Overview of Relationship Between Lipid and Obesity

Obesity manifests as a pathophysiologic entity amplifying susceptibility to multiorgan pathologies through lipidomic dysregulation. A cardinal manifestation involves dysregulated lipid profiles marked by elevated circulatory lipid species, particularly non-esterified fatty acid moieties, that induce insulin signaling dysfunction across critical metabolic tissues (myocytes, hepatocytes, vascular endothelia). These lipid derivatives establish molecular bridges connecting adiposity, metabolic dyshomeostasis, and vasculopathic processes through pleiotropic mechanisms. Beyond metabolic interference, lipid mediators instigate subclinical inflammatory cascades within musculoskeletal, hepatic, and adipose compartments, potentially accelerating atherogenic endpoints.

At Protheragen, our blood lipid testing service undertakes a meticulous analysis of fatty acids. This thorough examination accurately evaluates the metabolic abnormalities related to obesity. This comprehensive assessment furnishes indispensable data that underpin targets for developing anti-obesity therapeutics. We use the insights gleaned from the metabolic aberration mechanisms uncovered by blood lipid testing in the research of targets for developing anti-obesity therapeutics. Nonetheless, the viability of these identified targets requires rigorous validation in animal models, a process typically undertaken in preclinical studies of anti-obesity therapeutics. Through a battery of assessments encompassing histopathological appraisals, metabolic parameter surveillance, and toxicity evaluations, the effectiveness and safety of prospective drugs are thoroughly ascertained.

Precisely Gauge Lipids for Peak Cardiovascular Well-being

Lipid Extraction

We begin by extracting total lipids from whole blood using a safer solvent system of tert-butyl methyl ether (tert-BME) and methanol. After adding tert-BME and methanol to the blood sample, we vortex the mixture vigorously to ensure thorough lipid dissolution, followed by centrifugation to separate the upper tert-BME layer containing the lipids. This method replaces toxic chloroform, minimizing health risks and environmental impact while maintaining comparable extraction efficiency.

Lipid Separation

We load the tert-BME extract onto a pre-conditioned silica gel solid-phase extraction (SPE) column. Using sequential elution, we isolate cholesterol esters (CE) with methyl acetate in hexane, triacylglycerols (TAG) with methyl acetate in hexane, and glycerophospholipids (GPL) with pure methanol after washing the column with acetone to remove impurities.

Fatty Acid Methyl Ester (FAME) Synthesis

Next, we perform base-catalyzed methanolysis under mild conditions to synthesize FAME. For CE via SPE, we react the lipid with methanolic sodium methacrylate (CH₃ONa) in a hexane/acetone/methanol mixture at room temperature. TAG is rapidly converted to FAME by vortexing with CH₃ONa and acetone, while GPL undergoes complete methanolysis using CH₃ONa in methanol alone. To address residual free fatty acids (FFA) generated during CE methanolysis, we add HCl in methanol and incubate, ensuring complete methylation. Acid-catalyzed methanolysis is reserved for analyzing N-acyl lipids like sphingomyelin.

FAME Purification

We purify the resulting FAME by liquid-liquid extraction with hexane, followed by passing the hexane layer through a silica gel SPE column to remove non-FAME contaminants such as cholesterol, pigments, and unreacted lipids. The purified FAME elutes in 1% methyl acetate/hexane, which we concentrate under vacuum.

Fatty Acid Analysis

Lipid components, including LCFAs, are quantified using advanced analytical techniques like gas chromatography-mass–mass spectrometry (GC-MS). This method separates and identifies fatty acids based on their molecular weight and fragmentation patterns, allowing precise measurement of specific fatty acids.

Workflow

Applications

• Our service can be used to detect abnormal lipid profiles in obese individuals, which are associated with metabolic dysregulation and increased risk of multiorgan pathologies.
• By analyzing lipidomic data, the service provides insights into molecular mechanisms linking obesity, insulin resistance, and inflammation, aiding in the identification of potential therapeutic targets for anti-obesity drugs.
• Our service is useful for supporting preclinical studies, including pharmacodynamic and pharmacokinetic evaluations, to validate the effectiveness and safety of various anti-obesity therapies, such as small molecules, gene therapy, and nanotechnology-based treatments.

Advantages

• We utilize a non-toxic solvent system for lipid extraction, minimizing health risks and environmental impact while maintaining high efficiency.
• Our team integrates advanced techniques for precise lipid profiling, enabling accurate quantification of lipids.
• Our researchers combine lipidomic insights with rigorous in vivo validation (pharmacodynamic/pharmacokinetic studies, toxicity assessments) to accelerate the development of anti-obesity therapies.

Publication Data

Frequently Asked Questions

1. How does blood lipid data guide anti-obesity drug development?

   Lipidomics identifies molecular targets (e.g., hormones, receptors, enzymes) and evaluates drug efficacy in improving lipid dysregulation (e.g., lowering NEFA, modulating LDL subtypes) using animal models.

2. What other diseases can dyslipidemia lead to?
   • Cardiovascular diseases
   • Atherosclerosis
   • Pancreatitis
   • Metabolic syndrome

Protheragen's blood lipid testing service, an integral component of our Obesity Physiology and Biochemistry Analysis Service, specializes in advanced lipidomic profiling to decode obesity-related metabolic dysregulation. Leveraging a chloroform-free solvent system for lipid extraction. By analyzing lipids, the service elucidates the pathophysiological mechanisms driving obesity. If you are interested in our end-to-end solutions—from sample processing to data interpretation, contact us! We help researchers accelerate anti-obesity therapeutic development.

All of our services and products are intended for preclinical research use only and cannot be used to diagnose, treat or manage patients.

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