Building comprehensive glucosinolate profiles for brassica varieties.

Brassica plants play an important role in common agricultural practices, such as livestock feed or biofumigation, due to the bioactivity of the natural degradation products of glucosinolate metabolites. Therefore, the ability to survey comprehensive glucosinolate profiles for individual brassicas is essential for informing proper species selection for the intended application. Current methods for glucosinolate identification and quantification involve complex or unconventional procedures, and proper reference materials are not readily available. Therefore, researchers with limited resources that require glucosinolate profiles are at an extreme disadvantage. In this work, a simple and accurate HPLC-MS method was developed and validated to build preliminary glucosinolate profiles for three agriculturally relevant forage brassica varieties [turnip (B. rapa L.), canola (B. napus L.), and rapeseed (B. napus L.)]. The average glucosinolate content across three herbage collection dates for canola, rapeseed and turnip were 2.9 ± 0.9 mg g-1, 6.4 ± 1.3 mg g-1, and 14 ± 3.4 mg g-1, respectively. GLS concentrations are reported in milligrams of glucosinolate, calculated as sinigrin equivalents, per gram of dry plant material. This semi-quantitative approach for reporting total GLS content in brassicas is accurate within 15%. Several minor individual glucosinolates were identified that have not been previously reported in canola, rapeseed and turnip species, including glucotropaeolin and 4-hydroxyglucobrassicin (canola), glucoraphanin and glucoberteroin (rapeseed), and glucosinalbin and glucobarbarin (turnip). This non-targeted screen of several forage brassica varieties demonstrates the inherent variation in both the individual glucosinolate content and the total glucosinolate profile among brassicas, and highlights the importance of such glucosinolate characterization in agricultural practices. Additionally, the method developed in this study can be used as a tool for researchers with limited resources to build accurate glucosinolate profiles of brassica plants.

Impacts of the COVID-19 pandemic on pharmaceuticals in wastewater treated for beneficial reuse: Two case studies in central Pennsylvania.

During the COVID-19 pandemic, wastewater surveillance was leveraged as a powerful tool for monitoring community-scale health. Further, the well-known persistence of some pharmaceuticals through wastewater treatment plants spurred concerns that increased usage of pharmaceuticals during the pandemic would increase the concentrations in wastewater treatment plant effluent. We collected weekly influent and effluent samples from May 2020 through May 2021 from two wastewater treatment plants in central Pennsylvania, the Penn State Water Reclamation Facility and the University Area Joint Authority, that provide effluent for beneficial reuse, including for irrigation. Samples were analyzed for severe acute respiratory syndrome coronavirus 2 (influent only), two over-the-counter medicines (acetaminophen and naproxen), five antibiotics (ampicillin, doxycycline, ofloxacin, sulfamethoxazole, and trimethoprim), two therapeutic agents (remdesivir and dexamethasone), and hydroxychloroquine. Although there were no correlations between pharmaceutical and virus concentration, remdesivir detection occurred when the number of hospitalized patients with COVID-19 increased, and dexamethasone detection co-occurred with the presence of patients with COVID-19 on ventilators. Additionally, Penn State decision-making regarding instruction modes explained the temporal variation of influent pharmaceutical concentrations, with detection occurring primarily when students were on campus. Risk quotients calculated for pharmaceuticals with known effective and lethal concentrations at which 50% of a population is affected for fish, daphnia, and algae were generally low in the effluent; however, some acute risks from sulfamethoxazole were high when students returned to campus. Remdesivir and dexamethasone persisted through the wastewater treatment plants, thereby introducing novel pharmaceuticals directly to soils and surface water. These results highlight connections between human health and water quality and further demonstrate the broad utility of wastewater surveillance.

Comparison of POCIS and grab sampling techniques for monitoring PPCPs in vernal pools in central Pennsylvania.

