ExCALiBR: Extractor for Chemical Analysis of Lipid Biomarkers in Regolith

Lipids are organic molecules of great interest for future NASA life detection missions. Lipids are essential for life as we know it, and likely also required for putative extraterrestrial organisms. These robust organics are also synthesized abiotically--though with a molecular distribution distinguishable from that characteristic of life--and survive for long periods of geological time. We developed a fluidic system (TRL-4) integrating capabilities and techniques used in traditional lab settings for lipid analysis. This system, the Extractor for Chemical Analysis of Lipid Biomarkers in Regolith (ExCALiBR), will enable future astrobiology surveys by extracting, purifying, and concentrating lipids from approximately 25 cc of regolith. ExCAliBR consists of a comminutor, solvent extractor with sonicator, filter, and concentrator. Goals and Objectives. Our goal is produce a TRL-5 system that has a planetary-payload form-factor to accept unprocessed regolith, mechanically process/pulverize these samples, perform a liquid extraction of lipid biomarkers, and transport lipid extract for downstream mass spectroscopic analysis. Our objectives are to: 1. Improve the ExCALiBR breadboard by reducing the power, mass, and volume of ExCALiBR to planetary instrumentation scale and designing for radiation tolerance. 2. Demonstrate the feasibility of reducing the background contamination signal below that expected from an exobiological sample (to <1 nmol) and characterize total lipid organics carryover between samples. 3 Develop closed-loop control systems and a thermal model as an enabling step toward full automation. 4. Conduct environmental testing including shock and vibration testing of the ExCALiBR system brassboard (at GEVS levels). Additionally, and Mars-like thermal and pressure regime testing to characterize power requirements and system performance. Approach/Methodology. In Year 1, we redesign the ExCALiBR breadboard for spaceflight, leveraging recent development (TRL 2-4) via an STMD Early Career Initiative Award and heritage ARC fluidic architectures from lab-on-a-chip biological and life-detection efforts to minimize volume and power requirements. We perform a lifetime leak test on our solvent handling systems. In Year 2, we improve the design of ExCALiBR components, strengthening the four individual subsystems, and integrate them into a brassboard system. We benchmark instrument performance against traditional laboratory approaches and flight-heritage analytical instrumentation using planetary analog samples (e.g., Atacama soil; Antarctic lake ice). We test decontamination protocols on the integrated system to determine the most effective technique. We Develop closed-loop thermal, ultrasonic-probe, and pressure control systems, as well as thermal model. In Year 3, we conduct mechanical testing (shock and vibe) and thermal/vacuum testing of the fully integrated brassboard to achieve TRL 5. (TRL 6 flightlike environment involves partial gravity testing of the comminutor, which will be accomplished as part of a separately funded effort.) We demonstrate dirt-to-data capability by coupling the brassboard with a laser desorption ionization mass spectrometer. Decontamination technique will be implemented and instrumentation cleanliness validated. We Publish validation results, design, and instrument summary in peer-reviewed journal. Relevance. ExCALiBR is relevant to Objective 1.2 of the 2022 NASA Strategic Plan, which includes "Searching for Life Elsewhere," and states that "NASA will develop tools for detecting life." ExCALiBR will advance the current state of scientific knowledge by characterizing organics in astrobiologically significant regions, responding to the Planetary Decadal Survey 2022-2033 Question 11, "Is there evidence of past or present life in the solar system beyond Earth and how do we detect it?" We are maturing a TRL-4 instrument for inclusion in a future Discovery mission proposal.

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