NBD-F (4-Fluoro-7-nitrobenzofurazan)
(Synonyms: 4-氟-7-硝基苯并-2-氧杂-1,3-二唑,4-Fluoro-7-nitrobenzofurazan) 目录号 : GC30282
NBD-F (4-Fluoro-7-nitrobenzofuraza) (4-Fluoro-7-nitrobenzofuraza) 是一种前荧光试剂，专为氨基酸分析而开发。
Cas No.:29270-56-2
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Kinase experiment: | An accurately weighed quantity (0.0183 g) of NBD-F is transferred into a 1 mL centrifuge tube, dissolved in acetonitrile and made up to volume with the same solvent to produce stock solutions of 0.1 M. The solution is protected from light and stored at -20°C until analyzed. A 100 μL aliquot of mixed amino acids solution or sample supernatant, 175 μL of borate buffer solution, 200 μL of acetonitrile and 25 μL of NBD-F working solution are mixed in a 1.5 mL centrifuge tube. The well-mixed solution is allowed to react at 60°C in the water bath for 7 min, excluding light. NBD-F reacts with amino group and enables amino acids to be detected with UV detection. After cooling to room temperature, 10 μL of the solution is injected into the equilibrated HPLC system[1]. |
References: [1]. Wu X, et al. Determination of amino acid neurotransmitters in rat hippocampi by HPLC-UV using NBD-F as a derivative. Biomed Chromatogr. 2014 Apr;28(4):459-62. | |
NBD-F is a fluorescent derivatization reagent which is originally developed for amino acid analysis.
As the percentage of the organic phase changes, the retention time of NBD-F remains relatively stable, while the retention times of the derivatization products changes. The pH of the mobile phase affects the separation of the NBD-F and the derivatization products[1]. NBD-F is a fluorescent derivatization reagent that is originally developed for amino acid analysis, and recently applied to the analysis of other amino acid derivatives such as N-methyl-D-aspartic acid and glutathione. The use of NBD-F appears to have several advantages in that the derivatization procedure is simple and its derivatives are highly stable[2].
[1]. Wu X, et al. Determination of amino acid neurotransmitters in rat hippocampi by HPLC-UV using NBD-F as a derivative. Biomed Chromatogr. 2014 Apr;28(4):459-62. [2]. Ishikawa T, et al. Development of an LC-MS/MS method for the analysis of free sphingoid bases using 4-fluoro-7-nitrobenzofurazan (NBD-F). Lipids. 2014 Mar;49(3):295-304.
| Cas No. | 29270-56-2 | SDF | |
| 别名 | 4-氟-7-硝基苯并-2-氧杂-1,3-二唑,4-Fluoro-7-nitrobenzofurazan | ||
| Canonical SMILES | O=[N+](C1=CC=C(F)C2=NON=C21)[O-] | ||
| 分子式 | C6H2FN3O3 | 分子量 | 183.1 |
| 溶解度 | DMSO : ≥ 125 mg/mL (682.69 mM) | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 5.4615 mL | 27.3075 mL | 54.615 mL |
| 5 mM | 1.0923 mL | 5.4615 mL | 10.923 mL |
| 10 mM | 0.5461 mL | 2.7307 mL | 5.4615 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Development of an LC-MS/MS method for the analysis of free sphingoid bases using 4-fluoro-7-nitrobenzofurazan (NBD-F)
The molecular species of sphingoid bases were tagged with the fluorescent amino group reagent, 4-fluoro-7-nitrobenzofurazan (NBD-F). The NBD-sphingoid bases were analyzed by a highly selective and sensitive liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) technique capable of reliable detection of several fmol of the derivatives. Lipid extracts from plant samples were derivatized with NBD-F, and all nine species of free sphingoid bases present in plant sphingolipids were separated and quantified for the first time; a complete baseline resolution was achieved for cis-8 and trans-8 isomers of sphingoid bases by reversed phase HPLC on a C18 column. The extraction and derivatization procedures and LC-MS/MS method can facilitate the progress of the studies for seeking the active components of sphingoid bases species in response to biological challenges.
Quantification of glutathione in plasma samples by HPLC using 4-fluoro-7-nitrobenzofurazan as a fluorescent labeling reagent
A rapid and highly sensitive high-performance liquid chromatograpy method with fluorescence detection has been developed for determination of glutathione (GSH) in human plasma. A simple pre-column derivatization procedure with 7-flouro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-F) reagent was employed. The separation of the derivatized glutathione was performed using a mobile phase consisting of phosphate buffer (0.02 mol/L, pH 6.0)-acetonitrile (77:23, v/v) at a flow rate of 1.0 mL/min with the column temperature 2°C. The eluted derivatives were fluorometrically detected at an excitation wavelength 470 nm and an emission wavelength 530 nm. Under the optimum chromatographic conditions, the calibration curve was linear over the range of 0.1 ?mol/L to 10.0 ?mol/L with the correlation coefficient of 0.9988. The precision of the method was satisfactory with the intra- and inter-day coefficient of variation being 6.3%, 6.9%, respectively. This method has been used to determine glutathione concentrations in plasma samples from healthy individuals.