Active ingredients in pharmaceuticals and personal care products (PPCPs) can persist through wastewater treatment plants and be released into the environment where they can inadvertently pose risks to non-target organisms. Emerging contaminants (ECs), including PPCPs, are commonly detected in wastewater effluent. With the increasing beneficial re-use of treated wastewater globally, there is a need to understand how spray-irrigation activities affect the occurrence and persistence of ECs in the environment to which they are introduced. Here, we explore the impacts of wastewater spray-irrigation on nearby ephemeral wetlands (e.g., vernal pools) through the use of grab and Polar Organic Chemical Integrative Sampling (POCIS) techniques. This study sought to determine whether integrative sampling techniques are better suited than traditional grab sampling techniques in assessing the presence and concentrations of ECs in vernal pools by evaluating 34 ECs in six vernal pools in central Pennsylvania. Three pools were impacted by wastewater spray-irrigation activities and three were in a nearby forested area. Results of this study found that POCIS detected a wide range of 25 ECs (log Kow between -2.6 and 9.37) more or, in some cases, equally frequently, relative to grab samples. Additionally, grab samples were found to best capture short-lived elevated inputs of ECs (from irrigation events) while POCIS were found to best capture ECs that were present in vernal pools over a longer period of time (weeks to months). For ECs detected more frequently in grab samples, concentrations were higher compared to time weighted average aqueous concentrations estimated from POCIS. This study advances understanding of the potential impact of wastewater beneficial reuse on vernal pools and informs how best to monitor the presence of ECs in vernal pools using integrative and grab sampling techniques.

Small-Grain Cover Crops Have Limited Effect on Neonicotinoid Contamination from Seed Coatings.

Neonicotinoids from insecticidal seed coatings can contaminate soil in treated fields and adjacent areas, posing a potential risk to nontarget organisms and ecological function. To determine if cover crops can mitigate neonicotinoid contamination in treated and adjacent areas, we measured neonicotinoid concentrations for three years in no-till corn-soybean rotations, planted with or without neonicotinoid seed coatings, and with or without small grain cover crops. Although neonicotinoids were detected in cover crops, high early season dissipation provided little opportunity for winter-planted cover crops to absorb significant neonicotinoid residues; small grain cover crops failed to mitigated neonicotinoid contamination in either treated or untreated plots. As the majority of neonicotinoids from seed coatings dissipated shortly after planting, residues did not accumulate in soil, but persisted at concentrations below 5 ppb. Persistent residues could be attributed to historic neonicotinoid use and recent, nearby neonicotinoid use. Tracking neonicotinoid concentrations over time revealed a large amount of local interplot movement of neonicotinoids; in untreated plots, contamination was higher when plots were less isolated from treated plots.

Assessing surface and subsurface transport of neonicotinoid insecticides from no-till crop fields.

Increased use of neonicotinoid-coated crop seeds introduces greater amounts of insecticides into the environment, where they are vulnerable to transport. To understand the transport of neonicotinoids from agricultural fields, we planted maize (Zea mays L.) seeds coated with thiamethoxam in lysimeter plots in central Pennsylvania. Over the next year, we sampled water generated by rainfall and snowmelt and analyzed these samples with mass spectrometry for the neonicotinoids thiamethoxam and clothianidin (metabolite), which originated from the coated seeds. For surface and subsurface transport, thiamethoxam exhibited "first-flush" dynamics, with concentrations highest during the first events following planting and generally decreasing for the remainder of the study. The metabolite clothianidin, however, persisted throughout the study. The mass of thiamethoxam and clothianidin exported during the study period accounted for 1.09% of the mass applied, with more than 90% of the mass transported in subsurface flow and less than 10% in surface runoff. These results suggest that surface runoff, at least for our site, is a relatively small contributor to the overall fate and transport of these insecticides and that the delivery ratio (i.e., mass exported/mass applied) observed for these compounds is similar to those of other trace-level emerging contaminants known to negatively influence aquatic ecosystems.

Method to Evaluate the Age of Groundwater Inputs to Surface Waters by Determining the Chirality Change of Metolachlor Ethanesulfonic Acid (MESA) Captured on a Polar Organic Chemical Integrative Sampler (POCIS).

