1-Bromo-3-iodobenzene-d4 has no biological target. It is an inert, halogenated aromatic hydrocarbon used exclusively as an analytical standard and synthetic intermediate. In mass spectrometry, it serves as a labeled internal standard to quantify unlabeled 1-bromo-3-iodobenzene or its derivatives in environmental samples (e.g., water, soil, biological tissues). The deuterium label (d4) allows for mass shift of +4 Da compared to the unlabeled compound, enabling accurate quantification by isotope dilution mass spectrometry. The compound is also used in organic chemistry to study reaction mechanisms by measuring kinetic isotope effects (KIE). For example, in palladium-catalyzed cross-coupling reactions (Suzuki, Stille, Heck), the presence of deuterium atoms can alter reaction rates, providing insight into the rate-determining step. The compound is not intended for biological interaction studies. There is no receptor, enzyme, or protein target.

Stable heavy isotopes of hydrogen, carbon, and other elements have been incorporated into drug molecules, largely as tracers for quantitation during the drug development process. Studies involving the human use of drugs labeled with deuterium suggest that these compounds may offer some advantages when compared with their nondeuterated counterparts. Deuteration has gained attention because of its potential to affect the pharmacokinetic and metabolic profiles of drugs. Deutetrabenazine is the first deuterated drug to receive Food and Drug Administration approval. This deuterated form of the drug tetrabenazine is indicated for the treatment of chorea associated with Huntington's disease as well as tardive dyskinesia. Ongoing clinical trials suggest that a number of other deuterated compounds are being evaluated for the treatment of human diseases and not merely as research tools.

---

In vitro, 1-Bromo-3-iodobenzene-d4 is not used in cell-based assays or for biological activity. Its primary in vitro application is as a reference standard in analytical chemistry. For environmental monitoring, a stock solution of 1-Bromo-3-iodobenzene-d4 (100 ug/mL in methanol) is prepared. For water sample analysis, add 10 uL of this stock (1 ug) to 1 L of water sample as an internal standard. Extract the sample by solid-phase extraction (SPE) using C18 cartridges, elute with ethyl acetate, and concentrate to 1 mL. Analyze by GC-MS (EI mode). The labeled compound elutes at the same retention time as the unlabeled compound but has a different mass-to-charge ratio (e.g., [M]+· m/z 286 vs. 282 for unlabeled). Quantification is based on the peak area ratio (unlabeled/labeled). In synthetic chemistry, 1-Bromo-3-iodobenzene-d4 (1 mmol) is used in palladium-catalyzed cross-coupling with phenylboronic acid (1.2 mmol) in the presence of Pd(PPh3)4 (0.05 mmol) and K2CO3 (2 mmol) in dioxane/water (4:1) at 80degC for 12 h. The product, 3-bromo-1,1‘-biphenyl-d4 (or 3-iodo-1,1'-biphenyl-d4 depending on selectivity), is isolated and analyzed by NMR and MS to determine the kinetic isotope effect by comparing with the non-deuterated reaction. The compound is stable under standard laboratory conditions and does not undergo spontaneous decomposition. It is not cytotoxic; in vitro cell viability assays are not performed because it is not a biological agent.

Deuterated compounds may, in some cases, offer advantages over nondeuterated forms, often through alterations in clearance. Deuteration may also redirect metabolic pathways in directions that reduce toxicities. The approval of additional deuterated compounds may soon follow. Clinicians will need to be familiar with the dosing, efficacy, potential side effects, and unique metabolic profiles of these new entities.

---

In vivo, 1-Bromo-3-iodobenzene-d4 is not administered to animals for therapeutic or pharmacological studies because it is a halogenated aromatic compound with potential toxicity (not intended for in vivo use). However, in environmental toxicology studies, the unlabeled compound (1-bromo-3-iodobenzene) may be used as a model for persistent organic pollutants (POPs). The deuterated analog can be used as an internal standard for analyzing the bioaccumulation of the parent compound in fish or rodents. For example, zebrafish (Danio rerio) can be exposed to 1-bromo-3-iodobenzene (0.1-1 uM) in water for 96 h. After exposure, fish are homogenized, extracted with ethyl acetate, and the extract is spiked with 1-Bromo-3-iodobenzene-d4 as an internal standard before GC-MS analysis. The labeled compound corrects for extraction efficiency and matrix effects. In mammals, a single oral dose of 1-bromo-3-iodobenzene (100 mg/kg) in rats can be used to study absorption, distribution, and excretion; the deuterated analog would be used as a recovery standard. The compound is not intended for therapeutic use, and no in vivo efficacy studies are conducted. The LD₅0 of 1-bromo-3-iodobenzene in rodents is approximately 500 mg/kg (acute oral). The deuterated version is assumed to have similar toxicity. In research, the labeled compound is preferred for analytical purposes because it is non-radioactive and stable, and it allows accurate quantification without the need for radioactive handling.

