Resonance Rayleigh Scattering Intensity Research Articles

Determination of glycyrrhizic acid as sweeteners by dual-wavelength overlapping resonance Rayleigh scattering

This study used cationic dyes methyl violet (MV) and neutral red (NR) as probes, based on dual-wavelength overlapping resonance Rayleigh scattering (RRS), and designed two highly sensitive methods for the determination of the natural sweetener glycyrrhizic acid (GZA). In the Clark-Lubs buffer solution, when only MV or NR was present in the solution, they formed supramolecular polymers through hydrogen bonding. When adding GZA at different concentrations, the formation of ionic association complexes led to a subsequent decrease in RRS intensity at two wavelengths in the MV − GZA system and NR − GZA system, and the amount of the decline was proportional to the GZA concentration. The linear ranges were determined as 0.1 − 0.6 mg/L and 0.1 − 0.7 mg/L, with correlation coefficients of 0.9972 and 0.9996, and limits of detection were 0.052 mg/L and 0.016 mg/L, respectively. For this study, reaction conditions were optimized, potential interfering substances were investigated, and the mechanisms behind RRS generation and intensity decrease were discussed. The proposed methods were sensitive, fast, time-saving and efficient, and have been successfully applied to licorice and dried ginger decoction analysis, yielding satisfactory results.

Study on the interaction between erythrosine B and the cardiac drug amiodarone using fluorescence, scattering, and absorbance spectra and their analytical application.

In an acidic buffered solution, erythrosine B can react with amiodarone to form an association complex, which not only generates great enhancement in resonance Rayleigh scattering (RRS) spectrum of erythrosine B at 346.5 nm but also results in quenching of fluorescence spectra of erythrosine B at λemission= 550.4 nm/λexcitation= 528.5nm. In addition, the formed erythrosine B-amiodarone complex produces a new absorbance peak at 555 nm. The spectral characteristics of the RRS, absorbance, and fluorescence spectra, as well as the optimum analytical conditions, were studied and investigated. As a result, new spectroscopic methods were developed to determine amiodarone by utilizing erythrosine B as a probe. Moreover, the ICH guidelines were used to validate the developed RRS, photometric, and fluorimetric methods. The enhancements in the absorbance and the RRS intensity and the decrease in the fluorescence intensity of the used probe were proportional to the concentration of amiodarone in ranges of 2.5-20.0, 0.2-2.5, and 0.25-1.75 μg/mL, respectively. Furthermore, limit of detection values were 0.52 ng/mL for the spectrophotometric method, 0.051 μg/mL for the RRS method, and 0.075 μg/mL for the fluorimetric method. Moreover, with good recoveries, the developed spectroscopic procedures were applied to analyze amiodarone in its commercial tablets.

Rapid and selective RRS determination of ferrocyanide with a nanogold surface molecularly imprinted polymethacrylic acid probe.

In this work, a nanogold surface molecularly imprinted polymer spectral probe (AuNP@MIP) for selectively identifying ferrocyanide was prepared under microwave irradiation using nanogold as the core, ferrocyanide as the template ion, methacrylic acid as the monomer, and ethylene glycol dimethacrylate as the cross-linking agent. AuNP@MIP was found to produce a resonance Rayleigh scattering (RRS) peak at 370 nm. When potassium ferrocyanide (K4Fe(CN)6) was present, a AuNP@MIP-Fe(CN)6 complex was formed, producing RRS-energy transfer (RRS-ET). With an increase in ferrocyanide concentration within a certain range, the RRS intensity at 370 nm decreased linearly, and the detection range was 0.02-0.40 μmol L-1, with a detection limit as low as 0.006 μmol L-1 ferrocyanide. This new method has the advantages of simplicity, rapidity, and selectivity when applied for the determination of K4Fe(CN)6 in table salt.

Antibody-labeled gold nanoparticle based resonance Rayleigh scattering detection of S100B.

