| Field | Specification |
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| Assay Time | |
| Detection Method | |
| Product Type | |
| Sample Type(s) | Serum, plasma, urine, saliva, milk, cell cultures, food, agriculture etc |
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Overview
For quantitative determination of fatty acid and evaluation of drug effects on its metabolism. The assay uses OD570nm, or FL530/585nm for signal readout. Compatible sample input includes Serum, plasma, urine, saliva, milk, cell cultures, food, agriculture etc. Typical stated assay timing is 30 min.
Key elements and design rationale
- Readout format: OD570nm, or FL530/585nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Serum, plasma, urine, saliva, milk, cell cultures, food, agriculture etc, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 7 µM for interpreting low-signal samples.
- Feature emphasis: Sensitive. Use 10 µL samples. Linear detection range: colorimetric assay 7 – 1000 µM, fluorimetric assay 7 – 100 µM fatty acid.
Additional feature notes highlight Convenient. Room temperature “mix-and-read” procedure can be readily automated for high-throughput assay of thousands of samples per day. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of free fatty acid within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
More details
Fatty acids are aliphatic monocarboxylic acids that are ubiquitously found in animal or vegetable fat, oil, and wax. Fatty Acids play important roles in cellular synthesis, and energy metabolism and are implicated in diverse disorders such as diabetes mellitus, sudden infant death syndrome, and Reye Syndrome. BioAssay Systems method provides a simple, one-step, and high-throughput assay for measuring free fatty acids. In this assay, free fatty acids are enzymatically converted to acyl-CoA and subsequently to H2O2. The resulting H2O2reacts with a specific dye to form a pink-colored product. The optical density at 570nm or fluorescence intensity (530/585 nm) is directly proportional to the free fatty acid concentration in the sample.
Detection method
Colorimetric (OD 570 nm) or Fluorescent (FL 530/585 nm).
Detection limit and analytical sensitivity
Reported detection limit: 7 µM.
Procedures and timing
Stated procedure or timing information: 30 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
Common research applications
- Quantify free fatty acid in serum, plasma, urine by OD570 nm, or FL530/585 nm readout.
- Compare treatment or phenotype groups using matched serum, plasma, urine handling.
- Monitor time-course or pre/post changes in serum, plasma, urine across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
Can you tell me what is the conversion factor for free fatty acids from µM to mEq/L? We are about to measure FFA with your assay (EFFA-100) but most publications in domestic animals are in mEq/L?
1 mEq/L = 1 mmole/L = 1000 µM.
I have just noticed that EDTA can interfere in this assay but, since I used Microvette CB 300 capillary tubes containing Potassium EDTA to collect the blood, I would like to know at what concentration (if known) EDTA is an inhibitor of the reaction.
EDTA in the sample will complex Mg2+ and Ca2+ ions of the assay buffer, necessary cofactors for the enzymes that are used in the assay.
2+
2+
Plasma EDTA tubes usually contain around 5 mM EDTA in the final volume if filled to the draw volume. Much of the EDTA will be complexed with blood Ca2+/Mg2+ ions, and thus not interfere with the assay. The EDTA concentration will be higher if the tubes are only partially filled.
2+
2+
Since the sample to assay buffer ratio is 1:10 and the assay buffer Mg2+ concentration is 1 mM, I would not expect any interference in samples from tubes that have been filled completely. If the collection tube had only been filled partially, I would recommend supplementing the assay working reagent with an additional 1 mM MgCl2.
For laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
Synergism between Corynebacterium and Streptococcus sanguinis reveals new interactions between oral commensals
Treerat, P et al. (2020). Synergism between Corynebacterium and Streptococcus sanguinis reveals new interactions between oral commensals. The ISME Journal, 14(5): 1154-1169. Assay: Fatty acid in C. durum culture supernatant.
Critical role of matrix metalloproteinase 14 in adipose tissue remodeling during obesity
Li, X et al. (2020). Critical role of matrix metalloproteinase 14 in adipose tissue remodeling during obesity. Molecular and Cellular Biology, 40(8). Assay: Fatty acid in rat plasma.
The interaction between cuticle free fatty acids (Ffas) of the cockroaches Blattella germanica and Blatta orientalis and hydrolases produced by the entomopathogenic fungus Conidiobolus coronatus
Kaczmarek, A et al. (2020). The interaction between cuticle free fatty acids (Ffas) of the cockroaches Blattella germanica and Blatta orientalis and hydrolases produced by the entomopathogenic fungus Conidiobolus coronatus. PLoS ONE, 15(7). Assay: Fatty acid in insect cuticle.
