Trimethylamine N-oxide dihydrate is a gut microbe-dependent metabolite of dietary choline and other trimethylamine-containing nutrients. Trimethylamine N-oxide dihydrate induces inflammation by activating ROS/NLRP3 inflammasome. Trimethylamine N-oxide dihydrate also accelerates fibroblast differentiation and induces cardiac fibrosis by activating the TGF-β/smad2 signaling pathway.

Physicochemical Properties

Molecular Formula	C3H13NO3
Molecular Weight	111.14
Exact Mass	111.089
CAS #	62637-93-8
Related CAS #	Trimethylamine N-oxide;1184-78-7;Trimethylamine N-oxide-d9;1161070-49-0;Trimethylamine-N-oxide-13C3
PubChem CID	198430
Appearance	White to off-white solid powder
Density	1.157 g/cm3
Melting Point	95-99 °C(lit.)
Flash Point	95 °C
LogP	0.082
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	0
Heavy Atom Count	7
Complexity	28.4
Defined Atom Stereocenter Count	0
InChi Key	PGFPZGKEDZGJQZ-UHFFFAOYSA-N
InChi Code	InChI=1S/C3H9NO.2H2O/c1-4(2,3)5;;/h1-3H3;2*1H2
Chemical Name	N,N-dimethylmethanamine oxide;dihydrate
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)

Biological Activity

Targets	Human Endogenous Metabolite NLRP3 Microbial Metabolite
ln Vitro	When fibroblasts were treated with trimethylamine N-oxide (TMAO) dihydrate in vitro, their size and migration increased in comparison to untreated fibroblasts. Trimethylamine N-oxide dihydrate promotes the phosphorylation of Smad2 and upregulates the expression of collagen I and α-SMA by increasing the expression of TGF-β receptor I. After treating newborn mouse fibroblasts with trimethylamine N-oxide dihydrate, there is a decrease in TGF-betaRI ubiquitination. Additionally, smurf2 expression is inhibited by trimethylamine N-oxide dihydrate[2]. Many marine animals have tissues that contain trimethylamine N-oxide, which is resistant to the negative effects of hydrostatic pressure, high urea, temperature, and salt [3].
ln Vivo	Cardiovascular illness is brought on by dimethylamine N-oxide (TMAO) dihydrate, which increases inflammatory reactions. Three different diets—high choline, standard, or 3-dimethyl-1-butanol (DMB)—were given to C57BL/6 mice. In mice given choline, levels of trimethylamine N-oxide dihydrate and choline were elevated. When HFpEF mice are fed a high-choline diet, heart failure considerably worsens left ventricular hypertrophy, pulmonary congestion, and diastolic dysfunction in comparison to mice fed a control diet. When compared to animals given a control diet, HFpEF mice on a high-choline diet had noticeably higher levels of cardiac fibrosis and inflammation [1].
References	[1]. High-choline Diet Exacerbates Cardiac Dysfunction, Fibrosis, and Inflammation in a Mouse Model of Heart Failure With Preserved Ejection Fraction. J Card Fail. 2020 May 14;S1071-9164(19)31802-0. [2]. Gut Microbe-Derived Metabolite Trimethylamine N-oxide Accelerates Fibroblast-Myofibroblast Differentiation and Induces Cardiac Fibrosis. J Mol Cell Cardiol. 2019 Sep;134:119-130. [3]. Trimethylamine N-Oxide: The Good, the Bad and the Unknown. Toxins (Basel). 2016 Nov 8;8(11):326.

Solubility Data

Solubility (In Vitro)

H2O: 100 mg/mL (899.77 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (22.49 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (22.49 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (22.49 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: 120 mg/mL (1079.72 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	8.9977 mL	44.9883 mL	89.9766 mL
	5 mM	1.7995 mL	8.9977 mL	17.9953 mL
	10 mM	0.8998 mL	4.4988 mL	8.9977 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.