Patent Blue V (Acid blue 1) is a new biologic dye that could be utilized as an intraocular dye for retinectomy. Retinectomy involves removal of the translucent inner limiting membrane (ILM). Complete removal can only be achieved by applying appropriate dyes during vitreoretinal surgery. Patent Blue V may be utilized to stain preretinal structures. Spectral analysis shows that Patent Blue V has strong absorption below 450 nm and above 600 nm, showing a blue-green color. Patent Blue V is also used as a lymphangiographic marker for staining during biopsy procedures to remove lymph nodes.

Physicochemical Properties

Molecular Formula	C27H31N2NAO6S2
Molecular Weight	566.66
Exact Mass	566.152
CAS #	129-17-9
PubChem CID	8507
Appearance	VIOLET POWDER Dark bluish-green powder
Melting Point	290ºC
LogP	5.923
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	7
Heavy Atom Count	38
Complexity	1040
Defined Atom Stereocenter Count	0
SMILES	[Na+].CCN(C1=CC=C(C(C2=CC=C(S([O-])(=O)=O)C=C2S([O-])(=O)=O)=C2C=CC(=[N+](CC)CC)C=C2)C=C1)CC |t:22,26|
InChi Key	SJEYSFABYSGQBG-UHFFFAOYSA-M
InChi Code	InChI=1S/C27H32N2O6S2.Na/c1-5-28(6-2)22-13-9-20(10-14-22)27(21-11-15-23(16-12-21)29(7-3)8-4)25-18-17-24(36(30,31)32)19-26(25)37(33,34)35;/h9-19H,5-8H2,1-4H3,(H-,30,31,32,33,34,35);/q;+1/p-1
Chemical Name	sodium;4-[[4-(diethylamino)phenyl]-(4-diethylazaniumylidenecyclohexa-2,5-dien-1-ylidene)methyl]benzene-1,3-disulfonate
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: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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

ADME/Pharmacokinetics	Absorption, Distribution and Excretion Patent blue is selectively absorbed in the lymphatics. When administered orally, patent blue has a very low absorption and limited systemic availability. Patent blue is excreted into the urine and bile.The excretion is more significant in the urine which after the administration of patent blue intravenously can even change to a blue coloration. When administered orally, patent blue is excreted unchanged in feces. This pharmacokinetic parameter has not been studied. This pharmacokinetic property has nos been fully studied. Metabolism / Metabolites In vitro studies have shown that patent blue is not metabolized. Biological Half-Life The elimination half-life is of about 24-48 hours.
Toxicity/Toxicokinetics	Protein Binding Patent blue gets specifically bound to albumin which allows it to travel through the breast lymphatics.
References	[1]. Mennel S, et al. Patent blue: a novel vital dye in vitreoretinal surgery. Ophthalmologica. 2006;220(3):190-3. [2]. Barranger E, et al. Laparoscopic sentinel lymph node procedure using a combination of patent blue and radioisotope in women with cervical carcinoma. Cancer. 2003 Jun 15;97(12):3003-9.
Additional Infomation	Sulfan blue is a dark greenish-black powder. (NTP, 1992) Patent blue is an organic molecular entity. Patent blue is aniline dye and it is one of the most common dyes used. It is a sodium or calcium salt of diethylammonium hydroxide inner salt. It has the chemical designation of (4-(alpha-(p-(diethylamino)phenyl)-2,4-disulfobenzylidene)-2,5-cyclohexadiene-1-ylidene)diethylammonium hydroxide. Patent blue was developed by Guerbet and approved by Health Canada on December 31, 1979. The isomer isosulphan is used in the United States for the same indications than patent blue. Drug Indication Patent blue is used for marking lymphatic vessels and arterial territories as well as for sentinel lymph node prior to biopsy in patients with operable breast cancer and clinically negative lymph nodes in combination with a radiotracer. Patent blue is also used in the textile, paper, agriculture and cosmetic industry. FDA Label Mechanism of Action The specific binding of patent blue allows it to freely travel in the breast lymphatics and allows staging with a less invasive alternative. Patent blue will form a complex with albumin which will be picked up by regional afferent lymphatics to identify sentinel lymph nodes. A sentinel lymph node is the first lymph node in a chain or group of lymph nodes that cancer is most likely to spread to. The determination of the sentinel lymph node allows the physician to stage cancer by observing if cancer has spread to the near lymph nodes. IT IS OF SOME INTEREST THAT SEVERAL DYES RELATED TO PARAROSANILINE, WHICH INCLUDES BLUE VRS, EXHIBIT GENOTOXICITY & THAT THIS ACTIVITY IS CORRELATED WITH THE PRESENCE OF A SUBSTITUTED OR UNSUBSTITUTED AMINO GROUP AT THE R3 POSITION PARA TO THE N+ AMINO GROUP. THUS PATENT BLUE IS INACTIVE. THIS DYE INSTEAD HAS SULFONIC GROUPS IN THIS POSITION WHICH MAY INTERFERE WITH ACTIVATION AT THE N+ ATOM. Therapeutic Uses Dyes EXPTL USE: THE BLUE TEST IS DESIGNED TO REVEAL EDEMA OF TOTAL OR PARTIAL LYMPHATIC ORIGIN. INTOLERANCE TO THE DYE INJECTED IS RARE, THOUGH SOMETIMES SEVERE, & USERS SHOULD BY AWARE OF THE POSSIBILITY. Pharmacodynamics Administration of patent blue has been reported to cause hypersensitivity reactions in approximately 1% of the patients. It also presents a localized blue coloration which has made patent blue a sensitive and specific option for the detection of micrometastatic cancer in lymph nodes.

Solubility Data

Solubility (In Vitro)

DMSO: 125 mg/mL (220.59 mM) H2O: 1 mg/mL (1.76 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	1.7647 mL	8.8236 mL	17.6473 mL
	5 mM	0.3529 mL	1.7647 mL	3.5295 mL
	10 mM	0.1765 mL	0.8824 mL	1.7647 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.