Hepatocyte nuclear factor 4a and glucocorticoid receptor coordinately regulate lipid metabolism in mice fed a high-fat-high-sugar diet
Lu, H et al. (2021). Hepatocyte nuclear factor 4a and glucocorticoid receptor coordinately regulate lipid metabolism in mice fed a high-fat-high-sugar diet. BioRxiv, 2021.02.06.427306. Assay: Fatty acid in mouse serum.
Albumin enhances the rate at which coxsackievirus b3 strain 28 converts to a-particles
Carson SD, Cole AJ. (2020). Albumin enhances the rate at which coxsackievirus b3 strain 28 converts to a-particles. Journal of Virology, 94(6). Assay: Fatty acid in bovine serum albumin and culture media.
FABP5 promotes lymph node metastasis in cervical cancer by reprogramming fatty acid metabolism
Zhang, C et al. (2020). FABP5 promotes lymph node metastasis in cervical cancer by reprogramming fatty acid metabolism. Theranostics, 10(15): 6561-6580. Assay: Fatty acid in human cells.
Dietary α-ketoglutarate promotes beige adipogenesis and prevents obesity in middle-aged mice
Tian, Q et al. (2020). Dietary α-ketoglutarate promotes beige adipogenesis and prevents obesity in middle-aged mice. Aging Cell, 19(1), e13059. Assay: Fatty acid in mouse serum.
Sex differences in body composition, metabolism-related hormones, and energy homeostasis during aging in Wistar rats
Quiros Cognuck, S et al. (2020). Sex differences in body composition, metabolism-related hormones, and energy homeostasis during aging in Wistar rats. Physiological Reports, 8(20). Assay: Fatty acid in rat serum.
Variation of body condition and plasma energy substrates with life stage, sex, and season in wild-sampled nurse sharks Ginglymostoma cirratum
Moorhead, SG et al. (2020). Variation of body condition and plasma energy substrates with life stage, sex, and season in wild-sampled nurse sharks Ginglymostoma cirratum. Journal of Fish Biology. Assay: Fatty acid in shark plasma.
Moderate intake of BCAA-rich protein improves glucose homeostasis in high-fat-fed mice
Roquetto, AR et al. (2020). Moderate intake of BCAA-rich protein improves glucose homeostasis in high-fat-fed mice. The Journal of Nutritional Biochemistry, 80, 108332. Assay: Fatty acid in mouse serum.
Grape polyphenols and exercise training have distinct molecular effects on cardiac hypertrophy in a model of obese insulin-resistant rats
Lambert, K et al. (2020). Grape polyphenols and exercise training have distinct molecular effects on cardiac hypertrophy in a model of obese insulin-resistant rats. The Journal of Nutritional Biochemistry, 108522. Assay: Fatty acid in rat plasma.
Novel role of dynamin-related-protein 1 in dynamics of ER-lipid droplets in adipose tissue
Li, X et al. (2020). Novel role of dynamin-related-protein 1 in dynamics of ER-lipid droplets in adipose tissue. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology, 34(6): 8265-8282. Assay: Fatty acid in mouse serum and adipose.
Associations of circulating irisin with fndc5 expression in fat and muscle in type 1 and type 2 diabetic mice
Jiang, S et al. (2021). Associations of circulating irisin with fndc5 expression in fat and muscle in type 1 and type 2 diabetic mice. Biomolecules, 11(2). Assay: Fatty acid in mouse plasma.
Amelioration of obesity-related biomarkers by Lactobacillus sakei CJLS03 in a high-fat diet-induced obese murine model
Ji, Yosep, et al (2019). Amelioration of obesity-related biomarkers by Lactobacillus sakei CJLS03 in a high-fat diet-induced obese murine model. Scientific reports 9.1: 6821. Assay: Free fatty acidin mouse serum.
Saturated Fatty Acid Activates T Cell Inflammation Through a Nicotinamide Nucleotide Transhydrogenase (NNT)-Dependent Mechanism
McCambridge, Grace, et al (2019). Saturated Fatty Acid Activates T Cell Inflammation Through a Nicotinamide Nucleotide Transhydrogenase (NNT)-Dependent Mechanism. Biomolecules 9.2: 79. Assay: Free fatty acidin human plasma.
