Utility of Gold Nanoparticles for Spectrofluorimetric and Spectrophotometric Determination of Ceftobiprole in Dosage form and Biological Fluids

Subject Areas : Journal of Nanoanalysis

Hesham Salem ¹ , Mahmoud Omar ² , Dina Zein ³ , Deena Mokhater ⁴

1 - faculty of pharmacy, Deraya university, minia, Egypt
2 - Faculty of pharmacy, Minia University
3 - Faculty of pharmacy, Deraya University
4 - Minia University

Received: 2021-08-27 Accepted : 2022-01-26 Published : 2023-02-15

Keywords: Gold nanoparticles, Spectroscopy, Ceftobiprole, dosage forms, biological fluids,

Abstract :

Two simple, fast and novel delicate fluorimetric and spectrophotometric strategies were examined for the measure of ceftobiprole (CEF) utilizing gold nanoparticles (Au NPs). On spectrofluorimetric strategy, gold nanoparticles were utilized as a fluorescence test. The expansion of CEF to Au-NPs arrangement caused significant quenching of the outflow band of Au-NPs, which was likely due to the complexation of the medicate to gold NPs. Beneath the ideal conditions, the extinguished fluorescence (FL) escalated was straight with the examined concentration. The extinguishing instrument of CEF on the outflow band of Au-NPs was clarified by Stern-Volmer law. The moment spectrophotometric strategy was based on conglomeration of synthesized gold nanoparticles. Gold nanoparticles appeared retention at 522 nm. Upon interaction with the CEF, the band at 522 nm vanished with arrangement of a unused ruddy moved band at 673. Distinctive exploratory variables were optimized for higher affectability. The calibration bands were straight with concentration extend of 0.1-12 µg/mL for the examined medicate. The methods were connected effectively to decide the studiedddrug in minor concentrations in immaculate frame, pharmaceutical measurement shapes and organic liquids (human serum and urine samples).

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Full-Text:



J. Nanoanalysis., 10(1): 401-413, Winter 2023

 

 

Utility of Gold Nanoparticles for Spectrofluorimetric and Spectrophotometric Determination of Ceftobiprole in Dosage Form and Biological Fluids

Hesham Salem *,1, Mahmoud A.Omar 2,3, Dina Z. Mazen 1, Deena A. M.Nour El-Deen 2

1       Pharmaceutical Chemistry Department, Faculty of Pharmacy, Deraya University, New Minia, Egypt

2               Analytical Chemistry Department, Faculty of Pharmacy, Minia University, Minia, Egypt

3       Department of Pharmacognosy and Pharmaceutical Chemistry, College of Pharmacy, Taibah University, Medinah, Saudi Arabia

ARTICLE INFO Article History: Received 2021-08-27 Accepted 2022-01-26 Published 2023-02-15   Keywords:  Ceftobiprole, Gold nanoparticles, Spectroscopy, Dosage forms, Biological fluids

ABSTRACT   Two simple, fast and novel delicate fluorimetric and spectrophotometric strategies were examined for the measure of ceftobiprole (CEF) utilizing gold nanoparticles (Au NPs). On a spectrofluorimetric strategy, gold nanoparticles were utilized as a fluorescence test. The expansion of the CEF to Au-NPs arrangement caused significant quenching of the outflow band of Au-NPs, which was likely due to the complexation of the medicate with gold NPs. Under ideal conditions, the extinguished fluorescence (FL) increased straight with the examined concentration. The extinguishing instrument of CEF on the outflow band of Au-NPs was clarified by Stern-Volmer law. The moment spectrophotometric strategy was based on the conglomeration of synthesized gold nanoparticles. Gold nanoparticles appeared to have retention at 522 nm. Upon interaction with the CEF, the band at 522 nm vanished with the arrangement of an unused, ruddy-moved band at 673. Distinctive exploratory variables were optimized for higher affectability. The calibration bands were straight with a concentration range of 0.1-12 µg/mL for the examined medication. The methods were connected effectively to determine the studied drug in minor concentrations in an immaculate frame, pharmaceutical measurement shapes, and organic liquids (human serum and urine samples).

How to cite this article Salem H., Omar M.A., Zein D.Z., El-Deen D.A.N.., Utility of Gold Nanoparticles for Spectrofluorimetric and Spectrophotometric Determination of Ceftobiprole in Dosage Form and Biological Fluids. J. Nanoanalysis., 10(1): 401-413, Winter 2023. DOI: 10.22034/jna.2022.1938902.1271.