We previously discovered a method to estimate the groundwater mean residence time using the changes in the enantiomeric ratio of metolachlor ethanesulfonic acid (MESA), (2-[(2-ethyl-6-methylphenyl)(2-methoxy-1-methylethyl)amino]-2-oxoethanesulfonic acid), a metabolite of the herbicide metolachlor. However, many grab samples would be needed for each watershed over an extended period, and this is not practical. Thus, we examined the use of a polar organic chemical integrative sampler (POCIS) deployed for 28 days combined with a modified liquid chromatography-mass spectrometry LC-MS/MS method to provide a time-weighted average of the MESA enantiomeric ratio. POCISs equipped with hydrophilic-lipophilic-balanced (HLB) discs were deployed at five sites across the United States where metolachlor was used before and after 1999 and compared the effectiveness of the POCIS to capture MESA versus grab samples. In addition, an in situ POCIS sampling rate (Rs) for MESA was calculated (0.15 L/day), the precision of MESA extraction from stored POCIS discs was determined, and the effectiveness of HLB to extract MESA was examined. Finally, using molecular modeling, the influence of the asymmetric carbon of metolachlor degradation on the MESA enantiomeric ratio was predicted to be negligible. Results of this work will be used in projects to discern the groundwater mean residence times, to evaluate the delivery of nitrate-N from groundwater to surface waters under various soil, agronomic, and land use conditions, and to examine the effectiveness of conservation practices.

Influence of hydrologic and anthropogenic drivers on emerging organic contaminants in drinking water sources in the Susquehanna River Basin.

Occurrence of emerging organic contaminants (EOCs) in surface water bodies can cause adverse effects on non-target organisms. When surface waters are used as drinking water sources, temporal variability in EOC concentrations can potentially impact drinking water quality and human health. To better understand spatiotemporal variability of EOCs in drinking water sources in Central Pennsylvania, EOCs were evaluated in six drinking water sources during a two-year study period (April 2016-June 2018) in the Susquehanna River Basin (SRB). The study was conducted in two phases: Phase I was a spatially distributed sampling approach within the SRB focusing on seven human pharmaceuticals and Phase II was a temporally intensive sampling regime at a single site focusing on a broader range of EOCs. Concentration-discharge relationships were utilized to classify EOC transport dynamics and understand the extent to which hydrologic and anthropogenic factors, such as surface runoff and wastewater effluent, may contribute to EOC occurrence. Overall, EOCs were present at higher concentrations in colder seasons than warmer seasons. Thiamethoxam, a neonicotinoid insecticide, and caffeine exhibited accretion dynamics during high-flow periods, suggesting higher transport during surface runoff events. Human pharmaceuticals known to persist in wastewater effluent were inversely correlated with discharge, indicating dilution characteristics consistent with diminished wastewater signals during high-flow periods. Acetaminophen exhibited near-chemostatic transport dynamics, indicating nonpoint source inputs during high-flow periods. Risk calculations revealed that although EOCs posed medium-to-high risk to aquatic organisms, human health risk through fish consumption was low.

Occurrence, Concentrations, and Risks of Pharmaceutical Compounds in Private Wells in Central Pennsylvania.

Over-the-counter and prescription medications are routinely present at detectable levels in surface and groundwater bodies. The presence of these emerging contaminants has raised both environmental and public health concerns, particularly when the water is used for drinking either directly or with additional treatment. However, the frequency of occurrence, range of concentrations, and potential human health risks are not well understood, especially for groundwater supplies. Private wells are often not tested for contaminants regulated by drinking water standards and are even less frequently tested for emerging contaminants. By partnering with the Pennsylvania Master Well Owner Network, water samples were collected from 26 households with private wells in the West Branch of the Susquehanna River basin in central Pennsylvania in winter 2017. All samples were analyzed for six pharmaceuticals (acetaminophen, ampicillin, naproxen, ofloxacin, sulfamethoxazole, and trimethoprim) and one over-the-counter stimulant (caffeine). At least one compound was detected at each site. Ofloxacin and naproxen were the most and least frequently detected compounds, respectively. Concentrations from the groundwater wells were higher than those of nearby surface water samples. However, risk calculations revealed that none of the concentrations measured in groundwater samples posed significant human health risk. A simple, physicochemical-based modeling approach was used to predict pharmaceutical transport from septic absorption field to groundwater and further elucidate variations in detection frequencies. Findings indicate that although septic tanks may act as contaminant sources for groundwater wells, the human health impacts from trace-level pharmaceuticals that may be present are likely minimal.

