| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
No direct pharmacological target; Hexacosanoic acid-d4-1 is a deuterated internal standard. The unlabeled hexacosanoic acid (C26:0) is a very long-chain fatty acid (VLCFA) that is metabolized by peroxisomal beta-oxidation. It is not a drug target per se, but its abnormal accumulation in the blood and tissues is diagnostic for peroxisomal biogenesis disorders such as X-linked adrenoleukodystrophy (X-ALD), adrenomyeloneuropathy (AMN), Zellweger syndrome, and other peroxisomal disorders. Hexacosanoic acid levels are inversely correlated with the activity of peroxisomal beta-oxidation enzymes (ACOX1, ABCD1). Elevated hexacosanoic acid is associated with increased oxidative stress, inflammation, and myelin instability. Thus, it serves as an important disease biomarker.
|
|---|---|
| ln Vitro |
As a deuterated internal standard, Hexacosanoic acid-d4-1 is not biologically active. The unlabeled hexacosanoic acid (C26:0) is a very long-chain fatty acid involved in lipid metabolism, myelin synthesis, and membrane structure. In patients with X-linked adrenoleukodystrophy (X-ALD) and other peroxisomal disorders, impaired peroxisomal beta-oxidation leads to the accumulation of hexacosanoic acid and other VLCFAs in plasma, red blood cells, and tissues (especially the white matter of the brain, adrenal cortex, and testicular Leydig cells). Elevated hexacosanoic acid levels correlate with disease severity and are used as a diagnostic biomarker. In in vitro studies, hexacosanoic acid (10-100 uM) induces oxidative stress, activates NF-kappaB, and promotes the release of pro-inflammatory cytokines (TNF-alpha, IL-6, IL-1beta) from microglial cells, astrocytes, and macrophages. It may also disrupt myelin stability by integrating into the myelin sheath and altering membrane fluidity. In cultured adrenal cortical cells, hexacosanoic acid impairs steroidogenesis by inhibiting StAR protein expression.
|
| ln Vivo |
No direct in vivo activity for the deuterated compound. The unlabeled hexacosanoic acid accumulates in patients with X-ALD and AMN, leading to neurological deterioration (adrenomyeloneuropathy: spastic paraparesis, sphincter disturbances, adrenal insufficiency) and cerebral demyelination. In animal models (e.g., Abcd1 knockout mice, the mouse model of X-ALD), dietary supplementation with hexacosanoic acid (25-100 mg/kg/day, PO) for 3-6 months increases plasma VLCFA levels, exacerbates oxidative stress, and worsens neurological deficits. Treatment with Lorenzo′s oil (glyceryl trierucate and glyceryl trioleate) or gene therapy aims to normalize hexacosanoic acid levels. Thus, hexacosanoic acid is a disease biomarker and a potential therapeutic target. Hexacosanoic acid-d4-1 is not administered for therapeutic effect but is used as an internal standard to quantify hexacosanoic acid levels in diagnostic and research settings.
|
| Enzyme Assay |
No specific enzyme/receptor binding protocol. For use as an internal standard for the quantification of hexacosanoic acid in plasma, dried blood spots (DBS), or tissue homogenates by GC-MS or LC-MS: (1) Prepare a stock solution of Hexacosanoic acid-d4-1 in hexane, chloroform, or methanol at 1 mg/mL. (2) Prepare calibration standards by spiking known concentrations of unlabeled hexacosanoic acid (analyte, 0.1-100 ug/mL) with a fixed concentration of the deuterated internal standard (e.g., 10 ug/mL) into blank biological matrix (plasma, serum, dried blood spot extracts, or tissue homogenates). (3) For extraction: add 500 uL of chloroform:methanol (2:1) containing internal standard to 100 uL of plasma or 50 uL of DBS extract. (4) Vortex, add 200 uL of 0.9% NaCl, centrifuge at 3000 rpm for 10 min. (5) Collect the lower organic phase, evaporate under nitrogen. (6) For derivatization (GC-MS): add 100 uL of 3 M HCl in methanol to the residue, heat at 65degC for 60 min to form fatty acid methyl esters (FAMEs). (7) Extract FAMEs with hexane, evaporate, reconstitute in hexane. (8) For GC-MS analysis: use a DB-5MS or DB-23 capillary column (30 m × 0.25 mm, 0.25 um), splitless injection, temperature gradient (50degC to 320degC). Detect by EI-MS in selected ion monitoring (SIM) mode: For unlabeled hexacosanoic acid methyl ester (C26:0 methyl ester): m/z 382 (M+), 409 (M+1)?, base ion m/z 74. For Hexacosanoic acid-d4-1 methyl ester: m/z 386 (M+, due to 4 deuteriums), shift of +4 Da. (9) For LC-MS: after extraction, reconstitute in 2-propanol:acetonitrile:water (1:1:1) and separate on a C8 or C18 reverse-phase column. Detect by ESI-MS in negative ion mode: MRM transitions: unlabeled hexacosanoic acid: m/z 395 → 395 (or 395 → ?); Hexacosanoic acid-d4-1: m/z 399 → 399. Use the analyte/internal standard peak area ratio to calculate concentration.
