Medical waste management plan.

This plan describes the process for managing research generated medical waste at Sandia National Laboratories/California. It applies to operations at the Chemical and Radiation Detection Laboratory (CRDL), Building 968, and other biosafety level 1 or 2 activities at the site. It addresses the accumulation, storage, treatment and disposal of medical waste and sharps waste. It also describes the procedures to comply with regulatory requirements and SNL policies applicable to medical waste

Preservation of solid mercuric dithizonate samples with polyvinyl chloride for determination of mercury(II) in environmental waters by photochromism-induced photoacoustic spectrometry

A novel sample preparation technique has been developed to preserve solid mercuric dithizonate [Hg(HDz)2] in a matrix of polyvinyl chloride (PVC) for analysis by photochromism-induced photoacoustic spectrometry (PCPAS). This technique, which begins with the extraction of Hg2+ from water with dithizone, allows for the determination of Hg2+ in environmental samples. Inclusion of Hg(HDz)2 within the polymer matrix enhances the PCPAS signal amplitude over that from the bare Hg(HDz)2 film by almost sixfold. The standard calibration graph of PCPAS signal amplitude as a function of Hg2+ concentration is linear in the concentration range of 5-100 μg/ml. A lower detection limit can be achieved by using a laser of higher power tuned to a wavelength closer to the maximum absorptivity of the excited Hg(HDz)2 complex. A study, conducted to monitor the change in PCPAS signal amplitude obtained for the same sample over an extended storage period of 19 days, demonstrates that the PVC protects the integrity of the solid Hg(HDz)2 sample. Hence, it is potentially feasible to collect environmental samples from a remote area for analysis at a later date

Preliminary performance assessment of biotoxin detection for UWS applications using a MicroChemLab device.

In a multiyear research agreement with Tenix Investments Pty. Ltd., Sandia has been developing field deployable technologies for detection of biotoxins in water supply systems. The unattended water sensor or UWS employs microfluidic chip based gel electrophoresis for monitoring biological analytes in a small integrated sensor platform. This instrument collects, prepares, and analyzes water samples in an automated manner. Sample analysis is done using the {mu}ChemLab{trademark} analysis module. This report uses analysis results of two datasets collected using the UWS to estimate performance of the device. The first dataset is made up of samples containing ricin at varying concentrations and is used for assessing instrument response and detection probability. The second dataset is comprised of analyses of water samples collected at a water utility which are used to assess the false positive probability. The analyses of the two sets are used to estimate the Receiver Operating Characteristic or ROC curves for the device at one set of operational and detection algorithm parameters. For these parameters and based on a statistical estimate, the ricin probability of detection is about 0.9 at a concentration of 5 nM for a false positive probability of 1 x 10{sup -6}

Rapid onsite assessment of spore viability.

This one year LDRD addresses problems of threat assessment and restoration of facilities following a bioterror incident like the incident that closed down mail facilities in late 2001. Facilities that are contaminated with pathogenic spores such as B. anthracis spores must be shut down while they are treated with a sporicidal agent and the effectiveness of the treatment is ascertained. This process involves measuring the viability of spore test strips, laid out in a grid throughout the facility; the CDC accepted methodologies require transporting the samples to a laboratory and carrying out a 48 hr outgrowth experiment. We proposed developing a technique that will ultimately lead to a fieldable microfluidic device that can rapidly assess (ideally less than 30 min) spore viability and effectiveness of sporicidal treatment, returning facilities to use in hours not days. The proposed method will determine viability of spores by detecting early protein synthesis after chemical germination. During this year, we established the feasibility of this approach and gathered preliminary results that should fuel a future more comprehensive effort. Such a proposal is currently under review with the NIH. Proteomic signatures of Bacillus spores and vegetative cells were assessed by both slab gel electrophoresis as well as microchip based gel electrophoresis employing sensitive laser-induced fluorescence detection. The conditions for germination using a number of chemical germinants were evaluated and optimized and the time course of protein synthesis was ascertained. Microseparations were carried out using both viable spores and spores inactivated by two different methods. A select number of the early synthesis proteins were digested into peptides for analysis by mass spectrometry

Small acid soluble proteins for rapid spore identification.

This one year LDRD addressed the problem of rapid characterization of bacterial spores such as those from the genus Bacillus, the group that contains pathogenic spores such as B. anthracis. In this effort we addressed the feasibility of using a proteomics based approach to spore characterization using a subset of conserved spore proteins known as the small acid soluble proteins or SASPs. We proposed developing techniques that built on our previous expertise in microseparations to rapidly characterize or identify spores. An alternative SASP extraction method was developed that was amenable to both the subsequent fluorescent labeling required for laser-induced fluorescence detection and the low ionic strength requirements for isoelectric focusing. For the microseparations, both capillary isoelectric focusing and chip gel electrophoresis were employed. A variety of methods were evaluated to improve the molecular weight resolution for the SASPs, which are in a molecular weight range that is not well resolved by the current methods. Isoelectric focusing was optimized and employed to resolve the SASPs using UV absorbance detection. Proteomic signatures of native wild type Bacillus spores and clones genetically engineered to produce altered SASP patterns were assessed by slab gel electrophoresis, capillary isoelectric focusing with absorbance detection as well as microchip based gel electrophoresis employing sensitive laser-induced fluorescence detection

Surface plasmon resonance sensors using molecularly imprinted polymers for sorbent assay of theophylline, caffeine, and xanthine

A sensory system based on the optical phenomenon of surface plasmon resonance (SPR), which employs either photothermal deflection spectroscopy (PDS) or a photodiode array (PDA) for detection, was developed to use molecularly imprinted (MI) polymethacrylic acid -ethylene glycol dimethacrylates (PMAA-EDMA) as the sensing element. The MI polymers were first processed by Soxhlet extraction to remove the print molecules (theophylline, caffeine, and xanthine), yielding the specific anti-polymers. Each anti-polymer was layered over a silver film to serve as the analysis surface for the molecularly imprinted sorbent assay (MIA) of one target drugs. This surface was exposed for 60 min to an aqueous standard drug solution, dried in air, and the uptake of the print molecule into the anti- polymer was monitored by shifts in the SPR angle θ(r) (and hence the SPR- PDS signal measured at constant θ). The linear dynamic range of the MIA was found to extend up to 6 mg/mL, with a concentration detection limit estimated at 0.4 mg/mL for theophylline in aqueous solution. A cross-reactivity study of the anti-theophylline and anti-caffein

Microbial agent detection using near-IR electrophoretic and spectral signatures (MADNESS) for rapid identification in detect-to-warn applications.

Rapid identification of aerosolized biological agents following an alarm by particle triggering systems is needed to enable response actions that save lives and protect assets. Rapid identifiers must achieve species level specificity, as this is required to distinguish disease-causing organisms (e.g., Bacillus anthracis) from benign neighbors (e.g., Bacillus subtilis). We have developed a rapid (1-5 minute), novel identification methodology that sorts intact organisms from each other and particulates using capillary electrophoresis (CE), and detects using near-infrared (NIR) absorbance and scattering. We have successfully demonstrated CE resolution of Bacillus spores and vegetative bacteria at the species level. To achieve sufficient sensitivity for detection needs ({approx}10{sup 4} cfu/mL for bacteria), we have developed fiber-coupled cavity-enhanced absorbance techniques. Using this method, we have demonstrated {approx}two orders of magnitude greater sensitivity than published results for absorbing dyes, and single particle (spore) detection through primarily scattering effects. Results of the integrated CE-NIR system for spore detection are presented