Fate of pharmaceuticals in a spray-irrigation system: From wastewater to groundwater.

Land application of wastewater effluent is beneficial for recharging groundwater aquifers and avoiding direct pollutant discharges to surface waters. However, the fate of non-regulated organic wastewater pollutants, such as pharmaceuticals and personal care products (PPCPs), in such wastewater reuse systems is understudied. Here, a 14-month study (October 2016 through December 2017) was conducted to evaluate the fate and potential risks of seven commonly used PPCPs in a local wastewater treatment plant (WWTP) and from 13 groundwater monitoring wells at a spray-irrigation site where effluent has been spray-irrigated since the early 1980s. Acetaminophen and trimethoprim were the most frequently detected (93%) PPCPs in WWTP influent, while in the effluent, caffeine and trimethoprim were detected most frequently (70%). Wastewater treatment generally reduced concentrations of acetaminophen and caffeine by >88%; however, some compounds had low removal or were present at higher concentrations in the effluent compared with influent (e.g. naproxen, sulfamethoxazole, trimethoprim and ofloxacin). Seasonal trends were observed, with higher PPCP concentrations in the WWTP influent and effluent in the winter. Risk calculations conducted on the wastewater effluent suggest that the risk posed by PPCPs that persisted in the effluent are medium to high to aquatic organisms. Detection frequencies of PPCPs were lower in groundwater samples compared to the effluent, with sulfamethoxazole (40%) and caffeine (32%) as the most frequently detected compounds. Similarly, average concentrations of PPCPs in groundwater were found to be nearly two orders of magnitude lower than concentrations in the effluent. Minimal seasonal influence was observed for groundwater samples. Human health risk assessments indicate that concentrations in groundwater, which is used as a drinking water source, appear to pose minimal risk.

Characterizing the phosphorus forms extracted from soil by the Mehlich III soil test.

Phosphorus (P) can limit crop production in many soils, and soil testing is used to guide fertilizer recommendations. The Mehlich III (M3) soil test is widely used in North America, followed by colorimetric analysis for P, or by inductively coupled plasma-based spectrometry (ICP) for P and cations. However, differences have been observed in M3 P concentrations measured by these methods. Using 31P nuclear magnetic resonance (P-NMR) and mass spectrometry (MS), we characterized P forms in M3 extracts. In addition to the orthophosphate that would be detected during colorimetric analysis, several organic P forms were present in M3 extracts that would be unreactive colorimetrically but measured by ICP (molybdate unreactive P, MUP). Extraction of these P forms by M3 was confirmed by P-NMR and MS in NaOH-ethylenediaminetetraacetic acid extracts of whole soils and residues after M3 extraction. The most abundant P form in M3 extracts was myo-inositol hexaphosphate (myo-IHP, phytate), a compound that may not contribute to plant-available P if tightly sorbed in soil. Concentrations of myo-IHP and other organic P forms varied among soils, and even among treatment plots on the same soil. Extraction of myo-IHP in M3 appeared to be linked to cations, with substantially more myo-IHP extracted from soils fertilized with alum-treated poultry litter than untreated litter. These results suggest that ICP analysis may substantially over-estimate plant-available P in samples with high MUP concentrations, but there is no way at present to determine MUP concentrations without analysis by both colorimetry and ICP. This study also tested procedures that will improve future soil P-NMR studies, such as treatment of acid extracts, and demonstrated that techniques such as P-NMR and MS are complimentary, each yielding additional information that analysis by a single technique may not provide.

Enteric methane production and ruminal fermentation of forage brassica diets fed in continuous culture.

