| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
The labeled compound 5-Aminolevulinic acid-13C2,15N hydrochloride does not target specific receptors as an internal standard. Unlabeled 5-Aminolevulinic acid (5-ALA) is an intermediate in heme biosynthesis in the body and the universal precursor of tetrapyrroles (heme, chlorophyll, cobalamin (vitamin B12), and bilirubin). 5-ALA is a natural amino acid derivative that is converted by ALA dehydratase (ALAD) and downstream enzymes to porphobilinogen, hydroxymethylbilane, uroporphyrinogen III, coproporphyrinogen III, protoporphyrin IX, and finally heme (via ferrochelatase). 5-ALA does not have a classical receptor target.
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| ln Vitro |
Compounds labeled with stable or radioactive isotopes can accurately track and quantify individual atoms in metabolic pathways. Stable isotopes generally do not change molecular properties, but may slightly affect metabolic dynamics; radioactive isotopes may interfere with cells. Labeling can distinguish endogenous and exogenous metabolites, reduce false positives, and is beneficial for quantification and reconstruction of metabolic pathways [2]. In cell culture or enzyme reactions, the use of isotope labels can accurately control concentration and exposure time, making it easier to study metabolic reactions and enzyme activities. Through stable isotope-resolved metabolomics (SIRM), cellular metabolic networks can be studied, key metabolic nodes and regulatory mechanisms can be identified, and targets can be provided for compound development. Isotope-labeled compounds can be used in competition binding experiments to evaluate the affinity and binding kinetics of compounds with receptors, which helps optimize the design. Stable isotope labels are used as internal standards in mass spectrometry analysis to improve analytical accuracy and reproducibility and reduce matrix effect interference [3].
In vitro, unlabeled 5-ALA (0.1-10 mM) is used in photodynamic therapy (PDT) research. Exogenous 5-ALA bypasses the rate-limiting step of heme biosynthesis (the feedback inhibition of ALAS1/2 by heme), leading to accumulation of fluorescent porphyrins, primarily protoporphyrin IX (PpIX), in cells. PpIX is a photosensitizer that, upon excitation with blue light (400-410 nm), generates singlet oxygen and ROS, inducing apoptosis and necrosis. This forms the basis for 5-ALA PDT for actinic keratosis, basal cell carcinoma, and bladder cancer diagnosis (blue light cystoscopy). In cancer cells, 5-ALA (0.1-5 mM, 4-24 hours) increases PpIX fluorescence, enabling cell visualization and photosensitization. In mitochondria, 5-ALA enhances heme synthesis and mitochondrial function at low concentrations (1-100 microM). No dedicated in vitro studies exist for 5-Aminolevulinic acid-13C2,15N hydrochloride. |
| ln Vivo |
Isotope labels can non-invasively track the distribution, transformation, and clearance of compounds and their metabolites in the body through techniques such as mass spectrometry (MS) and nuclear magnetic resonance (NMR), which is beneficial for the study of drug metabolism dynamics (ADME). Isotope labeling can reveal specific steps in the metabolic pathway. Direct use of compounds with stable isotope labels at specific positions in human or animal models can also help verify drug mechanisms and evaluate unexpected side effects, improving the accuracy and efficiency of clinical research [3].
