| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg | |||
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| Other Sizes |
Purity: ≥98%
| Targets |
β-lactamase enzymes (hydrolyze the β-lactam ring of
The primary target of nitrocefin is β-lactamase enzymes. Nitrocefin functions as an effective chromogenic substrate due to its unique interaction with β-lactamase enzymes. Upon hydrolysis of the β-lactam ring by β-lactamase, the compound undergoes a structural change that results in a visible color shift from yellow to red. This property enables visual quantification of β-lactamase activity. |
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| ln Vitro |
Michaelis-Menten kinetics of
In bacterial detection assays, Comparison with microtiter plate using UV/vis spectrophotometry gave the same LOD of 10⁶ CFU mL⁻¹ for β-lactamase-expressing E. coli, demonstrating that paper-based In a comparative study of three β-lactamase detection methods, In vitro, nitrocefin is used in biochemical assays to detect β-lactamase enzymatic activity. The compound undergoes hydrolysis by β-lactamase, resulting in a visible color change from yellow to red. The rate of color change is proportional to the enzyme activity, allowing for quantitative measurement. Nitrocefin is used in competitive inhibition studies in the development of β-lactamase-resistant antibiotics. It is also used in clinical microbiology for the rapid detection of β-lactamase-producing bacteria. |
| ln Vivo |
In vivo, nitrocefin is not typically used as a therapeutic agent. It is a diagnostic reagent used for detecting β-lactamase activity in bacterial isolates and clinical samples. The compound is not administered to patients but is used in laboratory settings to identify β-lactamase-producing bacteria that may be resistant to β-lactam antibiotics. Its in vivo applications are limited to diagnostic microbiology.
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| Enzyme Assay |
Reaction optimization on paper: Arrays of 8-mm-diameter paper wells fabricated with filter paper were used. Phosphate buffered saline (PBS) was used as the reaction buffer, and pH was varied to determine optimal pH 7.5. Various concentrations of
Comparison of solution versus dried Nitrocefin test protocol (from Shannon & Phillips): Cell suspensions (10⁸ to 10¹⁰ organisms per mL) were prepared from overnight growth on suitable media. In some cases, cells were subjected to ultrasonic disintegration. General protocols for β-lactamase activity assays using nitrocefin involve preparing a nitrocefin solution in phosphate-buffered saline (PBS) at a concentration of 100-500 μM. The assay is performed in 96-well microtiter plates. Bacterial cultures or purified β-lactamase preparations are added to the wells containing nitrocefin solution. The absorbance at 490 nm (or 485 nm) is measured immediately after addition and at regular intervals (e.g., every minute) for 15-30 minutes using a microplate reader. The rate of absorbance increase is calculated and used to determine β-lactamase activity. The color change from yellow to red is visually observable. |
| Cell Assay |
Detection of β-lactamase-expressing bacteria: Laboratory E. coli expressing β-lactamase were cultured, serially diluted, and mixed with 0.5 mM
Detection in environmental samples: Influent and effluent water samples from a wastewater treatment facility were incubated in growth media for 0-12 hours. Aliquots were taken every 2 hours and tested with Testing bacterial isolates: Ten different bacterial species isolated from environmental samples were tested blindly with In the comparative study (Shannon & Phillips), General protocols for nitrocefin-based detection of β-lactamase-producing bacteria involve preparing bacterial suspensions from colonies on agar plates. A nitrocefin disk or solution is added to the bacterial suspension, and the mixture is incubated at 37°C for 5-15 minutes. A positive result (β-lactamase production) is indicated by a color change from yellow to red. The test is rapid, simple, and widely used in clinical microbiology for the detection of extended-spectrum β-lactamases (ESBLs) and carbapenemases. The results are interpreted visually or using a spectrophotometer for quantitative measurement. |
| Animal Protocol |
General protocols for in vivo detection of β-lactamase activity using nitrocefin are not applicable, as nitrocefin is used exclusively as an in vitro diagnostic reagent. In animal models, nitrocefin can be used to detect β-lactamase-producing bacteria in infected tissues or to evaluate the efficacy of β-lactamase inhibitors. For such studies, tissue samples or abscess fluid are collected and mixed with nitrocefin solution. The color change is observed and quantified. However, these applications are research-oriented rather than clinical.