The aim of the current study was to determine nutrient digestibility, VFA production, N metabolism, and CH4 production of canola (Brassica napus L.), rapeseed (B. napus L.), turnip (Brassica rapa L.), and annual ryegrass (Lolium multiflorum Lam.) fed with orchardgrass (Dactylis glomerata L.) in continuous culture. Diets were randomly assigned to fermentors in a 4 × 4 Latin square design using 7 d for adaptation and 3 d for collection. Diets were: 1) 50% orchardgrass + 50% annual ryegrass (ARG); 2) 50% orchardgrass + 50% canola (CAN); 3) 50% orchardgrass + 50% rapeseed (RAP); and 4) 50% orchardgrass + 50% turnip (TUR). Feedings (82 g DM/d) occurred four times daily throughout 4, 10-d periods at 730, 1030, 1400, and 1900 h. Methane samples were collected every 10 min using a photoacoustic gas analyzer (LumaSense Technologies, Inc.; Santa Clara, CA) during the last 3 d of the experiment. Effluent samples were collected on d 8, 9, and 10, composited by fermentor, and analyzed for VFA and pH as well as DM, OM, CP, and fiber fractions for determination of nutrient digestibility. Forage samples were analyzed for CP, NDF, ADF, minerals, and glucosinolate (GLS) concentrations. Data were analyzed using the GLIMMIX procedure of SAS. Apparent DM, OM, and NDF digestibilities and true DM and OM digestibilities were similar (P > 0.28) among diets (45.1, 63.2, 44.1, 67.1, and 87.2%, respectively). Total VFA (87.2 mol/100 mol), pH (6.47), and acetate (A: 44.6 mol/100 mol) were also not different (P > 0.20) among diets. The A:P (P = propionate) ratio was greater (P < 0.01) in ARG and CAN than RAP and TUR. Daily CH4 production was greater (P < 0.01) in ARG than all other diets (68.9 vs. 11.2 mg/d). Methane, whether expressed as g per g of OM, NDF, digestible OM, or digestible NDF fed was greatest (P < 0.01) in ARG but similar (P > 0.18) among brassica diets. A significant negative correlation was observed between total GLS and CH4 production. However, when multiple regression analysis on CH4 production was completed, neither total GLS nor individual GLS were a significant component of the model. Addition of brassicas provided similar nutrient digestibility to ARG while reducing daily CH4 production, potentially making brassicas an alternative for ARG in pasture-based ruminant diets.

Temperature and Nitrogen Effects on Phosphorus Uptake by Agricultural Stream-Bed Sediments.

Climate change will likely increase the growing season, temperatures, and ratio of nitrogen (N) to phosphorus (P) loss from land to water. However, it is unknown how these factors influence P concentrations in streams. We sought to evaluate differences in biotic and abiotic processes affecting stream sediment P dynamics under different temperature and N-enrichment regimes. Three sediments of varying P composition and sorption characteristics were placed into a fluvarium. Synthetic runoff water, with or without added N, was added to the flume's reservoir, and the solution was maintained at 19 or 26°C. Water and sediment samples were taken with time since runoff was introduced. The rate and magnitude of P uptake by sediment was greater at 19°C compared with 26°C, and also when N was added compared with no N added. Analysis of sediment samples indicated that P uptake via abiotic processes was greater at 19 than at 26°C. The addition of N stimulated P uptake by the microbial biomass at 19°C, but microbial uptake was potentially inhibited at 26°C. Because microbial biomass is a temporary store of P, these data suggest that more P may be available with increasing temperatures during the growing season, especially under baseflow, implying that strategies to mitigate P losses from land to water should be strengthened to prevent potential water quality impairment.

Managing Surface Water Inputs to Reduce Phosphorus Loss from Cranberry Farms.

