| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Vitamin A metabolite
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|---|---|
| ln Vivo |
Human plasma was analyzed by high performance liquid chromatography for the presence of retinoic acid and 4-oxoretinoic acid isomers. Peaks that coeluted with the reference compounds all-trans-retinoic acid, 13-cis-retinoic acid, and 13-cis-4-oxoretinoic acid were routinely observed in human plasma. These retinoids were unequivocally identified by the following methods: comigration with reference compounds under several high performance liquid chromatographic conditions; comparison of ultraviolet spectra with those of reference compounds; derivatization with diazomethane and coelution of the methyl esters with reference compounds in a high performance liquid chromatographic system as well as in a gas chromatography system with a mass selective detector. In vitro formation of 13-cis-retinoic acid and 13-cis-4-oxoretinoic acid as artifacts during the analytical procedure was excluded by control experiments. The mean plasma concentrations of the vitamin A metabolites in ten male volunteers were: all-trans-retinoic acid: 1.32 +/- 0.46 ng/ml; 13-cis-retinoic acid: 1.63 +/- 0.85 ng/ml; and 13-cis-4-oxoretinoic acid: 3.68 +/- 0.99 ng/ml. After oral dosing with vitamin A (833 IU/kg body weight) in five male volunteers, mean plasma all-trans-retinoic acid increased to 3.92 +/- 1.40 ng/ml and 13-cis-retinoic acid increased to 9.75 +/- 2.18 ng/ml. Maximal plasma 13-cis-4-oxoretinoic acid concentrations (average 7.60 +/- 1.45 ng/ml) were observed 6 h after dosing which was the last time point in this study. Concentrations of all-trans-4-oxoretinoic acid were low or not detectable. Our findings suggest that, in addition to all-trans-retinoic acid, 13-cis-retinoic acid and 13-cis-4-oxoretinoic acid are present in normal human plasma as metabolites of vitamin A [1].
|
| Animal Protocol |
Plasma samples: Blood was drawn from healthy male volunteers and plasma was prepared by centrifugating the blood in heparinized tubes for 10 min at 1500 g and 4OC. The samples were stored in polypropylene tubes at - 80°C until analysis. Dosing of vitamin A: Volunteers received retinyl palmitate (833 IU/kg body weight) as oily drops [1].
|
| References | |
| Additional Infomation |
This article introduces high-performance liquid chromatography (HPLC), a powerful tool for studying vitamin A metabolism. This method has a limit of quantitation as low as 0.5 ng/ml and can be used to determine the physiological concentrations of 13-cis-4-oxoretinoic acid (13-cis-4-oxoRA), 13-cis-retinoic acid (13-cis-RA), and all-trans-retinoic acid (all-trans-RA) in human plasma. Sample pretreatment using the Varian AASP system is extremely simple, and combined with rapid HPLC elution, it allows for the analysis of large quantities of samples in a short time. Retinol separation was performed using a reversed-phase system with ammonium acetate buffer as the mobile phase. Many researchers commonly use mixtures of aqueous ammonium acetate with acetonitrile or methanol to analyze retinoic acid compounds. We used a 3 μm octadecyl silica column and employed column heating and steep gradient elution to obtain narrow and clear chromatographic peaks. For routine analysis, we used a second UV detector (Shimadzu SPD 6-AV, flow cell: 10 x 1 mm) with a working wavelength of 356 nm, which was connected in series with the first detector with a working wavelength of 340 nm. The output signals of both detectors were processed independently by a C-R4A dual-channel integrator. This made the characteristic UV absorbance ratio of the retinol peak the second basis for peak identification, supplemented by the peak retention time. This detection method has been successfully applied to the study of vitamin A metabolism in human and monkey plasma. [1]
|
| Molecular Formula |
C20H26O3
|
|---|---|
| Molecular Weight |
314.418646335602
|
| CAS # |
71748-58-8
|
| PubChem CID |
6437087
|
| Appearance |
Typically exists as solids at room temperature
|
| Density |
1.07g/cm3
|
| Boiling Point |
509.9ºC at 760 mmHg
|
| Melting Point |
150-157ºC
|
| Flash Point |
276.3ºC
|
| Index of Refraction |
1.565
|
| LogP |
4.782
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
3
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
23
|
| Complexity |
638
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
O=C1C(C)=C(/C=C/C(=C/C=C/C(=C\C(=O)O)/C)/C)C(C)(C)CC1
|
| Synonyms |
4-Keto 13-cis-retinoic acid; 4-Oxoisotretinoin; 71748-58-8; 13-cis-4-Oxoretinoic acid; Oxoretinoic acid; 4-Oxo-13-cis retinoic acid; Ro 22-6595; Retinoic acid, 4-oxo-, 13-cis-; DTXSID201312157; 4-Oxoiisotretinoin; Ro 22-6595
|
| HS Tariff Code |
2934.99.9001
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| 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)
|
| Solubility (In Vitro) |
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
|
|---|---|
| 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 | 3.1805 mL | 15.9023 mL | 31.8046 mL | |
| 5 mM | 0.6361 mL | 3.1805 mL | 6.3609 mL | |
| 10 mM | 0.3180 mL | 1.5902 mL | 3.1805 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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