|
| Cell Assay |
(1) For in vitro studies using unlabeled hexacosanoic acid: seed microglial cells (BV-2) or macrophages (RAW 264.7) in 96-well plates at 1 × 10⁵ cells/well. (2) Treat cells with hexacosanoic acid (10-200 uM) conjugated to bovine serum albumin (BSA) at a 1:1 molar ratio to improve solubility. (3) For toxicity: after 24-48 h, add MTT or CCK-8 reagent; measure viability. Hexacosanoic acid concentrations >50 uM may induce cytotoxicity (IC50 ~ 50-100 uM). (4) For oxidative stress: treat cells with hexacosanoic acid (25-100 uM) for 6-24 h, measure intracellular ROS using DCFH-DA (10 uM, 30 min), fluorescence at 485/535 nm. (5) For inflammation: treat BV-2 cells with hexacosanoic acid (25-100 uM) for 6-24 h, collect supernatant, measure TNF-alpha, IL-6, IL-1beta by ELISA. (6) For NF-kappaB activation: treat cells with hexacosanoic acid (25-100 uM) for 0-6 h, lyse, perform Western blot for phospho-p65, total p65, IkappaBalpha. (7) For apoptosis: treat cells for 24-48 h, stain with Annexin V-FITC/PI, analyze by flow cytometry. (8) For metabolic studies: treat cells with hexacosanoic acid-13C (or unlabeled) and measure beta-oxidation products by LC-MS.
|
| Animal Protocol |
(1) For X-ALD mouse model studies: use 8-12 week old male Abcd1 knockout (Abcd1-/ʸ) mice (the murine model of X-ALD, backcrossed to C57BL/6J). (2) Collect blood from the tail vein or by cardiac puncture into EDTA tubes. (3) For dried blood spots (DBS): spot 50-100 uL of whole blood onto Whatman 903 or 903 cards, air dry for 2 h, store at 4degC in a sealed bag with desiccant. (4) Extract hexacosanoic acid from plasma or DBS as described in field 5. Add a fixed concentration of Hexacosanoic acid-d4-1 as an internal standard to each sample. (5) Analyze by GC-MS or LC-MS/MS as described. (6) Calculate plasma hexacosanoic acid levels (expressed in ug/mL or nmol/mL). In X-ALD patients and Abcd1 knockout mice, plasma hexacosanoic acid levels are elevated (2-5× higher than in controls). (7) For diet or drug intervention studies: administer Lorenzo′s oil (40% glyceryl trioleate + 40% glyceryl trierucate, 2-4 mL/kg/day, PO) or statins (atorvastatin, 10-20 mg/kg/day, PO) for 3-6 months; collect blood at baseline and at months 1, 2, 3, 6. (8) Quantify the reduction in plasma hexacosanoic acid levels and correlate with clinical outcomes. (9) For pharmacokinetic studies: administer hexacosanoic acid (50-200 mg/kg, PO) to wild-type mice, collect blood at 0, 2, 4, 6, 8, 12, 24 h, extract, add internal standard, analyze by GC-MS to determine the PK of hexacosanoic acid. (10) For tissue distribution: at endpoint, collect liver, brain, adrenal glands, and testis, homogenize, extract fatty acids, and measure hexacosanoic acid levels using the deuterated internal standard.
|
| ADME/Pharmacokinetics |
Standard formulation for Hexacosanoic acid-d4-1 as an internal standard: prepare a stock solution in hexane, chloroform, or methanol at 0.5-1 mg/mL. Store at -20degC protected from light. For GC-MS analysis of fatty acid methyl esters (FAMEs), prepare a working internal standard solution in hexane or methanol at 10-50 ug/mL. For LC-MS analysis, reconstitute in 2-propanol:acetonitrile:water (1:1:1) or methanol:isopropanol (1:1). Storage: powder stable at -20degC for 3 years, protect from light and moisture; stock solutions in organic solvents stored at -20degC for 6-12 months. Hexacosanoic acid is a saturated, very long-chain fatty acid with very low solubility in water (<0.001 mg/mL) and is soluble in organic solvents (ethanol, chloroform, hexane, DMSO at 10-50 mg/mL). For in vitro studies with unlabeled hexacosanoic acid, conjugate to fatty acid-free BSA (1:1 molar ratio) in PBS or cell culture medium; heat at 37degC for 30 min with sonication to facilitate solubilization. PK of hexacosanoic acid in humans: fasting plasma levels are normally very low (0.1-1.0 ug/mL). In X-ALD patients, levels are elevated (2-10 ug/mL). After oral administration (e.g., in Lorenzo′s oil), plasma hexacosanoic acid levels decline slowly over months due to the reduction of endogenous synthesis and increased beta-oxidation. Half-life in plasma is estimated to be weeks to months in humans (as it is stored in tissues). In rodents, t1/2 of exogenous hexacosanoic acid after IV injection is ~ 2-6 days.
