| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
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| Targets |
Natural isohexenylnaphthazarin pigment
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| ln Vitro |
The phytochemical investigation of the n-hexane extract from callus and cell suspension culture of Arnebia euchroma (Royle) Jonst. resulted in the isolation of nine isohexenylnaphthazarins: deoxyalkannin (1), alkannin (2), acetylalkannin (3), isobutyrylalkannin (4), β-hydroxyisovalerylalkannin (5), 2''-(S)-α-methylbutyrylalkannin (6), propionylalkannin (7), teracrylalkannin (8) and acetylshikonin (9). Their structures were determined by MS and NMR spectroscopy. Pigments 2–8 are isolated for the first time from Arnebia in vitro cultures, 4 and 7 are reported in the present work as novel metabolites within the Arnebia genus, while 9 is a known constituent of both natural roots and in vitro cultures of A. euchroma. Moreover, methyl jasmonate and 1-monoglyceryl olate, palmitate and stearate are reported for the first time within the Boraginaceae family. The antimicrobial and cytotoxic activities of all isolated pigment compounds were tested, revealing a very interesting profile [1].
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| Enzyme Assay |
Antimicrobial Assays [1]
Antimicrobial activity was evaluated using the disc diffusion method by measuring the zone of inhibition. Standard antibiotics netilmicin and 5-flucytocine were used in order to control the sensitivity of the tested bacteria and fungi, respectively. The tested compounds were dissolved in MeOH. For each experiment control disc with pure solvent was used as blind control. All paper discs had a diameter of 6 mm and were deposited on the surface of the seeded trypticase soy agar Petri dishes. The plates were inoculated with the organisms of interest to give a final cell concentration of 107 cells/mL and incubated for 48 h at 37 °C. The fungi were grown on Sabouraud’s agar at 25 °C for 48 h. The experiments were repeated three times and results (diameters in mm) were expressed as mean values. |
| Cell Assay |
Cell Culture [1]
Human leukemia cells CCRF-CEM and breast cancer cells MDA-MB-231 were cultured in RPMI 1640 medium, 2 mM L-glutamine, 10% heat-inactivated fetal bovine serum and 1% Pen/Strep. Human glioblastoma cells U251 and colon cancer cells HCT 116 were grown in Dulbecco’s modified Eagle medium, 2 mM L-glutamine, 10% FBS and 1% Pen/Strep. All cells were kept in a 5% CO2 atmosphere at 37 °C. At 90% confluence cells were passaged. XTT Viability Assay [1] Cell proliferation kit II (XTT) was obtained from Roche Diagnostics. Aliquots (100 µL) of 5 × 104 cells/mL of MDA-MB-231, U251 and HCT 116 cells were seeded in 96-well plates (flat bottom) and grown for 18 h in a humidified 37 °C, 5% CO2 atmosphere before substances were added. Control cells were treated with 0.5% DMSO which had no effect on their growth and viability. In case of CCRF-CEM cells, aliquots (100 µL) of 1 × 105 cells/mL were seeded in 96-well plates (flat bottom) and substances were added immediately. All cells were incubated with the substance of interest for 72 h before XTT solution was added. Vinblastine served as a positive control. XTT is a yellow tetrazolium salt (sodium 3'-[1-(phenylaminocarbonyl)-3,4-tetrazolium]-bis(4-methoxy-6-nitro)benzene sulfonic acid hydrate) and cleaved by metabolic active cells into an orange formazan dye. This colour change occurs only in viable cells and can be directly quantified using a scanning multiwell spectrophotometer. Numbers of viable cells were determined with the following formula expressed as percentage of control: (absorbance of treated cells/absorbance of untreated cells) × 100. IC50 values were calculated using SigmaPlot 11.0 and the four parameter logistic curve. |
| References | |
| Additional Infomation |
The n-hexane extract from cell suspension culture of A. euchroma yielded after several chromatographic separations the compounds deoxyalkannin (1), alkannin (2), acetylalkannin (3), isobutyrylalkannin (4), β-hydroxyisovalerylalkannin (5), 2''-(S)-α-methylbutyrylalkannin (6), propionylalkannin (7) and teracrylalkannin (8) along with methyl jasmonate (10), methyl linoleate, linoleic acid, β-sitosterol, a triglyceride mixture of palmitic, stearic and 8-Ζ- or 9-Ζ- or 11-Ζ or 9-Ε-octadecenoic acid and also a mixture of 1-monoglycerides of palmitic, stearic and oleic acid. Likewise from the n-hexane extract of callus culture of A. euchroma the isohexenylnaphthazarins deoxyalkannin (1), isobutyrylalkannin (4), 2''-(S)-α-methylbutyrylalkannin (6) and acetylshikonin (9) were obtained after several separations. In addition linoleic acid and β-sitosterol were identified after GC-MS analysis of some fractions. Among them 4 and 9 are novel for the Arnebia genus while naphthoquinones 2–8 are isolated from A. euchroma in vitro cultures for the first time. Furthermore 10 and the 1-monoglycerides of palmitic, stearic and oleic acid are probably reported for the first time within Boraginaceae family. As regards the tested antimicrobial activity of 1–9, 2, 4, 5 and 9 where the most active against all the assayed microorganisms while acetylshikonin (9) was found much more active than its enantiomer acetylalkannin (3). Compound 6 exhibited the most potent cytotoxic activity against the tested cancer cell lines while it was estimated that the structure of the side chain or its chirality have a minor effect on their cytotoxic activity. [1]
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| Molecular Formula |
C18H18O6
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| Molecular Weight |
330.332
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| CAS # |
34232-27-4
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| Appearance |
Brown to red solid powder
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| SMILES |
CC(=CCC(C1=CC(=O)C2=C(C=CC(=C2C1=O)O)O)OC(=O)C)C
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| Synonyms |
Acetylalkannin; 34232-27-4; (S)-1-(5,8-dihydroxy-1,4-dioxo-1,4-dihydronaphthalen-2-yl)-4-methylpent-3-enyl acetate
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| 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 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)
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| Solubility (In Vitro) |
DMSO : 100 mg/mL (302.73 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 3.91 mg/mL (11.84 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 39.1 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 (7.57 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.0273 mL | 15.1364 mL | 30.2728 mL | |
| 5 mM | 0.6055 mL | 3.0273 mL | 6.0546 mL | |
| 10 mM | 0.3027 mL | 1.5136 mL | 3.0273 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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