Buffalo liver transcriptome analysis suggests immune tolerance as its key adaptive mechanism during early postpartum negative energy balance
Singh, Sudhakar, et al (2019). Buffalo liver transcriptome analysis suggests immune tolerance as its key adaptive mechanism during early postpartum negative energy balance. Functional & integrative genomics: 1-15. Assay: Free fatty acidin buffaloe serum.
Gingival periodontal disease (PD) level-butyric acid affects the systemic blood and brain organ: insights into the systemic inflammation of periodontal disease
Cueno, Marni E., and Kuniyasu Ochiai (2018). Gingival periodontal disease (PD) level-butyric acid affects the systemic blood and brain organ: insights into the systemic inflammation of periodontal disease. Frontiers in immunology 9:1158. Assay: Free fatty acidin rat blood cytosol.
IL-25 stimulates M2 macrophage polarization and thereby promotes mitochondrial respiratory capacity and lipolysis in adipose tissues against obesity
Feng, Juan, et al (2018). IL-25 stimulates M2 macrophage polarization and thereby promotes mitochondrial respiratory capacity and lipolysis in adipose tissues against obesity. Cellular & molecular immunology 15.5: 493. Assay: Free fatty acidin mice tissues.
Circulating Adipose Fatty Acid Binding Protein Is a New Link Underlying Obesity-Associated Breast/Mammary Tumor Development
Hao, Jiaqing, et al (2018). Circulating Adipose Fatty Acid Binding Protein Is a New Link Underlying Obesity-Associated Breast/Mammary Tumor Development. Cell metabolism 28.5: 689-705. Assay: Free fatty acidin mice serum.
Combination of nutritional polyphenols supplementation with exercise training counteracts insulin resistance and improves endurance in high-fat diet-induced obese rats
Lambert, Karen, et al (2018). Combination of nutritional polyphenols supplementation with exercise training counteracts insulin resistance and improves endurance in high-fat diet-induced obese rats. Scientific reports 8.1: 2885. Assay: Free fatty acidin rat plasma.
No Additive Effects of Polyphenol Supplementation and Exercise Training on White Adiposity Determinants of High-Fat Diet-Induced Obese Insulin-Resistant rats
Lambert, Karen, et al (2018). No Additive Effects of Polyphenol Supplementation and Exercise Training on White Adiposity Determinants of High-Fat Diet-Induced Obese Insulin-Resistant rats. Oxidative medicine and cellular longevity 2018:7406946. Assay: Free fatty acidin rat plasma.
Mannose Alters Gut Microbiome, Prevents Diet-Induced Obesity, and Improves Host Metabolism
Sharma, Vandana, et al (2018). Mannose Alters Gut Microbiome, Prevents Diet-Induced Obesity, and Improves Host Metabolism. Cell Reports 24.12: 3087-3098. Assay: Free fatty acidin mice serum and feces.
Cuticular fatty acids of Galleria mellonella (Lepidoptera) inhibit fungal enzymatic activities of pathogenic Conidiobolus coronatus
Wronska, Anna Katarzyna, et al (2018). Cuticular fatty acids of Galleria mellonella (Lepidoptera) inhibit fungal enzymatic activities of pathogenic Conidiobolus coronatus. PloS one 13.3: e0192715. Assay: Free fatty acidin moth cuticle.
Energy metabolism in mobile, wild-sampled sharks inferred by plasma lipids
Gallagher, Austin J., et al (2017). Energy metabolism in mobile, wild-sampled sharks inferred by plasma lipids. Conservation physiology 5.1:cox002. Assay: Free fatty acidin sharks plasma.
Moderate alcohol intake induces thermogenic brown/beige adipocyte formation via elevating retinoic acid signaling
Wang, Bo, et al (2017). Moderate alcohol intake induces thermogenic brown/beige adipocyte formation via elevating retinoic acid signaling. The FASEB Journal 31.10: 4612-4622. Assay: Free fatty acidin mice serum.
Retinoic acid induces white adipose tissue browning by increasing adipose vascularity and inducing beige adipogenesis of PDGFRalpha+ adipose progenitors
Wang, Bo, et al (2017). Retinoic acid induces white adipose tissue browning by increasing adipose vascularity and inducing beige adipogenesis of PDGFRalpha+ adipose progenitors. Cell discovery 3: 17036. Assay: Free fatty acidin mice serum.