 

 

INTRODUCTION

 

Ceftobiprole (CEF) may be a modern 5th-generation cephalosporine utilized to treat hospital-acquired pneumonia (HAP, barring ventilator-associated pneumonia, VAP) and community-acquired pneumonia (CAP) [1]. It may be a beta-lactam anti-microbial operator that has powerful bactericidal action by binding to penicillin-binding protein (PBP), restraining transpeptidation and arrangement of the bacterial cell divider, driving cell lysis, and passing. Ceftobiprole medocaril has broad-spectrum action against gram-positive and gram-negative pathogens that cause complicated diseases [2, 3]. Ceftobiprole is (6R,7R)-7-[[(2Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-hydroxyiminoacetyl] amino]-8-oxo-3-[(E)-[2-oxo-1-[(3R)-pyrrolidin-3-yl]pyrrolidin-3-ylidene]methyl]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (Figure 1) [4]. This sedate was dissected some time recently utilizing HPLC [5] and spectroscopic methods for its examination with Eosin Y of various forms in its pure or pharmaceutical dosage forms [6] and in human plasma, as it were, with no other detailed strategies. Fluorimetric strategies are becoming increasingly vital for quantitative assurance of pharmaceutical drugs [7-9] due to their high affectability, brief run time, cheap instrumentation, and selectivity. The later utilization of luminescent NPs (ZNS, ZnSe, CDS, CdTe, and CdSe) revolutionized expository methods. Medicate assurance due to their interesting properties, which start with their quantum estimate impact, brightness, solid steadiness against photo fading, and resistance to flickering [10]. They have been explored broadly for different potential applications, counting fluorescence organic names, photovoltaic cells, light-emitting diodes, and optical sensors.

In this submitted work, it was endeavored to utilize NPs of the fluorometric gold mineral without the use of natural and inorganic stabilizers [11]. Appropriately, brilliant NPs would be connected within the advancement and consider a more exact and less run-time in mistake proportion and a moderately brief and particular strategy for deciding CEF in pharmacological arrangements and organic liquids (human serum and urine samples). Color unearthly methods are still utilized within the examination of numerous drugs since they take a lower toll compared to other strategies of investigation, effortlessness, and common sense. Here we report an unused colorimetric strategy for recognizing CEF in immaculate and pharmaceutical dosage forms. The spectroscopic strategy was based on the collection of gold nanoparticles when including sedate. Following an examination of the response conditions, the results showed an excellent retention range and transmission capacity consistent with the type of medication. Degree of collection of brilliant NPs changed the color from ruddy to purple to dim blue, depending on the estimate of the brilliant NPs. In other words, brilliant NPs can recognize compounds that don't contain chromophore bunches or color-derived subordinates that can be troublesome to synthesize. Since these physico-chemical properties intriguing to numerous analysts, brilliant NPs have wide areas of application in different areas of chemistry [12-15]. It can be accepted that this current technique is capable of being used in preclinical pharmacokinetic studies in CEF.

 

EXPERIMENTAL

 Instrumentation

 All fluorescent spectroscopy of the drugs under study was performed using the SPEX Fluorolog-2 Spectrofluorometer scale. The 450W xenon lamp was used as an excitation light source for the spectrum of the R928 multiplier tube that operates at 950 volts (Hamamatsu) as a detector. Fissure and excitation, the increase time, and the integration time were set to 1.25 mm, 1 nm, and 1 second, respectively. All spectral data was analyzed by SPEX DM 3000F spectroscopy.

A single cell-holder, the JENWAY 6715 UV/visible spectrophotometer equipped with 10-mm matched quartz cells, was employed for all absorbance measurements.

 

Materials and reagents

All solvents and reagents used were of the highest purity: Chloroauric acid (HAuCl4), obtained from Fischer Chemical, Fischer Scientific UK Limited, U.K. Sodium citrate. Acetate buffer pH = 5: Dissolve 13.6 g of sodium acetate and 6 mL of glacial acetic acid in sufficient water to produce 1000 mL of acetate buffer [16].

 

Authentic and Pharmaceutical preparation:

Ceftobiprole sodium with purity 99.11 [6] and Zeftera® Vials, each vial containing 1 g ceftobiprole sodium per vial with batch no. 209467, Basilea Co., Switzerland.

Standard solutions

Solutions of 100 µg/mL CEF were prepared by dissolving 10 mg of pure drug in a 100 mL volumetric flask with distilled water, which was then completed to 100 mL with distilled water and further diluted to 10 µg/ml.