|
| Toxicity/Toxicokinetics |
Hexacosanoic acid is an endogenous very long-chain fatty acid (C26:0) and is not considered acutely toxic at physiological concentrations. However, elevated levels in peroxisomal disorders are neurotoxic and adrenotoxic. In vitro: CCK-8 assay on BV-2 microglial cells with hexacosanoic acid (1-200 uM, 48 h) shows IC50 ~ 50-100 uM. At concentrations >100 uM, significant cytotoxicity, ROS production, and apoptosis are observed. In vivo: acute oral toxicity of hexacosanoic acid in rodents: LD50 > 5000 mg/kg. Chronic exposure (months to years) to elevated hexacosanoic acid levels in X-ALD patients causes progressive neurological disability, adrenal insufficiency, and death. The deuterated internal standard Hexacosanoic acid-d4-1 is used at trace levels (ug/mL) and is non-toxic. The compound is for research use only, not for human therapeutic use. Use standard laboratory safety precautions: gloves, lab coat, eye protection. Avoid inhalation of dust.
|
| References | |
| Additional Infomation |
Hexacosanoic acid-d4-1 is a stable isotope-labeled (deuterium) form of hexacosanoic acid (C26:0, cerotic acid), a very long-chain saturated fatty acid. It is used as an internal standard for the accurate quantification of hexacosanoic acid in plasma, serum, dried blood spots (DBS), and tissue samples by GC-MS or LC-MS in clinical diagnostics and research. Elevated hexacosanoic acid and other very long-chain fatty acids (VLCFAs) are diagnostic biomarkers for X-linked adrenoleukodystrophy (X-ALD), adrenomyeloneuropathy (AMN), Zellweger syndrome, and other peroxisomal disorders. In X-ALD, the ABCD1 gene mutation impairs peroxisomal beta-oxidation, leading to VLCFA accumulation in plasma and tissues, which is used for newborn screening and disease monitoring. Hexacosanoic acid-d4-1 is not a drug and is not FDA-approved; it is strictly for research and diagnostic use (as a reference standard). This product is intended for laboratory research purposes only and is not for human therapeutic or diagnostic use without appropriate regulatory authorization.
|
| Molecular Formula |
C26H48D4O2
|
|---|---|
| Molecular Weight |
400.71
|
| Exact Mass |
400.421
|
| CAS # |
1194984-85-4
|
| Related CAS # |
Hexacosanoic acid;506-46-7
|
| PubChem CID |
91555690
|
| Appearance |
White to off-white solid powder
|
| Density |
0.9±0.1 g/cm3
|
| Boiling Point |
418.7±8.0 °C at 760 mmHg
|
| Flash Point |
187.6±13.3 °C
|
| Vapour Pressure |
0.0±1.0 mmHg at 25°C
|
| Index of Refraction |
1.461
|
| LogP |
12.47
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
2
|
| Rotatable Bond Count |
24
|
| Heavy Atom Count |
28
|
| Complexity |
301
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C(CCCCCCCC(=O)O)CCC([H])([H])C([H])([H])CCCCCCCCCCCCC
|
| InChi Key |
XMHIUKTWLZUKEX-QZPARXMSSA-N
|
| InChi Code |
InChI=1S/C26H52O2/c1-2-3-4-5-6-7-8-9-10-11-12-13-14-15-16-17-18-19-20-21-22-23-24-25-26(27)28/h2-25H2,1H3,(H,27,28)/i14D2,15D2
|
| Chemical Name |
12,12,13,13-tetradeuteriohexacosanoic acid
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
THF :~10 mg/mL (~24.96 mM )
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4956 mL | 12.4779 mL | 24.9557 mL | |
| 5 mM | 0.4991 mL | 2.4956 mL | 4.9911 mL | |
| 10 mM | 0.2496 mL | 1.2478 mL | 2.4956 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
