Removal of Natural Steroid Hormones from Wastewater
Joshua L. Cartinella, Tzahi Y. Cath, Michael T. Flynn, Glenn C. Miller, Kenneth W. Hunter Jr., Amy E. Childress Materials and Methods Solution Chemistry – Wastewater Ersatz
The humidity condensate and urine simulant solutions used in the current investigation
correspond to recipe 1A for Transit Mission Wastewater Ersatz (19) and the hygiene wastewater
simulant solution corresponds to recipe 2A for Early Planetary Base Wastewater Ersatz (19).
Table S1. Wastewater Ersatzs (all purities > 99% unless otherwise noted) Concentrate Concentrate Concentrate 1 (10x): Urine 1 – Organics
Ammonium oxalate monohydrate (NH4)2C2O4 142.10 0.665
Concentrate 2 (10x): Urine 2 – Inorganics Concentrate Concentrate Concentrate 3 (10x): Humidity Condensate Concentrate 4 (20x): Hygiene Wastewater Chemical Pretreatment
1 Geropone TC42 (formerly Igepon TC-42) is manufactured by Rhodia North America Chemicals and is approximately 60% water.
Solid Phase Extraction Procedure
Sep-Pak C18 columns (WAT036905, Waters Corp., Milford, MA) were sequentially
conditioned with 4 mL of HPLC-grade methanol and 6 mL of DDW. Depending on the type of
sample, either 3, 6, 12, or 120 mL of sample were passed through the columns at a rate of 10
mL/min. 15 mL of DDW were then passed through the columns to remove any loosely-bound
components. The columns were dried for 10 min under vacuum before being eluted with 5 mL
of HPLC-grade methanol. The methanol was evaporated under a gentle stream of nitrogen to a
volume of less than 0.2 mL. Buffer provided by the immunoassay kit manufacturers was then
added to the samples to produce a total sample volume of 12 mL, which could be analyzed using
the immunoassay kits. Depending on the initial volume of sample added to the columns, the
final samples (following preparation) were at a dilution/concentration of either 1:4, 1:2, 1:1, or
10:1. Samples had to be diluted/concentrated to ensure that they would fall in the linear range of
the standard curves produced by the kits. An extraction efficiency test was performed to ensure
that a high percentage of hormones were recovered from the initial samples using the extraction
Extraction Efficiencies of Estrone and Estradiol
The extraction procedure was found to remove approximately 85% of the EIA kit
interference experienced when samples were not extracted. The remaining 15% of the
interference was not taken into account for rejection calculations because interference was found
to cause the estrone and estradiol concentrations in the permeate samples to be overstated.
Therefore, interference experienced during the EIA kit analyses, if anything, would have led to
reporting of conservative rejection results. An extraction efficiency test was performed on
samples containing estradiol. Based on results summarized in Table S1, an average extraction
efficiency of 80% was used for rejection calculations. The recoveries summarized in Table S1
were taken from three different extraction tests. The procedure was refined for each extraction
test, and the extraction procedure used in the analysis of the samples reported in this paper
correspond to the procedure with 96.8% recovery. Therefore, using an extraction efficiency of
80% substantiates that conservative rejection values were calculated.
Table S2. Recovery of estradiol from water samples using solid-phase extraction. Concentration Concentration Before Extraction After Extraction % Recovery EIA Kit Analysis for Estrone and Estradiol
Detection limits of 10 ng/L and 8 ng/L for estrone and estradiol, respectively, were given
by the manufacturers and verified independently. Permeate samples were concentrated 10:1 as
described above so that concentrations in the range of 1 ng/L could be detected. The estrone EIA
kit manufacturer reported a 2.2% cross-reactivity with estradiol, and the estradiol EIA kit
manufacturer reported a 4% cross-reactivity with estrone. Cross-reactivity was verified
independently to be negligible for both kits. Both manufacturers reported an intra-assay
coefficient of variation (CV) of 10% which was verified independently.
Results and Discussion Chemical Pretreatment of Waste Streams
Versostko et al. (19) provides a chemical pretreatment procedure for feed solutions
containing urine. The pretreatment is achieved by addition of strong oxidants (potassium
monopersulfate (oxone), potassium benzoate, and sulfuric acid) to the feed solution. The
purpose of the pretreatment is to oxidize organic compounds in the urine and to maintain low pH
levels in the wastewater to prevent the release of ammonia. This is particularly important for the
DCMD process because otherwise, ammonia will evaporate with the water and not be rejected.
In the current investigation, Verostko’s pretreatment was not used because it would likely
cause oxidation of the estrone and estradiol. Therefore, for the feed solutions used in the DCMD
experiments, only sulfuric acid was added to adjust the pH to 3. For the feed solutions used in
the FO experiments, only NaOH was added to adjust the pH to 6.5 to prevent oxidation of the
Although Verostko’s pretreatment was not performed on any of the feed solutions used in
the DCMD and FO experiments, it was important to understand the extent of estrone and
estradiol degradation during the chemical pretreatment. The estrone and estradiol concentrations
were measured before and after chemical pretreatment (i.e. potassium monopersulfate (oxone)
and potassium benzoate addition) of a wastewater feed solution containing humidity condensate
and urine components. After chemical pretreatment, the estradiol concentration decreased by
approximately 48% and the estrone concentration increased by approximately 12%. The
increase in estrone concentration can be attributed to the fact that estrone is a degradation
byproduct of estradiol (24). Therefore, it is estimated that the chemical pretreatment was able to
degrade approximately 36% of hormones overall. The use of this pretreatment for direct potable
use applications in advanced life support system would help lower concentrations of hormones
having to be removed by the main treatment process (e.g., DCMD or FO).
Possible species for the production of biodiesel on marginal land B A McKenzie1, B M Smallfield2, V Fasi1 and R J Martin1 1Bioprotection Research Centre, P O Box 84, Lincoln University, Lincoln 7647, New Zealand 2The New Zealand Institute for Plant & Food Research Limited, Private Bag 4704, Christchurch Abstract A series of field experiments was conducted in 2009-10 and 2010-11 in t
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