| Size | Price | |
|---|---|---|
| Other Sizes |
| ln Vivo |
A method was developed and validated for the quantification of total 6-Thioguanosine (6-TG) in peripheral blood mononuclear cells (PBMCs) from patients with inflammatory bowel disease (IBD) receiving azathioprine maintenance therapy. [1]
In PBMCs from 8 patients, median total 6-TG concentration was 375 ng/mL (IQR 212 – 1357 ng/mL). The unphosphorylated fraction of 6-TG was below the limit of quantification in 4 out of 8 samples, indicating that 6-TG is present mainly as phosphorylated nucleotides (approximately 10-fold higher than the unphosphorylated form). [1] |
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| Cell Assay |
PBMCs were extracted from patient blood (approximately 12 mL) using density gradient centrifugation. After separation, cells were washed, and remaining erythrocytes were lysed with an ammonium salts solution. Cell count and mean cell volume (MCV) were determined using a cell counter. Aliquots of 1×10⁶ cells were processed for analysis. [1]
For quantification of total (phosphorylated and non-phosphorylated) 6-Thioguanosine, one aliquot of PBMCs was treated with acid phosphatase (0.5 units) to convert phosphorylated metabolites to the unphosphorylated nucleoside form. An untreated aliquot was used to determine the unphosphorylated fraction. After enzymatic digestion, proteins were precipitated with acetonitrile, and the supernatant was dried and reconstituted for UPLC-MS/MS analysis. [1] The calibration curve for 6-TG in PBMCs ranged from 0.0488 to 25 ng per aliquot, with a mean regression coefficient r² of 0.998. The limit of quantification (LOQ) was 0.0488 ng, and the limit of detection (LOD) was 0.0244 ng. Intra- and inter-day precision and accuracy met FDA guidelines. [1] |
| References | |
| Additional Infomation |
2-Amino-9-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)-2-oxacyclopentanoyl]-3H-purine-6-thione is a purine nucleoside.
6-Thioguanosine (6-TG) is a biologically active ribonucleoside metabolite of azathioprine. It is believed to be responsible for the immunosuppressive effect of azathioprine therapy. Overproduction of 6-TG can lead to medullary suppression (excessive effect). [1] This study reports the first UPLC-MS/MS method for quantifying 6-TG in PBMCs, which may serve as a tool for therapeutic drug monitoring at the active site. [1] |
| Molecular Formula |
C10H13N5O5S
|
|---|---|
| Molecular Weight |
315.3
|
| Exact Mass |
299.069
|
| CAS # |
85-31-4
|
| Related CAS # |
85-31-4;345909-25-3 (hydrate);
|
| PubChem CID |
2724387
|
| Appearance |
Off-white to light yellow solid powder
|
| Density |
2.18 g/cm3
|
| Boiling Point |
756.9ºC at 760 mmHg
|
| Melting Point |
230-231°C
|
| Flash Point |
411.6ºC
|
| LogP |
-1.2
|
| Hydrogen Bond Donor Count |
5
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
20
|
| Complexity |
449
|
| Defined Atom Stereocenter Count |
4
|
| SMILES |
C1=NC2=C(N1[C@H]3[C@@H]([C@@H]([C@H](O3)CO)O)O)NC(=NC2=S)N
|
| InChi Key |
OTDJAMXESTUWLO-UUOKFMHZSA-N
|
| InChi Code |
InChI=1S/C10H13N5O4S/c11-10-13-7-4(8(20)14-10)12-2-15(7)9-6(18)5(17)3(1-16)19-9/h2-3,5-6,9,16-18H,1H2,(H3,11,13,14,20)/t3-,5-,6-,9-/m1/s1
|
| Chemical Name |
2-amino-9-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-3H-purine-6-thione
|
| Synonyms |
6-Thioguanosine NSC 29422 NSC29422NSC-29422
|
| 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)
|
| Solubility (In Vitro) |
DMSO : ~20 mg/mL (~66.82 mM)
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| 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 | 3.1716 mL | 15.8579 mL | 31.7158 mL | |
| 5 mM | 0.6343 mL | 3.1716 mL | 6.3432 mL | |
| 10 mM | 0.3172 mL | 1.5858 mL | 3.1716 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.
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