Coumarin 6 is a fluorescent dye reagent that can be used as a fluorescent probe in microparticle drug delivery systems to conduct in vivo tracking, cellular uptake and transport mechanism studies of active molecule drug delivery systems (λexc=450 nm, λem=505 nm).

Physicochemical Properties

Molecular Formula	C20H18N2O2S
Molecular Weight	350.4341
Exact Mass	350.109
CAS #	38215-36-0
PubChem CID	100334
Appearance	Yellow to orange solid powder
Density	1.311 g/cm3
Boiling Point	570.1ºC at 760 mmHg
Melting Point	208-210 °C(lit.)
Flash Point	298.6ºC
Index of Refraction	1.69
LogP	4.915
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	4
Heavy Atom Count	25
Complexity	534
Defined Atom Stereocenter Count	0
InChi Key	VBVAVBCYMYWNOU-UHFFFAOYSA-N
InChi Code	InChI=1S/C20H18N2O2S/c1-3-22(4-2)14-10-9-13-11-15(20(23)24-17(13)12-14)19-21-16-7-5-6-8-18(16)25-19/h5-12H,3-4H2,1-2H3
Chemical Name	3-(1,3-benzothiazol-2-yl)-7-(diethylamino)chromen-2-one
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 Note: This product requires protection from light (avoid light exposure) during transportation and storage.
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

ln Vitro	HPMC nanoparticles tagged with coumarin 6 [3] (1) Prepare an organic phase solution containing 5 mg/mL by dissolving coumarin 6 in DMF. (2) HBSS-represented dissolved concentration supplemented cellulose (3) Using a MasterFlex L/S pump, add the organic solution to the aqueous solution (volume ratio: 1:100) at a 36 mL/min injection rate (4) Add 9 mL (5) of 2 μg/mL nanoparticles to the MDCKII cell culture dish and shield from light for five, fifteen, thirty, and sixty minutes. (6) To prevent MDCKII cell food nanoparticles, add cold PBS buffer and rinse with HBSS. (7) Examine the MDCKII cells' meal on nanoparticles under a fluorescent microscope.
References	[1]. Preparation of PLGA Nanoparticles Encapsulated with Fluorescent Probe Coumarin-6. bioRxiv, 2019: 614875. [2]. Cellular uptake of coumarin-6 as a model drug loaded in solid lipid nanoparticles. J Physiol Pharmacol. 2011 Feb;62(1):45-53. [3]. Transport Mechanism of Coumarin 6 Nanocrystals with Two Particle Sizes in MDCKII Monolayer and Larval Zebrafish. ACS Appl Mater Interfaces. 2016 May 25;8(20):12620-30.
Additional Infomation	Coumarin 6 is a member of 7-aminocoumarins. It has a role as a fluorochrome. Coumarin 6 has been reported in Ferula fukanensis with data available.

Solubility Data

Solubility (In Vitro)

DMSO : ~5 mg/mL (~14.27 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 (e.g. IP/IV/IM/SC) 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). 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 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). 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.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.8536 mL	14.2682 mL	28.5364 mL
	5 mM	0.5707 mL	2.8536 mL	5.7073 mL
	10 mM	0.2854 mL	1.4268 mL	2.8536 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.