Analytical Methodology
The multi-modal assessment workflow combines three sequential stages — PBMC isolation, HRMS chemical analysis, and EIS functional analysis — to deliver a comprehensive picture of both toxin burden and cellular impact.
PBMC Isolation
Sample preparation — the foundation of the entire multi-modal assessment.
Peripheral Blood Mononuclear Cells (PBMCs) — comprising lymphocytes and monocytes — are isolated from a standard blood draw using density gradient centrifugation (e.g., Ficoll-Paque™). These long-lived immune cells accumulate toxins over time and serve as a faithful, non-invasive surrogate for the body's true tissue toxin burden.
Technical Principle
Density gradient centrifugation separates PBMCs from red blood cells, granulocytes, and plasma based on their buoyant density. The resulting PBMC layer is washed, counted, and assessed for viability before downstream analysis.
Key Advantages
Non-invasive: requires only a standard blood draw
Reflects cumulative, long-term intracellular toxin burden
Provides both chemical (HRMS) and functional (EIS) analysis from a single sample
Applicable to humans and animals
High-Resolution Mass Spectrometry
Chemical analysis — identifies and quantifies the specific toxins present within the PBMCs.
HRMS is the analytical gold standard for identifying and quantifying a comprehensive panel of environmental toxins within isolated PBMCs. It provides definitive molecular identification with sub-parts-per-trillion sensitivity, covering the full spectrum of chemical toxin classes in a single analytical run.
Technical Principle
Molecules are ionized and separated by their precise mass-to-charge ratio (m/z). High-resolution instruments (e.g., Orbitrap, Q-TOF) provide mass accuracy better than 5 ppm, enabling unambiguous identification of known and unknown toxins. LC-HRMS is used for polar compounds; GC-HRMS for non-polar and volatile compounds.
Key Advantages
Unmatched sensitivity and specificity for complex chemical mixtures
Simultaneously detects hundreds of toxins across all chemical classes
Inherently quantitative — the gold standard for accurate concentration measurement
Capable of non-targeted screening to detect novel or unexpected toxins
Electrochemical Impedance Spectroscopy
Functional analysis — measures the biological impact and cellular damage caused by the toxin burden.
EIS in a microfluidic device measures the electrical properties of individual PBMCs to assess their functional health and integrity. Changes in intracellular electrical capacity (iEC) serve as a direct, quantitative biomarker of cellular damage caused by toxin exposure.
Technical Principle
PBMCs are introduced into a microfluidic channel with interdigitated micro-electrodes. An alternating electrical current is applied across a frequency range (1 kHz – 1 MHz). The resulting impedance spectrum is analyzed to extract cellular parameters including membrane capacitance, cytoplasmic conductivity, and intracellular electrical capacity (iEC).
Key Advantages
Label-free and non-destructive measurement of live cells
Provides a direct functional readout of cellular damage — not just chemical presence
High throughput: capable of analyzing thousands of cells per minute
Complements HRMS by linking chemical exposure to biological effect
"By integrating the specific chemical data from HRMS with the functional biological data from EIS, this invention provides a holistic, robust, and clinically actionable assessment of a subject's true environmental toxin burden and its pathophysiological effects."
— From the Summary of the Invention