Traumatic brain injury (TBI) is a sudden brain injury due to an external force that causes a large number of deaths and permanent disabilities every year. S100B has been recognized as a potential objective quantitative biomarker for screening the prognosis of TBI and severe head injury. In this article, an anti-S100B monoclonal antibody was immobilized on cysteamine (Cy) functionalized gold nanoparticles (AuNPs) by EDC-NHS chemistry, which enabled S100B resonance Rayleigh scattering (RRS) detection based on antibody-labeled gold nanoparticles. The prepared conjugates were characterized by ultraviolet-visible spectroscopy (UV-vis), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), dynamic light scattering (DLS) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Based on the specific binding of the antibody and antigen, the RRS intensities at 381 nm and 541 nm wavelengths were significantly enhanced, and thus a dual wavelength overlapping resonance Rayleigh scattering (DWO-RRS) method was established. The scattering intensity of the two overlapping peaks was proportional to the concentration of S100B in the range of 0.05-4.5 ng mL-1 with a detection limit of 0.002 ng mL-1. The proposed DWO-RRS method is time-saving, simple, sensitive, and can be used to determine the concentration of S100B in human serum with satisfactory results, which has a promising application in the early diagnosis of TBI.

Integrated resonance Rayleigh scattering approach utilizing Box-Behnken experimental design for the facile quantification of prucalopride in pharmaceutical tablets and human urine with sustainability assessment.

Prucalopride (PCP) is one of the recent drugs used for the regulation of gastrointestinal tract motility and the treatment of constipation. A new, highly sensitive and fast resonance Rayleigh scattering (RRS) approach was suggested for PCP determination. The approach was based on its reaction of PCP with eosin Y in buffered medium (pH 3.5) to form an ion pair association complex which had a significant enhancement in RRS compared to that of eosin Y or PCP alone. The enhancement of RRS intensity had straight correlation to PCP concentration ranging from 150 to 2000 ng mL-1 with 38 ng mL-1 as LOD and 125 ng mL-1 as LOQ. The measurements were done at a wavelength of 365 nm that provided the maximum sensitivity. All the experimental parameters were studied carefully and optimized via Box-Behnken experimental design. The International Council for Harmonization (ICH) guidelines were employed to validate the suggested method and the obtained results proved the appropriate method performance. The method was efficiently utilized to determine PCP in pure form, pharmaceutical tablets and spiked urine samples with no interferences from the surrounding matrices. Furthermore, the greenness of the suggested procedure was confirmed using different green metric approaches.

Aptamer SERS and RRS determination of trace lead ions using nitrogen-doped carbon dot to catalyze the new nano-gold reaction

Nitrogen-doped carbon dots (CDN) were prepared by microwave hydrothermal method using ammonium citrate (AC) and ethylenediaminetetraacetic acid (EDTA) as precursor. It was characterized by transmission electron microscopy (TEM) and infrared spectroscopy (IR). The CDN was found to catalyze the reduction of HAuCl4 to produce gold nanoparticles (AuNP), among which fructose was an effective reducing agent. Using malachite green (MG) as a molecular probe, the surface enhanced Raman scattering (SERS) intensity at 1617 cm−1 and the resonance Rayleigh scattering (RRS) intensity at 375 nm increased linearly with increasing CDN concentration, respectively. The catalytic activity of CDN is inhibited because the aptamer (Apt) can be adsorbed on the surface of the catalyst CDN. The aptamer (Apt)-Pb2+ reaction and CDN-Apt adsorbing reaction were competitive reaction. When there is Pb2+ that binds more tightly to Apt, Apt is desorbed, and the catalytic ability of CDN is restored. Accordingly, an Apt-mediated nanocatalytic amplification SERS/RRS platform for quantitative detection of lead ions was constructed. For the SERS method, the linear range was 0.5–120 nmol/L with DL of 0.11 nmol/L. For the RRS method, the Pb2+ concentration was linear in the range of 50–400 nmol/L with the RRS intensity, and the DL was 15.32 nmol/L. The analysis platform uses CDN catalyzed nanoreactions to generate AuNP products with SERS activity as a substrate, thus overcoming the shortcomings of Pb2+ without scattering activity, and realizing the possibility of SERS and RRS detection of metal ions. It was used for the determination of Pb2+ in real samples with relative standard deviations were 0.94–2.71 % and recovery was 99.00–103.70 %, respectively. In addition, the mechanism of CDN nanoenzyme heterogeneous catalysis of nano-gold reactions was discussed.

A stable and sensitive Au metal organic frameworks resonance Rayleigh scattering nanoprobe for detection of SO3 2- in food based on fuchsin addition reaction.