In vivo, unlabeled 5-ALA is used clinically: (1) Photodynamic therapy (PDT) for actinic keratosis (20% 5-ALA solution applied topically, 3-4 hours illumination with blue light, 417 nm). (2) Fluorescence-guided resection of malignant gliomas (5-ALA 20 mg/kg oral, 3 hours before surgery). (3) Photodynamic diagnosis of bladder cancer (intravesical 5-ALA). 5-ALA also has potential applications as a dietary supplement (iron absorption enhancer). No dedicated in vivo studies exist for 5-Aminolevulinic acid-13C2,15N hydrochloride; it is used as internal standard for 5-ALA quantitation. |
| Enzyme Assay |
No specific receptor binding protocols exist for 5-Aminolevulinic acid-13C2,15N hydrochloride. For heme biosynthesis enzyme assays: For ALA dehydratase (ALAD): Incubate red blood cell lysate or liver homogenate in 0.1 M Tris-HCl, pH 8.2, containing 5-10 mM ALA (unlabeled) or 5-Aminolevulinic acid-13C2,15N hydrochloride (as internal standard) for 60 minutes at 37degC. Terminate with 10% TCA, centrifuge. Add Ehrlich's reagent (p-dimethylaminobenzaldehyde in glacial acetic acid) to the supernatant, develop color for 15 minutes, measure absorbance at 555 nm. The porphobilinogen formed reacts with Ehrlich's reagent. For analytical quantification of 5-ALA in biological samples using isotope-labeled IS: Prepare calibration standards of unlabeled 5-ALA hydrochloride in blank plasma/urine. Add 5-Aminolevulinic acid-13C2,15N hydrochloride (10-100 ng/mL, final) as internal standard. Derivatize with dansyl chloride or other reagent for LC-MS analysis, or analyze directly by HILIC-MS/MS.
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| Cell Assay |
No dedicated cell-based assay protocols exist for 5-Aminolevulinic acid-13C2,15N hydrochloride. For 5-ALA PDT studies (unlabeled): Culture cancer cells (e.g., HeLa, SCC, U87 glioma) in DMEM with 10% FBS. Treat with 5-ALA hydrochloride (0.1-5 mM) for 4-24 hours in the dark. Wash cells with PBS, add fresh medium, and illuminate with blue light (400-410 nm, 10-100 J/cm2, LED array). Incubate for 24 hours post-illumination. Assess cell viability by MTT assay, apoptosis by Annexin V/PI staining, and ROS production by DCFH-DA fluorescence. For PpIX fluorescence: Wash treated cells with PBS, detach, and analyze by flow cytometry (excitation 405 nm, emission 630-640 nm) or image with fluorescence microscope (excitation 400-410 nm, emission >600 nm). For 5-Aminolevulinic acid-13C2,15N hydrochloride as IS: Spike into cell lysates or culture medium before LC-MS analysis of 5-ALA.
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| Animal Protocol |
No dedicated animal protocols exist for 5-Aminolevulinic acid-13C2,15N hydrochloride. For 5-ALA PK studies (unlabeled) where isotope-labeled IS is used for LC-MS quantitation: Administer 5-ALA hydrochloride (10-100 mg/kg, oral, IV, or IP) to rodents (rats or mice). Collect blood samples at 0 (pre-dose), 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, 24 hours. Centrifuge to obtain plasma. Add 5-Aminolevulinic acid-13C2,15N hydrochloride (10-100 ng/mL) to plasma samples, calibrators, and QCs as internal standard. Derivatize (dansyl chloride or other suitable reagent) or analyze directly by HILIC-MS/MS. Extract by protein precipitation with acetonitrile/methanol (3-5 volumes). Calculate PK parameters (Cmax, Tmax, AUC, t½, CL, Vd). For tissue distribution studies (brain, liver, kidney, tumor for PDT models): Homogenize tissues in methanol/water, add IS, extract, analyze similarly. For fluorescence imaging studies, unlabeled 5-ALA (10-100 mg/kg oral or IV) induces PpIX accumulation in tumors (peak at 3-6 hours). Excise tumors, section, and image for PpIX fluorescence (excitation 405 nm, emission >600 nm).