Intestinal removal of free fatty acids from hosts by Lactobacilli for the treatment of obesity
Chung, Hea-Jong, et al (2016). Intestinal removal of free fatty acids from hosts by Lactobacilli for the treatment of obesity. FEBS Open Bio 6.1: 64-76. Assay: Free fatty acidin human lactobacillus culture.
Body fat mobilization during lactation in high-producing sows fed varied omega-6 to omega-3 fatty acid ratios
Eastwood, Laura, Pascal Leterme, and A. Denise Beaulieu (2016). Body fat mobilization during lactation in high-producing sows fed varied omega-6 to omega-3 fatty acid ratios. Canadian Journal of Animal Science 96.1: 69-78. Assay: Free fatty acidin pig serum.
Pepsin egg white hydrolysate ameliorates obesity-related oxidative stress, inflammation and steatosis in Zucker Fatty rats
Garces-Rimon, M., et al (2016). Pepsin egg white hydrolysate ameliorates obesity-related oxidative stress, inflammation and steatosis in Zucker Fatty rats. PLoS One 11.3: e0151193. Assay: Free fatty acidin rat plasma.
Deletion of apoptosis inhibitor of macrophage (AIM)/CD5L attenuates the inflammatory response and infarct size in acute myocardial infarction
Nishikido, Toshiyuki, et al (2016). Deletion of apoptosis inhibitor of macrophage (AIM)/CD5L attenuates the inflammatory response and infarct size in acute myocardial infarction. Journal of the American Heart Association 5.4: e002863. Assay: Free fatty acidin mice plasma.
Hawthorn fruit extract elevates expression of Nrf2/HO-1 and improves lipid profiles in ovariectomized rats
Yoo, Jeong-Hyun, Yanan Liu, and Hyun-Sook Kim (2016). Hawthorn fruit extract elevates expression of Nrf2/HO-1 and improves lipid profiles in ovariectomized rats. Nutrients 8.5: 283. Assay: Free fatty acidin rat serum.
Hwang I et al (2012) Catalase deficiency accelerates diabetic renal injury through peroxisomal dysfunction. Diabetes 61(
Hwang I et al (2012) Catalase deficiency accelerates diabetic renal injury through peroxisomal dysfunction. Diabetes 61(3):728-738. Assay: Free fatty acidin mouse plasma.
Jelinek D et al (2012) The Niemann-Pick C1 Gene Is Downregulated in Livers of C57BL/6J Mice by Dietary Fatty Acids, but
Jelinek D et al (2012) The Niemann-Pick C1 Gene Is Downregulated in Livers of C57BL/6J Mice by Dietary Fatty Acids, but Not Dietary Cholesterol, through Feedback Inhibition of the SREBP Pathway. J Nutr 142(11):1935-1942. Assay: Free fatty acidin mouse plasma.
Jin Kyubob et al (2012) Dysregulation of hepatic fatty acid metabolism in chronic kidney disease. Nephrol Dial Transplan
Jin Kyubob et al (2012) Dysregulation of hepatic fatty acid metabolism in chronic kidney disease. Nephrol Dial Transplant. Assay: Free fatty acidin rat liver.
Lu Z et al (2012) TLR4 Antagonist Reduces Early-Stage Atherosclerosis in Diabetic Apolipoprotein E-deficient Mice. J End
Lu Z et al (2012) TLR4 Antagonist Reduces Early-Stage Atherosclerosis in Diabetic Apolipoprotein E-deficient Mice. J Endocrinol. Assay: Free fatty acidin mouse serum.
Hepatic and muscular effects of different dietary fat content in VLCAD deficient mice
Primassin, S., S. Tucci, et al. (2011). Hepatic and muscular effects of different dietary fat content in VLCAD deficient mice. Mol Genet Metab 104(4): 546-51. Assay: Free fatty acidin mouse serum.
Antihyperlipidemic and body fat-lowering effects of silk proteins with different fibroin/sericin compositions in mice fed with high fat diet
Seo, C.W., et al. (2011). Antihyperlipidemic and body fat-lowering effects of silk proteins with different fibroin/sericin compositions in mice fed with high fat diet. J Agric Food Chem 59(8):4192-7. Assay: Free fatty acidin mouse plasma.
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