A stable Au metal organic frameworks (AuMOF) nanosol was prepared. It was characterized by electron microscopy and molecular spectral techniques. In pH 6.8 PBS buffer solution, AuMOF nanoprobes exhibit a strong resonance Rayleigh scattering (RRS) peak at 330 nm. After basic fuchsin (BF) adsorbing on the surface of AuMOF, the RRS energy of the nanoprobe donor can be transferred to BF receptor, resulting in a decrease in the RRS intensity at 330 nm. Both sulfite and BF taken place an addition reaction to form a colorless product (SBF) that exhibit weak RRS energy transfer (RRS-ET) between AuMOF and SBF, resulting in the enhancement of the RRS peak. As the concentration of SO3 2-increases, the RRS peak is linearly enhanced. Thus, a new and sensitive RRS-ET method for the detection of SO3 2- (0.160-5.00 μmol/L) was developed accordingly using AuMOF as nanoprobes, with a detection limit of 0.0800 μmol/L. This new RRS method was applied to determination of SO3 2- in food and SO2 in air samples. The recoveries of food and air samples were 97.1-106% and 92.9-106%, and the relative standard deviation (RSD) was 2.10-4.80% and 2.10-4.50%, respectively.

Applying resonance Rayleigh scattering and spectrofluorimetric techniques for the selective determination of neomycin sulfate using Congo red as a probe: Applications for wastewater analysis.

Two simple, sensitive, and low-cost fluorescence spectroscopy methods for neomycin (NEO) detection were developed. Both methods were based on the interaction between NEO and Congo red (CR) in acidic buffer medium to form an ion-association complex. The quenching effect of the formed ion-association complex on the fluorescence of CR at 421 nm is a basic principle of fluorescence analysis, whilst the resonance Rayleigh scattering (RRS) method was used to enhance the resonance Rayleigh scattering spectrum at 384 nm by adding NEO. Experimental conditions such as pH, temperature, reaction time, CR concentration, and the ionic strength of the two methods were investigated and optimized. In addition, the effect of common coexisting substances on the method was tested and the results showed good selectivity. The composition of ion-association complexes, the reaction mechanism, and reasons for the enhanced intensity of RRS are discussed. Under optimum conditions, the responses of the fluorescence spectrophotometry and RRS methods showed good linearity with NEO concentrations in the range 0.2-3.0μg ml-1 and 0.1-3.0μg ml-1 , respectively. The detection limits of fluorescence spectrophotometry and RRS spectroscopy techniques were 0.02 μg ml-1 and 0.01 μg ml-1 , respectively. Finally, the two methods were applied to the analysis of wastewater and the results were satisfactory.

A new CaMOF resonance Rayleigh scattering probe for trace sulfide based-methylene blue receptor

The as-preared Ca metal organic framework (CaMOF) nanosol exhibits a strong resonance Rayleigh scattering (RRS) peak at 380 nm under the chosen conditions. The S2− reacts with p-aminodimethylaniline (DMPD) to produce methylene blue (MB) in the presence of Fe(III). The produced MB acts as a RRS receptor and CaMOF as RRS donor, the both taken place RRS-energy transfer (RRS-ET) when MB was adsorbed on the CaMOF surface to result in the RRS signal quenching. As the concentration of Na2S increasing, the MB formed increased and the RRS-ET enhanced, leading to a linear decreasing in the RRS intensity at 380 nm. The decreased value ΔI shows a good linear relationship with the concentration of Na2S in the range of 0.050–5.0 μmol/L. Accordingly, a new RRS-ET method was established to determine sulfate in water and H2S in air samples, with high sensitivity, good selectivity and simplicity.

Resonance Rayleigh scattering approach based on association complex formation with erythrosine B for determination of venlafaxine, application to the dosage form and spiked human plasma.

The interaction of venlafaxine hydrochloride (VLX) with erythrosine B was investigated using a resonance Rayleigh scattering (RRS) spectroscopic technique. In acetate buffer (pH3.4), erythrosine B reacted with VLX to form a 1:1 ion-pair complex with concomitant enhancement in RRS intensity that was measured at 330 nm. In addition, the stability constant and the change in free energy of the reaction were estimated. Based on this interaction a new method was developed for a sensitive VLX analysis using erythrosine B as a probe. The results indicated that this method had good selectivity in the presence of coexisting compounds. The scattering intensity (ΔIRRS ) was linearly dependent on VLX concentration over the range 0.04-1.0μg ml-1 with a determination coefficient (r) of 0.9998. The limit of detection and limit of quantitation were 0.01 and 0.03 μg ml-1 , respectively. This method could be suitably used for analysis of VLX in pharmaceutical capsules and human plasma.