Highly sensitive determination and validation of gabapentin in pharmaceutical preparations by HPLC with 4-fluoro-7-nitrobenzofurazan derivatization and fluorescence detection
A sensitive HPLC method with pre-column fluorescence derivatization using 4-Fluoro-7-Nitrobenzofurazan (NBD-F) has been developed for the determination of gabapentin in pharmaceutical preparations. The method is based on the derivatization of gabapentin with (NBD-F) in borate buffer of pH 9.5 to yield a yellow, fluorescent product. The HPLC separation was achieved on a Inertsil C(18) column (250 mm × 4.6 mm) using a mobile phase of methanol water (80:20, v/v) solvent system at 1.2 mL/min flow rate. Mexiletine was used as the internal standard. The fluorometric detector was operated at 458 nm (excitation) and 521 nm (emission). The assay was linear over the concentration range of 5 50 ng/mL. The method was validated for specificity, linearity, limit of detection, limit of quantification, precision, accuracy, robustness. Moreover, the method was found to be sensitive with a low limit of detection (0.85 ng/mL) and limit of quantitation (2.55 ng/mL). The results of the developed procedure for gabapentin content in capsules were compared with those by the official method (USP 32). Statistical analysis by t- and F-tests, showed no significant difference at 95 confidence level between the two proposed methods.
Identification of the lysine residue responsible for coenzyme A binding in the heterodimeric 2-oxoacid:ferredoxin oxidoreductase from Sulfolobus tokodaii, a thermoacidophilic archaeon, using 4-fluoro-7-nitrobenzofurazan as an affinity label
The heterodimeric 2-oxoacid:ferredoxin oxidoreductase (StOFOR) from Sulfolobus tokodaii, a thermoacidophilic archaeon, was inactivated by low concentrations of 4-fluoro-7-nitrobenzofurazan (NBD-F), with concomitant increase in fluorescence in subunit-b. The inactivation was prevented by CoA, suggesting that NBD-F covalently bound to the Lys which is responsible for CoA binding. The NBD-labeled subunit-b was isolated and digested with endoproteinase Lys-C. The resulting polypeptide mixture was separated by reverse phase HPLC and the fluorescent fraction was isolated. Amino acid sequencing of the fraction revealed that it comprised a mixture of two polypeptides containing Lys125 and Lys173, respectively. Two StOFOR mutants, K125A and K173A, were constructed, expressed and purified. K125A showed a large increase in the K(m) value for CoA and showed poor inactivation by NBD-F, compared with K173A and wild type StOFOR, indicating Lys125 in subunit-b is the critical residue that interacts with CoA.
Determination of plasma free 3-nitrotyrosine and tyrosine by reversed-phase liquid chromatography with 4-fluoro-7-nitrobenzofurazan derivatization
Oxidative stress plays an important role in pathogenesis of many diseases. Measurement of 3-nitrotyrosine (NO(2)Tyr), as a potential biomarker for nitric oxide-mediated damage, has recently been the focus of particular attention. We have developed an HPLC method with NBD-F pre-column derivatization followed by C(18) cartridge cleaning. Using this method we achieved limits of detection of 0.5 and 1.1 nm for NO(2)Tyr and tyrosine (Tyr), respectively, close to that achieved by LS-MS/MS. NO(2)Tyr and tyrosine concentrations were linear over the calibration ranges 0.5-100 nm and 1-320 microm, respectively, with correlation coefficients greater than 0.95. To evaluate the utility of this assay in plasma we analysed samples obtained from smokers and non-smoking subjects. Consistent with the presence of elevated oxidative stress, the plasma NO(2)Tyr concentration and NO(2)Tyr:Tyr ratio of smokers were 17.42 +/- 11.6 nm and 0.263 +/- 0.192 nm/microm with 3.8 and 3.9 times higher (both p < 0.05), respectively, than that of non-smoker controls (4.54 +/- 2.75 nm and 0.067 +/- 0.050 nm/microm, respectively). In conclusion, we have developed a novel HPLC assay for NO(2)Tyr without MS detection that is applicable to clinical studies addressing the pathophysiology and importance of oxidative stress.