 

General Procedure

Procedure for preparation of citrate-stabilized gold nanoparticles

The nitrogen and zinc particles required by the coatings were arranged by using reduced chloroauric acid with sodium citrate [17]. In a 150-mL glass, 2.0 mL of 1% HAuCl4 and 90 mL of water were combined and heated to 95 °C. 5 ml of a 1% sodium citrate solution was added and stirred continuously. The solution was held at 95 degrees Celsius for 10 minutes.

When the arrangement changes its color to light ruddy, it is cooled to room temperature and exchanged into a 100-mL volumetric flask. Au NPs are kept at 4 °C until utilized.

 

Procedure for determination of the cited drug spectrofluorimetrically

All glassware was washed with nitric acid and then removed with distilled water. NPs were utilized as a fluorescent test to determine CEF. The taking-after method has been embraced. Fitting amounts of NPs and both investigation and application medicate were included in the 4 ml FL cell. The arrangement is weakened with refined water and blended, after which the thickness of FL is measured. The excitation wavelength was set at 452 nm, and the outflow wavelength was at 585 nm.

 

Procedure for determination of the cited drug spectrophotometrically

All glassware sets utilized within the taking after strategies were cleaned by aqua regia, washed altogether in double-deionized water, and dried in discuss earlier to utilize. In a 5 mL volumetric flask, diverse aliquots of CEF were put; at that point, suitable volumes of acetic acid derivation buffer and Au NP arrangement were included, completed to 5 mL with distilled water, and let stand at room temperature for 5 min. Absorbance was measured at the appropriate ʎ max against the reagent, clear-treated additionally.

 

 

Figure 1. Chemical structure of CEF

 

Assay of pharmaceutical preparation

In a 100-ml volumetric flask, a particular amount of powder was broken down to 10 mg of immaculate pharmaceutical, which was then stamped with distilled water. At that point, it was completed to 10 µg / mL and steps were completed as common steps were connected.

 

Assay of biological fluids

For human plasma

One-milliliter aliquots of plasma were distributed into two centrifuge tubes. The plasma was spiked with aliquot concentrations of the studied drug in stock solution. The tubes were blended well by employing a vortex blender. The tubes were deproteinized twice with acetonitrile. The centrifugation was done for 20 minutes at 7000 rpm. The centrifuges were transferred to unused and clean centrifuge tubes, where they evaporated. The buildups were changed to methanol and transferred to 5 mL volumetric flasks, and the volumes were adjusted to the line with distilled water. Aliquots of 2 mL from each arrangement were transferred to a 25 mL volumetric flask, the specified volumes of CEF and buffer were taken, and the volume was completed to the line with distilled water. The procedure was completed as described in pharmaceutical dosage forms.

For urine samples:

1.0 mL of urine and 2.5 mL of standard CEF arrangements (100 µgmL-1) were blended. The blend was centrifuged by altering the centrifuge machine at 3000 rpm for a time period of 10 minutes. Clear fluid was poured into a 50-mL volume flask, and the volume was completed with distilled water. Appropriate volumes (0.2, 0.4, and 0.6 µgmL-1) from this arrangement were examined utilizing the general method depicted over.

 

RESULTS AND DISCUSSION

Examining NPs utilizing both silver and gold is imperative because of their interesting optical properties. Brilliant NPs influence assimilation due to surface plasmon reverberation (SPR), which happens when the electron field vacillates around the NP due to the assimilation of light vitality. It depends on the measure of the NP; in this way, the status of the collection of brilliant NPs has an impact on optical retention. NPs were introduced from gold by expelling it using sodium citrate. NPs synthesized a well-known retention band at 520 nm. When CEF was included, the most extreme retention showed up due to the amassing of fluorometric spectral gold NPs (Figure 2) and the optical range (Figure 3). NPS was utilized in the investigation and assessment of CEF spectroscopy and its ghostly measurements.

 

Characterization of gold nanoparticles

Synthesis of gold nanoparticles

Brilliant NPs were made as a result of particular steps: constraining the arrangement of chloroauric acid in Au atoms, and these atoms were expected to create an Au mass; expanding their estimate to a particular examined volume; and these clusters were examined [14]. The relevant scheme 1 illustrates the composition of the golden NPs [15].

Reduction of chloroauric acid

Golden NPs were synthesized by reducing the volume of tetrachloroauric acid after removing sodium citrate according to the following equations [15]:

AuCl3 + C6H5O73- → AuCl + SADC + CO2 + H+ + 2Cl-

3AuCl → 2Auo + AuCl3

Gold was dissolved in the form of Au3+. When the reducing agent was added, the gold granules were studied and monitored until the solution exceeded saturation in concentration. Then the particles overlapped and entwined in a process called nucleation. The rest of the gold granules are linked to the nucleation and growth sites [18].