Label-Free Resonance Rayleigh Scattering Amplification for Lipopolysaccharide Detection and Logical Circuit by CRISPR/Cas12a-Driven Guanine Nanowire Assisted Non-Cross-Linking Hybridization Chain Reaction.

Although the CRISPR/Cas system has pioneered a new generation of analytical techniques, there remain many challenges in developing a label-free, accurate, and reliable CRISPR/Cas-based assay for reporting the levels of low abundance biomolecules in complex biological samples. Here, we reported a novel CRISPR-derived resonance Rayleigh scattering (RRS) amplification strategy and logical circuit based on a guanine nanowire (G-wire) assisted non-cross-linking hybridization chain reaction (GWancHCR) for label-free detection of lipopolysaccharide (LPS). In the presence of a target, the protospacer-adjacent motif-inserted aptamer is rationally designed to specifically combine with LPS rather than Cas12a, suppressing the trans-cleavage activity of CRISPR/Cas12a and retaining the reporter probes to trigger non-cross-linking aggregation. Owing to the automatic hybridization chain reaction (HCR), in the presence of Mg2+, the released G-quadruplex sequence aggregated to assemble the G-wire superstructure through non-cross-linking. As a result, a dramatically amplified RRS intensity is observed, allowing for reporting LPS levels in a low detection limit of 0.17 pg/mL and a wide linear range among 1.0-100.0 ng/mL. Moreover, this reaction event is capable of programming to perform classical Boolean logic tree analysis, including basic logic computing and complex integrated logic circuits. This study comprehensively analyzed with respect to information flow, matter (molecular events), and energy (RRS), revealing the potential promise in designing of molecular-level "Internet of Things", intelligent computing, and sensing systems.

Polyvinyl alcohol/polyacrylamide hydrogel-based sensor for lead (II) ion sensing by resonance Rayleigh scattering

In this research, a hydrogel based sensor was designed for detection and measurement of lead (II) ions based on resonance Rayleigh scattering (RRS) using polyvinyl alcohol (PVA), and acrylamide (AAm) loaded by Eriochrome Black T (EBT), prepared via in-situ radical polymerization method. The results indicated that the designed sensor is highly selective and sensitive in detecting Pb2+quantitatively. The RRS intensity of the sensor is significantly increased at pH = 11. Under the optimum conditions, the linear detection range was validated from 100 μM to 500 μM. The lead ions recovery from aqueous solution was found in the range of 99% to 104%. The hydrogel/EBT sensor has a potential for sensitive and selective detection of various metal ions or metal based target molecules by changing the selective ligand of the probe in the hydrogel.

A novel resonance Rayleigh scattering assay for trace formaldehyde detection based on Ce-MOF probe and acetylacetone reaction

Formaldehyde (HCHO) is a widespread toxic pollutant, and the content of formaldehyde in food directly affects human health. Therefore, it is particularly important to establish and develop a rapid and accurate method for the determination of trace formaldehyde to ensure food safety. Metal-organic framework (MOF) has the characteristics of both organic and inorganic materials, which can be applied in various fields. What's more, Cerium (Ce) is the most abundant rare earth element. So far, it has no report about using Ce-MOF as a molecular probe of resonance Rayleigh scattering (RRS). The purpose of this research is to establish a new method for formaldehyde by combining the highly selective acetylacetone reaction with the sensitive RRS technology. When formaldehyde is added, acetylacetone reacts with it to form a complex (DDL), which acts as an accepter. There is a RRS-energy transfer (ET) between the DDL and Ce-MOF (act as the donor) because of spectral overlap. As a result, the RRS intensity of the Ce-MOF system decreased with formaldehyde concentration increase. So, Ce-MOF is used as a molecular probe of RRS to detect formaldehyde. In this study, the detection limit of formaldehyde was 0.063 µmol/L, and the determination range was 0.067–66.67 µmol/L. Compared with other methods, this method is simple in pretreatment and operation, and has a lower detection limit. Thus, a convenient detection of trace formaldehyde is realized.