Cranberry ( Ait.) production in Massachusetts represents one-fourth of the US cranberry supply, but water quality concerns, water use, and wetland protection laws challenge the future viability of the state's cranberry industry. Pond water used for harvest and winter flooding accounts for up to two-thirds of phosphorus (P) losses in drainage waters. Consequently, use of P sorbing salts to treat pond water holds promise in the mitigation of P losses from cranberry farms. Laboratory evaluation of aluminum (Al)-, iron (Fe)-, and calcium (Ca)-based salts was conducted to determine the application rate required for reducing P in shallow (0.4 m) and deep (3.2 m) water ponds used for cranberry production. Limited P removal (<22%) with calcium carbonate and calcium sulfate was consistent with their relatively low solubility in water. Calcium hydroxide reduced total P up to 49%, but increases in pond water pH (>8) could be detrimental to cranberry production. Ferric sulfate and aluminum sulfate applications of 15 mg L (ppm) resulted in near-complete removal of total P, which decreased from 49 ± 3 to <10 µg P L (ppb). However, ferric sulfate application lowered pH below the recommend range for cranberry soils. Field testing of aluminum sulfate demonstrated that at a dose of 15 mg L (∼1.4 Al mg L), total P in pond water was reduced by 78 to 94%. Laboratory and field experiments support the recommendation of aluminum sulfate as a cost-effective remedial strategy for reducing elevated P in surface water used for cranberry production.

Reducing Phosphorus Runoff and Leaching from Poultry Litter with Alum: Twenty-Year Small Plot and Paired-Watershed Studies.

Treating poultry litter with alum has been shown to lower ammonia (NH) emissions and phosphorus (P) runoff losses. Two long-term studies were conducted to assess the effects of alum-treated poultry litter on P availability, leaching, and runoff under pasture conditions. From 1995 to 2015, litter was applied annually in a paired watershed study comparing alum-treated and untreated litter and in a small plot study comparing 13 treatments (an unfertilized control, four rates of alum-treated litter, four rates of untreated litter, and four rates of NHNO). In the paired watershed study, total P loads in runoff were 231% higher from pasture receiving untreated litter (1.96 kg P ha) than from that receiving alum-treated litter (0.85 kg P ha). In both studies, alum-treated litter resulted in significantly higher Mehlich III P (M3-P) and lower water-extractable P at the soil surface, reflecting greater retention of applied P and lesser availability of that P to runoff or leaching. In soils fertilized with alum-treated litter, M3-P was much higher when analyzed by inductively coupled argon plasma emission spectrometry than by colorimetry, possibly due to the formation of aluminum phytate. Indeed, alum-treated poultry litter leached less P over the 20-yr study: M3-P at 10 to 50 cm was 266% greater in plots fertilized with untreated litter (331 kg M3-P ha) than with alum-treated litter (124 kg M3-P ha). This research provides compelling evidence that treating poultry litter with alum provides short-term and long-term benefits to P conservation and water quality.

Determination of phytate in high molecular weight, charged organic matrices by two-dimensional size exclusion-ion chromatography.

A two-dimensional chromatography method for analyzing phytate or other ionic targets in matrices containing high molecular weight, charged organic species is described. Prior to quantification by anion exchange chromatography, the sample matrix was prepared by size exclusion chromatography, which removed the majority of the matrix. Quantification of phytate on the AS11-HC was sensitive (0.25µM, 0.17mg/l) and reproducible (4.6% RSD) allowing this method to provide baseline separation of phytate from a manure extract within 14min. The method is simple, requiring only sample filtering, reproducible (between-run variation 5% RSD) and linear from 0.38 to 76µM (0.25-50mg/l). The method is suitable for routine determination of phytate in high organic matrices such as manure extracts.

Portable, fully autonomous, ion chromatography system for on-site analyses.

The basic operating principles of a portable, fully autonomous, ion chromatography system are described. The system affords the user the ability to collect and analyze samples continuously for 27 days, or about 1930 injections before needing any user intervention. Within the 13 kg system, is a fully computer controlled autosampling, chromatography and data acquisition system. An eluent reflux device (ERD), which integrates eluent suppression and generation in a single multi-chambered device, is used to minimize eluent consumption. During operation, about 1 µL of water per minute is lost to waste while operating standard-bore chromatography at 0.5 mL min(-1) due to eluent refluxing. Over the course of 27 days, about 100mL of rinse water is consumed, effectively eliminating waste production. Data showing the reproducibility (below 1% relative standard deviation over 14 days) of the device is also presented. Chromatographic analyses of common anions (Cl(-), NO3(-), SO4(2-), PO4(3-)), is accomplished in under 15 min using a low backpressure guard column with ∼ 25 mM KOH isocratic elution. For detection, a small capacitively-coupled contactless conductivity detector (C4D) is employed, able to report analytes in the sub to low micromolar range. Preconcentration of the injected samples gives a 50-fold decrease in detection limits, primarily utilized for in-situ detection of phosphate (LOQ 10 µg L(-1)). Field analyses are shown for multiple on-site analyses of stream water indifferent weather conditions.

