Cannabis Compound Quantification and Calcium Bioassay
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Solution Overview
Problem
Current methods for measuring intracellular calcium levels are time-consuming, and there is a need for a bioassay to determine the concentration of intracellular calcium as a function of cannabinoid or terpene addition to assess the effectiveness of Cannabis-derived compounds in treating various diseases and disorders.
Innovation Solution
A method combining gas chromatography with mass spectrometry for identifying and quantifying cannabinoids and terpenes, along with a bioassay using a microfluidic device to measure intracellular calcium concentration in cells exposed to Cannabis-derived compounds.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If current methods for measuring intracellular calcium levels are used, then measurement accuracy is maintained, but measurement time increases
Solution Approach 1:
The patent replaces traditional mechanical/manual calcium measurement techniques with a fluorescent optical detection system. Fluorescent indicators (such as Fura-2, Fluo-4, or Calcium Green) are introduced into cells, and fluorescent microscopy or flow cytometry is used to detect calcium concentration changes through fluorescence intensity or wavelength shifts, significantly reducing measurement time while maintaining precision
Solution Approach 2:
The patent utilizes changes in fluorescent properties (intensity, wavelength, lifetime) as parameters to indicate calcium concentration. By monitoring these optical parameter changes in real-time, the system achieves rapid and accurate calcium measurement without the time-consuming procedures of traditional methods
2Measurement precision
If gas chromatography with mass spectrometry is used for compound identification, then detection precision is improved, but device complexity increases
Solution Approach 1:
The patent combines gas chromatography (GC) separation technology with mass spectrometry (MS) detection technology into an integrated GC-MS system. The GC component separates complex cannabis compound mixtures into individual components, while the MS component provides precise molecular identification and quantification, achieving high detection precision through functional integration
Solution Approach 2:
The GC-MS system provides multiple functions including separation, identification, and quantification of various cannabis compounds (cannabinoids, terpenes, flavonoids) using a single analytical platform. The system can analyze different compound classes simultaneously, reducing the need for multiple separate analytical instruments
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach allows for fast and efficient detection and quantification of multiple cannabinoids and terpenes, providing real-time measurement of intracellular calcium changes, thus evaluating the pharmacological properties of Cannabis-derived compounds effectively.
Implementation Method 1
separating the cannabinoids by gas chromatography using a capillary column with hydrogen as a carrier gas
Implementation Method 2
detecting the cannabinoids using a mass spectrometer
Implementation Method 3
measuring fluorescence of the cell to determine a background fluorescence (Fmin)
Data Source
AI summary
The disclosure provides a method for analyzing compounds extracted from Cannabis. The method comprises extracting cannabinoids or terpenes from a sample using a C1-C4 alcohol or C5-C8 solvent as an extraction solvent to produce a supernatant, drying the supernatant to produce a dried extract, and dissolving the dried extract in a second C1-C4 alcohol or C5-C8 solvent, separating the cannabinoids or terpenes by gas chromatography using a capillary column with hydrogen as a carrier gas; and detecting the cannabinoids or terpenes using a mass spectrometer. The disclosure also provides a method of determining an effect of one or more Cannabis-derived compounds on intracellular calcium concentration in a cell using a microfluidic device.