A new mechanism for resonance Rayleigh scattering detection of minoxidil based on catalytic oxidation of silver nanoparticles

This work presents a new method for minoxidil detection based on silver nanoparticle (AgNP) oxidation. Minoxidil, which is a pyrimidine N-oxide, can be reduced to its corresponding pyrimidine via a redox reaction. In this system, acetate buffer serves as a proton source. AgNPs act as electron donors that contribute electrons to the reaction, producing Ag+. Consequently, the sizes and numbers of AgNPs in the system decrease, which results in a decline in their resonance Rayleigh scattering (RRS). By monitoring the RRS intensity at 409 nm, a change in intensity was linearly related to the minoxidil concentration over a concentration range of 0.5 – 5.0 mM. The detection limit was 0.35 mM. This approach is simple and rapid. It is done by directly mixing the drug and AgNPs in an acidic buffer. The reaction was completed within 2 min. This proposed method was successfully utilized for quantification of minoxidil in topical hair-growth formulations.

PVP-coated silver nanocubes as RRS probe for sensitive determination of Haloperidol in real samples

Polyol synthesis of silver nanocubes (Ag NCs) under dark conditions yielded nanoparticles with high uniformity and purity, as well as edge lengths of 42 nm with good stability and scattering cross-section. These nanoparticles were characterized by SEM, TEM, and Uv–vis spectroscopy. The presence of polyvinylpyrrolidone (PVP) as a capping agent on the surface of Ag NCs, as well as its satisfactory interaction level with Haloperidol (Hp) as an antipsychotic drug, has led to the use of these nanoparticles as Resonance RayleighScattering (RRS) probe to measure Hp. Indeed, Hp resulted in reducing the RRS signal of Ag NCs, and this change in RRS intensity was linear in the range of 10.0 to 800.0 µg L-1 of Hp. The limits of detection (LOD) and quantification (LOQ) were found to be 1.5 and 5.0 µg L-1, respectively. The influence of interfering species was studied, and it was found that the suggested method has good selectivity and can be used to monitor Hp in actual samples. As a result, this RRS probe operated well in determining Hp in pharmaceutical and human plasma samples with satisfactory recovery.

A cholesterol benzoate RRS probe for the determination of trace ammonium ions

The measurement of NH4+ has attracted considerable attention with the increase of NH4+ emissions in sewage caused by human activities. So far, a variety of photometric and fluorescence methods for the detection of NH4+ have been researched and summarized, but there is no report about the use of liquid crystals (LCs) cholesteryl benzoate (CB) as a resonance Rayleigh scattering (RRS) probe to determine ammonium ions. In the NaAc-HAc buffer solution with pH = 4.80, the yellow compounds 3,5 diacetyl-1,4 dihydrolutidine (DDL) generated by the reaction of NH4+ with acetylacetone (AT) and formaldehyde (HCHO) act as the energy receiver and CB as the donor. Because the RRS spectrum of CB overlaps with the DDL absorption spectrum, resonance Rayleigh scattering energy transfer (RRS-ET) occurs. When the NH4+ concentration increased, the generated DDL increased, and the RRS-ET also increased, so the RRS intensity of the system at 395 nm decreased. For this reason, a fast and sensitive CB RRS-ET method was established to apply to the detection of NH4+ in water. The detection range was 1.00 × 10−3 − 4.66 μg/mL, and the detection limit was 6.62 × 10−3 μg/mL. Using this method to analyze and detect NH4+ in environmental water samples, the precision and recovery rate were between 1.30–9.30% and 95.5–109.9%, respectively. Therefore, this method has the advantages of sensitivity and simplicity.

A resonance Rayleigh scattering method for sensitive detection of chitosan based on supramolecular complex and mechanism study

A convenient and sensitive resonance Rayleigh scattering (RRS) method for the detection of chitosan (CTS) has been developed via forming Cu-Zn supramolecular complex by complexation reaction, hydrophobic force and electrostatic attraction. The microstructure of the complex was characterized by FT-IR, zeta potential, scanning electron microscope (SEM), UV–vis and RRS. Furthermore, the interaction mechanism among Cu(II), Zn(II), CTS and sodium dodecyl benzene sulfonate (SDBS) was studied. The results revealed that CTS and Cu(II) or Zn(II) formed a supramolecular complex with RRS enhancement in weak acid condition. In the presence of SDBS, the RRS intensity of CTS-Cu(II)-SDBS or CTS-Zn(II)-SDBS was significantly higher than that of the binary system without SDBS at the same CTS concentration. The RRS intensity of CTS-Cu(II)-Zn(II)-SDBS was higher than that of CTS-Cu(II)-SDBS and CTS-Zn(II)-SDBS. The RRS intensity increased linearly with the increase of CTS concentration made it possible to determine CTS quantitatively. In the range extending from 0.10 to 5.00 μg/mL, the equation of linear regression was ΔI=1848.8c-138.3 with a correlation coefficient 0.9996, and the detection limit was estimated to be 37.96 ng/mL. The study was successfully applied for the determination of CTS in health food samples, suggesting its great potential toward CTS analysis.