Autonomous, waste-free eluent generation and suppression in a single device: electrodialytic eluent reflux for ion chromatography.

Eluent reflux provides a new approach to suppress and reflux (recover) eluent without the continuous generation of chromatographic waste. The current work utilized a device containing ion exchange membranes at the electrodes, in order to prohibit electrolysis gases from entering the eluent stream. Two resin beds (separated by a membrane stack) were responsible for suppressing incoming eluent and regenerating the suppressed eluent to nearly its original concentration after detection. A greater than expected dilution in the eluent concentration was observed as a result of the minor leakage of potassium ions through the anion membrane stack into the electrode chamber. The incomplete recovery of the eluent was offset by the addition of a three port valve (DRV) to regulate eluent concentration. Over 48 h of continuous operation (192 injections), the device's performance was stable (RSD of 0.21% with the three port valve, compared to RSD 3.73% without). The device was able to operate for up to four weeks using 1L of eluent. Chromatograms showing the reproducibility of the device are presented for anions.

Phytochemical investigation and in vitro cytotoxic evaluation of alkaloids from Abuta rufescens.

A phytochemical investigation of Abuta rufescens was performed to authenticate plant material reported previously and to assess the cytotoxicity of the alkaloids obtained from the plant. Three alkaloids which have not previously been reported from this species, two phenolic (subsessiline, an oxoaporphine, and telitoxine, an azafluoranthene) and one non-phenolic (isoimerubrine, a tropoloisoquinoline), were isolated and identified. These alkaloids, along with others previously isolated from this and another Abuta species (grandirubrine, a tropoloisoquinoline), were evaluated for cytotoxic activity against several human cancer cell lines (HCT-116 colon adenocarcinoma, ACHN renal carcinoma, and A549 lung carcinoma). The tropoloisoquinoline alkaloids (grandirubrine, imerubrine, and isoimerubrine) exhibited the greatest cytotoxicity against the cell lines, especially ACHN and HCT-116 cells. The azafluoranthene alkaloid imeluteine exhibited lesser cytotoxicity, as did one of the oxoaporphine alkaloids.

A comparison of hemolysis and red cell mechanical fragility in blood collected with different cell salvage suction devices.

BACKGROUND: Suction pressure is one variable that can affect the efficiency of red blood cell (RBC) recovery for intraoperative autotransfusion. This study compared a constant-suction-pressure system with a device that is expected to minimize hemolysis by automatically varying the suction pressure. STUDY DESIGN AND METHODS: Twenty-two 50-mL reconstituted whole-blood samples were collected from a flat surface with either a constant-pressure suction device (BRAT 2 autologous blood recovery system, COBE Cardiovascular) set to a pressure of -200 mmHg or a variable-pressure suction device (SmartSuction Harmony surgical suction pump, Haemonetics, Inc.). Time of blood collection was recorded and plasma free hemoglobin (PFHb) was spectroscopically measured in the aspirated blood. Postcollection blood was subjected to RBC mechanical fragility test to characterize potential sublethal blood trauma. Relative hemolysis and the mean fragility index (MFI) were calculated. RESULTS: Hemolysis was significantly reduced with the variable-pressure suction compared to the constant-pressure suction (2.17% vs. 3.20%, respectively; p < 0.001). There was no significant difference between both suction devices in either the MFI of RBCs (0.632 vs. 0.673, respectively; p > 0.05) or the collection time at -200 mmHg (54.2 seconds vs. 52.8 seconds, respectively; p > 0.05). CONCLUSIONS: Although the variable-pressure device produced a significant reduction in hemolysis during one-pass blood collection, the clinical significance of this reduction is not clear. In relative terms, the variable-pressure device would recover an extra 10 mL of RBCs for every liter of salvaged RBCs, which is negligible compared to the blood loss in major surgery.