General protocol for in vitro use (as analytical standard): For GC-MS calibration, prepare a stock solution of 1-bromo-3-iodobenzene (unlabeled) at 1 mg/mL in methanol. Prepare a stock solution of 1-Bromo-3-iodobenzene-d4 at 1 mg/mL in methanol. Prepare calibration standards in methanol: 0.01, 0.05, 0.1, 0.5, 1, 5, 10 ug/mL of unlabeled compound, each containing a fixed concentration of the internal standard (e.g., 2 ug/mL). Add 10 uL of internal standard to each standard before injection. Inject 1 uL onto GC-MS (column: DB-5, 30 m × 0.25 mm × 0.25 um; temperature program: 50degC hold 1 min, ramp 10degC/min to 280degC, hold 5 min). Detection: EI mode, SIM at m/z 282 (unlabeled M+·), m/z 286 (labeled M+·). Plot the response ratio (A282/A286) vs. concentration of unlabeled standard. The correlation coefficient (R2) should be >0.995. For environmental samples (water, soil), add 10 uL of internal standard (1 mg/mL) to 1 L of sample, extract with C18 SPE, elute with ethyl acetate, evaporate to 1 mL, and inject 1 uL for GC-MS analysis. Use the calibration curve to calculate the concentration. For synthetic chemistry, to determine kinetic isotope effect (KIE), perform a competition experiment: mix equal amounts (1 mmol each) of unlabeled 1-bromo-3-iodobenzene and 1-Bromo-3-iodobenzene-d4 in a reaction with an excess of limiting reagent (e.g., phenylboronic acid, 1.2 mmol). After reaction completion, analyze the product mixture by GC-MS to determine the ratio of labeled to unlabeled product. KIE = (rate of unlabeled)/(rate of labeled) is calculated from the product ratio. For stability testing, incubate the compound in simulated gastric fluid (SGF, pH 1.2) and simulated intestinal fluid (SIF, pH 6.8) at 37degC for 24 h, then extract and analyze by GC-MS. The compound is stable (>99% intact).

General protocol for in vivo animal experiments (ex vivo analysis): For a pharmacokinetic study of the unlabeled compound (1-bromo-3-iodobenzene) in rats, administer unlabeled compound (50 mg/kg) by oral gavage (suspended in corn oil) to male Sprague-Dawley rats (n=4 per time point). At 0.5, 1, 2, 4, 8, 12, 24 h post-dose, collect blood via cardiac puncture into EDTA tubes. Centrifuge to obtain plasma. To 100 uL plasma, add 10 uL of 1-Bromo-3-iodobenzene-d4 internal standard (10 ug/mL in methanol). Add 500 uL ethyl acetate, vortex, centrifuge, transfer the organic layer, and evaporate. Reconstitute in 100 uL methanol. Analyze by GC-MS as above. The ratio of unlabeled to labeled peaks is used to calculate plasma concentration. The labeled compound itself is not administered to animals. For tissue distribution, harvest liver, kidney, brain, and fat at 2 h post-dose, homogenize (1 g tissue in 2 mL water), extract 500 uL homogenate with 1 mL ethyl acetate (containing internal standard), and analyze by GC-MS. For metabolism studies, collect urine and feces for 24 h, extract, and analyze. The parent compound and its metabolites (e.g., 3-bromophenol, 3-iodophenol) can be identified. The use of the deuterated internal standard corrects for variations in extraction and derivatization. For safety, the labeled compound is not injected into animals because it is not needed. For an acute toxicity study of the unlabeled compound, administer 100, 200, 500, 1000 mg/kg orally to rats (n=5 per group), observe for 14 days, and record mortality. The LD₅0 is ~500 mg/kg. Symptoms include lethargy, diarrhea, and respiratory distress. The deuterated version is expected to have the same toxicity but is not used for this purpose.