Sensitive Aptamer SERS and RRS Assays for Trace Oxytetracycline Based on the Catalytic Amplification of CuNCs.

A new method for the determination of oxytetracycline (OTC) has been established by coupling the catalytic amplification reaction of copper nanoclusters (CuNCs) with the aptamer reaction. CuNCs prepared by a wet chemical method have the catalytic activity for the formation of gold nanoparticles (AuNPs) resulting from a HAuCl4-ethanol (En) reaction. The experimental results showed that OTC aptamer (Apt) can be adsorbed on the surface of CuNCs in a non-specific way, thus inhibiting its catalytic activity. When OTC was added to the solution, the OTC-Apt complex was generated by a specific reaction, which made the CuNCs desorb and restore their catalytic activity. With the increase of OTC, the recovery of the catalytic activity of CuNCs is strengthened, the reaction speed is accelerated, and the number of AuNPs is increased. The generated AuNPs exhibited surface enhanced Raman scattering (SERS) signals at 1615 cm−1 in the presence of Vitoria blue 4R (VB4R) molecular probes, and a resonance Rayleigh scattering (RRS) peak at 586 nm. There is a good linear relationship between the intensities of SERS, or RRS, and OTC concentration at the range of 37.5–300 ng/L or 37.5–225 ng/L, respectively. A new SERS and RRS assay for the determination of trace OTC based on the regulation of CuNCs catalysis was established.

Catalytic oxidation of O-phenylenediamine by silver nanoparticles for resonance Rayleigh scattering detection of mercury (II) in water samples

In this study, a facile nanoparticle catalytic sensor for resonance Rayleigh scattering quantification of mercury (II) ion was developed. The developed approach is relied on the selective inhibition of the peroxidase-like activity of polyvinylpyrrolidone-stabilized silver nanoparticles (PVP-Ag-NPs) by mercury (II) ions. The synthesized PVP-Ag-NPs oxidize the aqueous solution of O-Phenylenediamine (colorless) to 2,3-phenazinediamine (bright yellow) and their resonance Rayleigh scattering (RRS) activity was completely suppressed. When mercury (II) was introduced, the RRS activity of PVP-Ag-NPs was turned on combined with a reduction of the intensity of the yellow color. The enhancement in the RRS intensity was related to the concentration of mercury (II) in the linear range of 10–2000 nM. The smaller size (4.5 nm), the large surface area and the uniform size (PDI = 0.379) of the synthesized PVP-Ag-NPs offered a higher chance for interaction between mercury (II) and PVP-Ag-NPs with the advantages of high sensitivity (LOD = 4 nM) and excellent selectivity for mercury (II) detection over several metals and anions.

Micellar sensitized Resonance Rayleigh Scattering and spectrofluorometric methods based on isoindole formation for determination of Eflornithine in cream and biological samples

α-Difluoromethylornithine or Eflornithine is an FDA-approved drug used for the treatment of Sleeping Sickness (as vials dosage form) and also used for diminishing the unwanted excess facial hair in the hirsutism (as creams dosage form). The proposed work is based on the condensation interaction between the amino moiety of Eflornithine and O-phthalaldehyde/2-mercaptoethanol to form a highly fluorescent isoindole derivative. The fluorescence and the Resonance Rayleigh Scattering (RRS) intensities of the reaction product were greatly augmented upon the addition of hexadecyl-trimethyl ammonium bromide by 153% and 250%, respectively. After optimization of the reaction conditions, the formed isoindole derivative was measured fluorometrically at λemission= 429 nm after λexcitation= 337 nm. Moreover, the significant augmentation in the RRS intensity of the formed product was measured at λmax= 422 nm. In regards to accuracy, sensitivity, robustness and precision, the proposed methods were validated according to ICH guidelines. Furthermore, the proposed methods were successfully applied for the assay of Eflornithine in various commercial brands of the pharmaceutical cream samples with good recovery. In addition to the current fluorometric method was confirmed to be effective in the assaying of Eflornithine in spiked plasma and urine specimens with good recovery.