multiple hormonal activities in surface water and sediment from the Pearl River system, South China, using effect‐directed in vitro bioassays

This paper reports screening of multiple hormonal activities (estrogenic and androgenic activities, antiestrogenic and antiandrogenic activities) for surface water and sediment from the Pearl River system (Liuxi, Zhujiang, and Shijing rivers) in South China, using in vitro recombinant yeast bioassays. The detection frequencies for estrogenic and antiandrogenic activities were both 100% in surface water and 81 and 93% in sediment, respectively. The levels of estrogenic activity were 0.23 to 324 ng 17β-estradiol equivalent concentration (EEQ)/L in surface water and 0 to 101 ng EEQ/g in sediment. Antiandrogenic activities were in the range of 20.4 to 935 × 10(3) ng flutamide equivalent concentration (FEQ)/L in surface water and 0 to 154 × 10(3) ng FEQ/g in sediment. Moreover, estrogenic activity and antiandrogenic activity in sediment showed good correlation (R(2) = 0.7187), suggesting that the agonists of estrogen receptor and the antagonists of androgen receptor co-occurred in sediment. The detection frequencies for androgenic and antiestrogenic activities were 41 and 29% in surface water and 61 and 4% in sediment, respectively. The levels of androgenic activities were 0 to 45.4 ng dihydrotestosterone equivalent concentration (DEQ)/L in surface water, and the potency was very weak in the only detected sediment site. The levels of antiestrogenic activity were 0 to 1,296 × 10(3) ng tamoxifen equivalent concentration (TEQ)/L in surface water and 0 to 89.5 × 10(3) ng TEQ/g in sediment. The Shijing River displayed higher levels of hormonal activities than the Zhujiang and Liuxi rivers, indicating that the Shijing River had been suffering from heavy contamination with endocrine-disrupting chemicals. The equivalent concentrations of hormonal activities in some sites were greater than the lowest-observed-effect concentrations reported in the literature, suggesting potential adverse effects on aquatic organisms.

Fate and occurrence of steroids in swine and dairy cattle farms with different farming scales and wastes disposal systems

Fate and occurrence of fourteen androgens, four estrogens, five glucocorticoids and five progestagens were investigated in three swine farms and three dairy cattle farms with different farming scales and wastes disposal systems in China. Twenty-one, 22, and 12 of total 28 steroids were detected in feces samples with concentrations ranging from below method limit of quantitation (<LOQ for estrone) to 8100 ± 444 ng/g (progesterone), in wastewater samples with concentrations ranging from <LOQ (estrone) to 20,700 ± 1490 ng/L (androsterone), in suspended particles with concentrations ranging from <LOQ (17β-trenbolone) to 778 ± 82.1 ng/g (5α-dihydrotestosterone) in the six farms, respectively. The steroids via swine farms and human sources were mainly originated from wastewater into the receiving environments while those steroids via cattle farms were mainly from cattle feces. The total contributions of steroids to the environment in China are estimated to be 139, 65.8 and 60.7 t/year from swine, dairy cattle and human sources, respectively.

Biotransformation of progesterone and norgestrel by two freshwater microalgae (Scenedesmus obliquus and Chlorella pyrenoidosa): transformation kinetics and products identification.

Natural and synthetic steroid hormones such as progesterone and norgestrel in the aquatic environment may cause adverse effects on aquatic organisms. This study investigated the biotransformation of progesterone and norgestrel in aqueous solutions by two freshwater microalgae Scenedesmus obliquus and Chlorella pyrenoidosa and elucidated their transformation mechanisms. More than 95% of progesterone was transformed by the two microalgae within 5d. For norgestrel, almost complete transformation by S. obliquus was observed after 5 d, but nearly 40% was remained when incubated with C. pyrenoidosa. The results also showed that these two compounds were not accumulated in the algal cells. Biotransformation was found to be the main mechanism for their loss in the aqueous solutions, and it followed the first-order kinetic model. For progesterone, three main transformation products, i.e. 3β-hydroxy-5α-pregnan-20-one, 3,20-allopregnanedione and 1,4-pregnadiene-3,20-dione, and six minor androgens were identified. For norgestrel, only two transformation products, 4,5-dihydronorgestrel and 6,7-dehydronorgestrel, were identified for the first time. Hydroxylation, reduction and oxidation are proposed to be the main transformation pathways. Among the two microalgae species, S. obliquus was found more efficient in the transformation of the two target compounds than C. pyrenoidosa. The results clearly demonstrated the capability of the two microalgae to transform the two progestogens. The biotransformation and products could have significant environmental implications in the fate and effects of the two steroids.

Field dissipation and risk assessment of typical personal care products TCC, TCS, AHTN and HHCB in biosolid-amended soils

The antimicrobial agents triclocarban (TCC) and triclosan (TCS) and synthetic musks AHTN (Tonalide) and HHCB (Galaxolide) are widely used in many personal care products. These compounds may release into the soil environment through biosolid application to agricultural land and potentially affect soil organisms. This paper aimed to investigate accumulation, dissipation and potential risks of TCC, TCS, AHTN and HHCB in biosolid-amended soils of the three field trial sites (Zhejiang, Hunan and Shandong) with three treatments (CK: control without biosolid application, T1: single biosolid application, T2: repeated biosolid application every year). The one-year monitoring results showed that biosolids application could lead to accumulation of these four chemicals in the biosolid-amended soils, with the residual concentrations in the following order: TCC>TCS>AHTN>HHCB. Dissipation of TCC, TCS, AHTN and HHCB in the biosolid-amended soils followed the first-order kinetics model. Half-lives for TCC, TCS, AHTN and HHCB under the field conditions of Shandong site were 191, 258, 336 and 900 days for T1, and 51, 106, 159 and 83 days for T2, respectively. Repeated applications of biosolid led to accumulation of these personal care products and result in higher ecological risks. Based on the residual levels in the trial sites and limited toxicity data, high risks to soil organisms are expected for TCC and TCS, while low-medium risks for AHTN and HHCB.

Biotransformation of the flame retardant tetrabromobisphenol-A (TBBPA) by freshwater microalgae

Tetrabromobisphenol-A (TBBPA) is the most widely used brominated flame retardant. However, little is known about its biotransformation by algae in aquatic environments. The authors investigated transformations of TBBPA by 6 freshwater green microalgae and identified its transformation products. Transformation experiments were conducted under axenic conditions in a laboratory for 10 d. The results showed that TBBPA could be transformed by the selected microalgae, with nearly complete removal by Scenedesmus quadricauda and Coelastrum sphaericum following 10-d incubation. Five transformation products were positively identified by mass spectrometry: TBBPA sulfate, TBBPA glucoside, sulfated TBBPA glucoside, TBBPA monomethyl ether, and tribromobisphenol-A. The mechanisms involved in the biotransformation of TBBPA include sulfation, glucosylation, O-methylation, and debromination, which could be an important step for its further degradation. This suggests that microalgae can play an important role in the fate of TBBPA in aquatic environments. The present study is the first report on algal transformation of TBBPA, and the proposed transformation products could have significant environmental implications.