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| ADME/Pharmacokinetics |
No specific PK data for 5-Aminolevulinic acid-13C2,15N hydrochloride exist. PK properties of unlabeled 5-ALA hydrochloride: Absorption: After oral administration, 5-ALA is absorbed from the small intestine via peptide transporter PEPT1 (Tmax 0.5-1 hour in humans and rodents). Oral bioavailability is variable (20-60%) depending on dose and formulation. For IV administration, Tmax is immediate. Distribution: 5-ALA has low plasma protein binding (<20%). Volume of distribution is moderate (~0.5-1 L/kg). 5-ALA crosses the blood-brain barrier to a limited extent. Metabolism: 5-ALA is not directly metabolized; it is converted to porphobilinogen (by ALA dehydratase, ALAD) and ultimately to heme via the porphyrin pathway. Some 5-ALA is metabolized to 4,5-dioxovaleric acid (DOVA) via non-enzymatic transamination. Excretion: 5-ALA and its metabolites are excreted primarily in urine (60-80% of dose within 24 hours). Renal clearance: 200-400 mL/min in humans (involves glomerular filtration and tubular secretion). Half-life: 0.5-1 hour in humans, 0.5-2 hours in rodents.
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| Toxicity/Toxicokinetics |
No dedicated toxicity data for 5-Aminolevulinic acid-13C2,15N hydrochloride exist. Unlabeled 5-ALA hydrochloride is approved by FDA and EMA for medical use in PDT (actinic keratosis, basal cell carcinoma), fluorescence-guided resection of malignant gliomas (ALA-Gliolan, 20 mg/kg oral), and photodynamic diagnosis of bladder cancer. Acute oral LD50 in rats: >5000 mg/kg. At therapeutic doses (oral 20-100 mg/kg), adverse effects include transient hypotension, nausea, vomiting, elevated liver enzymes, and photosensitivity (for 24-48 hours after administration). At high doses (>200 mg/kg), neurotoxicity (due to porphyrin accumulation in nervous system) may occur, manifesting as neuropathy, seizures, and hyponatremia (due to SIADH). Contraindications: acute porphyria, pregnancy, lactation. The 13C2,15N-labeled form has identical toxicity profile at research-grade quantities. 5-Aminolevulinic acid-13C2,15N hydrochloride is for research use only, not for human use.
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| References | |
| Additional Infomation |
5-Aminolevulinic acid-13C2,15N hydrochloride is a research-use only stable isotope-labeled compound, not approved for diagnostic or therapeutic use in its labeled form (although unlabeled 5-ALA is FDA-approved). It has not been evaluated in clinical trials as an isotopically labeled tracer. Its primary application is as an internal standard for quantitative analysis by NMR, GC-MS, or LC-MS in: (1) Pharmacokinetic and metabolism studies of 5-ALA; (2) Metabolomics studies of heme biosynthesis, porphyrin metabolism, and tetrapyrrole pathways; (3) Quantitation of 5-ALA in plasma, urine, tissues, and food samples; (4) Method development for bioanalytical assays. Unlabeled 5-ALA is an intermediate in heme biosynthesis and a photosensitizer precursor used in photodynamic therapy for cancer and in fluorescence-guided surgery.
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| Molecular Formula |
C313C2H10CL15NO3
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|---|---|
| Molecular Weight |
170.57
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| Exact Mass |
170.038
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| CAS # |
113639-01-3
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| PubChem CID |
168006541
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| Appearance |
Typically exists as solid at room temperature
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
10
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| Complexity |
121
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C(C[13C](=O)[13CH2][15NH2])C(=O)O.Cl
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| InChi Key |
ZLHFONARZHCSET-MBIZCHBVSA-N
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| InChi Code |
InChI=1S/C5H9NO3.ClH/c6-3-4(7)1-2-5(8)9;/h1-3,6H2,(H,8,9);1H/i3+1,4+1,6+1;
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| Chemical Name |
5-(15N)azanyl-4-oxo(4,5-13C2)pentanoic acid;hydrochloride
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| Synonyms |
5-ALA-13C2,15N hydrochloride; δ-Aminolevulinic acid-13C2,15N hydrochloride; 5-Amino-4-oxopentanoic acid-13C2,15N hydrochloride
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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 |
| 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) |
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
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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 | 5.8627 mL | 29.3135 mL | 58.6270 mL | |
| 5 mM | 1.1725 mL | 5.8627 mL | 11.7254 mL | |
| 10 mM | 0.5863 mL | 2.9313 mL | 5.8627 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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