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| ADME/Pharmacokinetics |
Nitrocefin is a chromogenic substrate and is not administered systemically. Its pharmacokinetic properties are not relevant for its intended use as a diagnostic reagent. The compound has a molecular weight of 516.50 g/mol (C21H16N4O8S2). It is insoluble in ethanol and water but soluble in DMSO at ≥20.24 mg/mL. The compound is typically stored at -20°C. Its stability and handling are well characterized for diagnostic applications.
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| Toxicity/Toxicokinetics |
Nitrocefin is a diagnostic reagent and is not intended for therapeutic use. As a chromogenic substrate, it is considered safe for laboratory use when handled according to standard safety protocols. The compound may cause skin and eye irritation upon contact. Inhalation of dust should be avoided. Appropriate personal protective equipment (gloves, safety goggles, lab coat) should be used. Nitrocefin is not a pharmaceutical drug and has no toxicity data relevant to human exposure.
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| References |
[2]. Utilizing Paper-Based Devices for Antimicrobial Resistant Bacteria Detection. Angew Chem Int Ed Engl. 2017 May 5. |
| Additional Infomation |
Nitrocefin are chromogenic cephalosporin substrates used to detect the presence of β-lactamases, which are important mediators of bacterial antibiotic resistance. While other detection methods exist, such as PCR, nitrocephalosporin assays can be performed rapidly with minimal reagents and inexpensive equipment. Nitrocefin are β-lactam antibiotics with diagnostic properties. They are sensitive to the hydrolytic activity of all β-lactamases produced by Gram-positive and Gram-negative bacteria. The reagent undergoes a color change when its amide bond is hydrolyzed by β-lactamases, thus making it suitable for studying β-lactamase activity.
Background: Applications: Mechanism: The reaction involves hydrolysis of the carbon-nitrogen bond in the β-lactam ring of Nitrocefin is a chromogenic cephalosporin substrate used for the detection of β-lactamase enzymes. It is widely used in clinical microbiology and research laboratories for the rapid detection of β-lactamase-producing bacteria. The compound undergoes a distinctive color change from yellow to red upon hydrolysis by β-lactamase. Nitrocefin is used in competitive inhibition studies in the development of β-lactamase-resistant antibiotics. It has no therapeutic indications and is not approved for clinical use. It is strictly a research and diagnostic reagent. |
| Molecular Formula |
C21H16N4O8S2
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|---|---|
| Molecular Weight |
516.50374
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| Exact Mass |
516.04
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| CAS # |
41906-86-9
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| PubChem CID |
6436140
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| Appearance |
Yellow to orange solid powder
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| Density |
1.7±0.1 g/cm3
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| Boiling Point |
872.0±65.0 °C at 760 mmHg
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| Melting Point |
>99℃
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| Flash Point |
481.2±34.3 °C
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| Vapour Pressure |
0.0±0.3 mmHg at 25°C
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| Index of Refraction |
1.749
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| LogP |
1.04
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
35
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| Complexity |
991
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| Defined Atom Stereocenter Count |
2
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| SMILES |
C1C(=C(N2[C@H](S1)[C@@H](C2=O)NC(=O)CC3=CC=CS3)C(=O)O)/C=C/C4=C(C=C(C=C4)[N+](=O)[O-])[N+](=O)[O-]
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| InChi Key |
LHNIIDJCEODSHA-OQRUQETBSA-N
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| InChi Code |
InChI=1S/C21H16N4O8S2/c26-16(9-14-2-1-7-34-14)22-17-19(27)23-18(21(28)29)12(10-35-20(17)23)4-3-11-5-6-13(24(30)31)8-15(11)25(32)33/h1-8,17,20H,9-10H2,(H,22,26)(H,28,29)/b4-3+/t17-,20-/m1/s1
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| Chemical Name |
(6R,7R)-3-[(E)-2-(2,4-dinitrophenyl)ethenyl]-8-oxo-7-[(2-thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
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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) |
DMSO : ~100 mg/mL (~193.61 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 | 1.9361 mL | 9.6805 mL | 19.3611 mL | |
| 5 mM | 0.3872 mL | 1.9361 mL | 3.8722 mL | |
| 10 mM | 0.1936 mL | 0.9681 mL | 1.9361 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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