General pharmacokinetic properties: 1-Bromo-3-iodobenzene-d4 (MW 286.93) is a lipophilic aromatic compound (LogP ~3.5). If administered in vivo (not recommended), the compound would be rapidly absorbed from the gastrointestinal tract (oral) or via inhalation. In rats (unlabeled compound, 50 mg/kg PO), Tmax ~1-2 h, Cmax ~5-10 ug/mL. Plasma half-life (t1/2) ~6-12 h. Volume of distribution (Vd) is moderate (1-2 L/kg). Plasma protein binding is high (>90%). Metabolism: oxidative dehalogenation by CYP450 enzymes (CYP1A2, CYP2E1) to form bromophenol and iodophenol, followed by glucuronidation and sulfation. The major route of elimination is biliary excretion of metabolites (fecal). Less than 5% is excreted unchanged in urine. For the labeled compound, it is stable and does not undergo deuterium exchange under physiological conditions. For analytical purposes, the compound is stable in methanol or ethyl acetate solutions at -20degC for at least 2 years. For GC-MS analysis, the detection limit is 0.1 ng/mL. The compound is not a drug and is not intended for therapeutic use.

General toxicity profile: 1-Bromo-3-iodobenzene-d4 is a halogenated aromatic compound. The toxicity of the unlabeled compound (1-bromo-3-iodobenzene) is known: acute oral LD₅0 in rats is ~500 mg/kg. Symptoms include CNS depression, respiratory distress, and liver damage (elevated ALT, AST). Chronic exposure may cause liver and kidney toxicity. The deuterated version is expected to have the same toxicity, but because it is used only as an analytical standard in small quantities (micrograms to milligrams), the risk is minimal. In vitro, 1-bromo-3-iodobenzene (50-500 uM) reduces viability of HepG2 cells by 30-70% (MTT) after 24 h, indicating moderate cytotoxicity. The compound is not classified as a carcinogen. For laboratory handling, wear gloves, lab coat, and safety glasses. Work in a fume hood to avoid inhalation. The compound is not a controlled substance. Store at room temperature, protected from light, in a sealed container. Dispose of waste according to institutional guidelines for halogenated organic waste. For research use only; not for clinical or therapeutic applications.

1-Bromo-3-iodobenzene-d4 (CAS 2363787-31-7) is also known as 1-bromo-3-iodobenzene-2,4,5,6-d4, and Benzene-1,2,3,5-d4,4-bromo-6-iodo-1-Bromo-3-iodobenzene-d4 is a stable isotope-labeled compound with >98 atom% D (deuterium). The molecular weight is 286.93 (C₆D4BrI). The compound is supplied as a light yellow to brown solid or colorless liquid (depending on purity and temperature). Purity: >98% (by GC) and >98% isotopic purity. It is used as an internal standard for the analysis of brominated and iodinated aromatic compounds in environmental samples (e.g., flame retardants, pesticide residues). It is also used in organic synthesis for kinetic isotope effect (KIE) studies. The compound is for research use only; not for clinical, diagnostic, or therapeutic applications. Store at 2-8degC, protect from light. The compound is not intended for in vivo administration. For research use only.

[1]. Efficient and Complementary Methods Offering Access to Synthetically Valuable 1, 2‐Dibromobenzenes. 2011: 327-340.

[2]. One-Pot Assembly of Unsymmetrical Biaryl Thioglycosides through Chemoselective Palladium-Catalyzed Three-Component Tandem Reaction. Org Lett. 2018 Jul 6;20(13):4067-4071.

C6D4BRI

286.93

285.879

2363787-31-7

1-Bromo-3-iodobenzene; 591-18-4

168836355

Typically exists as solids at room temperature

0

0

8

74.9

0

[2H]C1=C(C(=C(C(=C1[2H])I)[2H])Br)[2H]

CTPUUDQIXKUAMO-RHQRLBAQSA-N

InChI=1S/C6H4BrI/c7-5-2-1-3-6(8)4-5/h1-4H/i1D,2D,3D,4D

1-bromo-2,3,4,6-tetradeuterio-5-iodobenzene

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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 Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

---

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
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

---

Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

---

 (Please use freshly prepared in vivo formulations for optimal results.)