Simulations of Test Reduction Using Pooled Heavy Metals Analysis in Cannabis.

BACKGROUND: Cannabis species have a propensity to bioaccumulate toxic heavy metals from their growth media. Increased testing for these metals is required to improve the safety of the legal medical and recreational cannabis industries. However, the current methods used for mandated heavy metals tests are not efficient for a large framework. As a result, there is limited testing capacity, high testing costs, and long wait times for results across North America. OBJECTIVE: This study aimed to demonstrate that pooling strategies can be used to increase the throughput in cannabis testing labs and reduce some of the strain on the industry. METHODS: This paper presents an algorithm to simulate different pooling strategies. The algorithm was applied to real world data sets collected from Washington and California state testing labs. RESULTS: Using a single pooling method, a pool size of three samples on average resulted in a 23.8% reduction in tests required for 100 samples for the Washington lab. For the California lab, pooling four samples on average resulted in a 54.1% reduction in tests required for 100 samples. CONCLUSION: The algorithms generated from the Washington and California lab data demonstrated that pooled testing strategies can be developed on a case-by-case method to reduce the time, effort, and costs associated with heavy metals tests. HIGHLIGHTS: The benefits of pooled testing will vary depending on the region and rate of contamination seen in each testing lab. Overall, our results demonstrate pooled testing has the potential to reduce the fiscal costs of testing through increased efficiency, allowing increased testing, leading to greater safety.

Strategies for Nonpolar Aerosol Collection and Heavy Metals Analysis of Inhaled Cannabis Products.

The rapid growth of inhalable cannabis concentrates raises questions about the safety of acute and chronic exposure to these aerosol mixtures. Due to the nonpolar nature of the aerosol mixture created from cannabis vapor cartridges, traditional aqueous-based capture methods used in e-cigarette or tobacco cigarette studies for analysis of metals are insufficient. Moreover, hydrophobic cannabis concentrates are not miscible with dilute aqueous acids and therefore not ideal for metal spiking unlike electronic nicotine delivery systems. This study describes a method of spiking nonaqueous matrices with aqueous metals standards to investigate aerosolization and recovery of the metals. It also compares various methods for nonpolar aerosol capture and subsequent analysis of 10 metals (As, Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb, and Sn) in two model cannabis matrices, flower and concentrate. Spiked cannabis concentrates were vaped in commercially available cartridges, and their aerosol mixtures were investigated for recovery of heavy metals via ICP-MS. Spiked flower samples were also combusted to compare collection rates of the 10 metals. Results show that not all metals that are present in the concentrate or flower can be fully recovered in the aerosol capture processes at standard voltage settings or combustion temperatures. These studies also demonstrate the importance of a nonpolar solvent as part of the aerosol collection to increase the recovery of some metals. The high concentration of some metals seen in the concentrate suggests that the devices themselves are potential routes of exposure. The ICP-MS analysis method was further validated by evaluating different parameters including linearity, matrix effect, limit of detection, limit of quantitation, and repeatability.

Discrete arrays of liquid-crystal-supported proteolipid monolayers as phantom cell surfaces.

The phospholipid bilayers of living cell membranes exist almost universally in a liquid state. This enables motion and spatial reorganization of membrane components on multiple length scales, which is an essential feature of many biological processes. There is great interest in the development of molecularly defined interfaces between synthetic materials and living cells. To this end, there is a need for solid substrate materials that can be derivatized with fluid, membrane-like interfaces. Herein, we describe array fabrication of discrete liquid-crystal areas supporting phospholipid monolayer membranes, and characterize the interactions with several different membrane surface proteins [avidin series, cholera toxin, green fluorescent protein (GFP), intercellular adhesion molecule (ICAM) and major histocompatibility complex (MHC)]. Three different linkage strategies (biotin, nickel chelating lipids complexing with histidine, and the choleratoxin binding unit (CTB) associating with G(M1) are evaluated. Additionally, experiments with live immunological T cells forming active synapses at the interface exhibit the specific nature of the surface.

Synthetic analogues of glycosylphosphatidylinositol-anchored proteins and their behavior in supported lipid bilayers.

Positioned at the C-terminus of many eukaryotic proteins, the glycosylphosphatidylinositol (GPI) anchor is a posttranslational modification that anchors the modified proteins in the outer leaflet of the plasma membrane. GPI-anchored proteins play vital roles in signal transduction, the vertebrate immune response, and the pathobiology of trypanosomal parasites. While many GPI-anchored proteins have been characterized, the biological functions of the GPI anchor have yet to be elucidated at a molecular level. We synthesized a series of GPI-protein analogues bearing modified anchor structures that were designed to dissect the contribution of various glycan components to the GPI-protein's membrane behavior. These anchor analogues were similar in length to native GPI anchors and included mimics of the native structure's three domains. A combination of expressed protein ligation and native chemical ligation was used to attach these analogues to the green fluorescent protein (GFP). These modified GFPs were incorporated in supported lipid bilayers, and their mobilities were analyzed using fluorescence correlation spectroscopy. The data from these experiments suggest that the GPI anchor is more than a simple membrane-anchoring device; it also may prevent transient interactions between the attached protein and the underlying lipid bilayer, thereby permitting rapid diffusion in the bilayer. The ability to generate chemically defined analogues of GPI-anchored proteins is an important step toward elucidating the molecular functions of this interesting post-translational modification.

Colorimetric bio-barcode amplification assay for cytokines.

The bio-barcode amplification assay has become a powerful tool in detecting tens to hundreds of biological targets such as proteins and nucleic acids in the entire sample. However, current bio-barcode detection schemes still require many experimental steps including microarrayer-based immobilization of oligonucleotides on a glass chip, silver enhancement of immobilized gold nanoparticles on a chip, and light-scattering measurement. Here, we report a colorimetric bio-barcode method that minimizes the above requirements while detecting 30 aM concentrations of cytokines (approximately 3 orders of magnitude more sensitive than conventional nonenzymatic cytokine detection assays). The assay is based on porous microparticles, which enable loading of a large number of barcode DNA per particle, and gold nanoparticle-based colorimetric barcode detection method.