 

 

 

 

 

Figure 2. FL emission spectra (excitation at 450 nm) of CEF-gold NPs at 585 nm

 

 

 

Figure 3. Absorption spectra of the reaction between Au NPs and CEF

 

 

Stabilization of nanoclusters against aggregation

Two sorts of steadiness were examined: electrostatic stabilization for stabilization [17], Figure 4. In this strategy, we depended on electrostatic steadiness, utilizing sodium citrate, which acts as a diluent and stabilizer. The adjustment of the two strengths protected the properties of the components, that's, the scornful drive of the two-fold layer and the appealing constraint of Vanker Untrue [19].

Effect of sodium citrate concentration

The concentration of sodium citrate increases the soundness of the brilliant superposition with the lessening handle. In this composition, distinctive concentrations are considered to get the most excellent non-superoxide NPs. Another 10, 20, and 40 mM NaCl concentration was utilized. The arrangement utilizing sodium citrate 10 and 20 mm appeared to get together marks by giving blue, but a 40 mm arrangement was steady, which gave a clear, relentless ruddy color. A switch arrangement with a low concentration of citrate can result in the authoritative of NPs to each other within the arrangement due to their expanded volume, whereas the use of a better concentration of sodium citrate decreases the measure of the AuNPs and shapes well-partitioned atoms. With respect to the quality of the NPs, it was found that the best concentration was 40 mm [18].

 

 

 

Spectrofluorimetric method

The concentration of brilliant NPs essentially influenced the degree of fluorescence (FL) of the pharmacokinetic framework. The thickness of FL expanded and reached its highest value, when the concentration of nanoparticles was within the range of 4.0 x 10-5 to 6.0 x 10-5 mol L-1. The extra increment in the center caused the FL to escalate and diminish. Given the precision of the affectability and straight extend, the use of 6.0 x 10-5 mol L-1 was accomplished as a perfect concentration for NPs.

 

Fluorescence (FL) spectral characteristics

Figure 2 shows the emission of the brilliant NP spectra in the presence of CEF. Brilliant NPs appeared with a particular fluorescence spectra of emanation at 585 nm and excitation at 452 nm. It was noted from the experiment that the results were suppressed by the intensity of FL in golden NPs without any change after adding CEF to the steps of the experiment. This cooling may be the result of the arrangement of a non-fluorescent ground condition complex between the considered drugs and the indicator. The non-fluorescent compound retains light and promptly returns to the ground state without outflow or photons, as a result of which, as it were uncomplicated for giving FL emission, in this manner quenching the whole FL escalated.

The effect of cooling can be described by a strict volumetric [20]:

 (f0 / f) - 1 = KCQ 

Where F0 is the FL intensity of absorption from the SIL quenched, f is the density of FL at the Q concentration of the CEF, and K is a proportional constant. If the system follows the law, the f0 / f plot versus CQ leads to a single difference on the y axis and a slope equal to K. The fo / f line increases with an increase in the amount of CEF. The effect of rapid cooling with respect to temperature was lower at a higher temperature. Dynamic cooling depends on diffusion. Since high temperatures result in wider diffusion coefficients, after study we expect the cooling constants to increase with increasing temperatures. Therefore, it can be concluded that the cooling mechanism is stable and constant by CEF due to the formation of a non-fluorescent compound between the golden NPs and the CEF.

 

Spectrophotometric method

Optimization of reaction conditions

To obtain the best results and concentrations of CEF under study to determine the best optical spectra using golden nanocomposites, the following has been studied:

Effect of buffer pH

The pH influences the solidity of the interaction between Au NPs and the drugs under examination. The pH extend was tested on the ice stability of NPs 2-10 utilizing diverse pH media (acetic acid derivation dielectric, phosphate dielectric, chloride dielectric, and borate dielectric). It was found that in the case of less than 5, brilliant NPs would collect more due to citations of positive charges [15]. Whereas above 6, a decrease within the ghastly estimation was found, and after that, the acetic acid derivation dielectric was chosen at pH 5 to deliver a steady arrangement and a particular concentration of CEF (Figure 5).

Effect of volume of gold nanoparticles solution

The volume of AuNPs was examined, and it was found that the increment was up to 1.5 mL. It can be said that the ghostly retention would diminish in the event that the AuNPs were not found within the rectifying condition for these values. This may be ascribed to less authoritative items, which made the escalated less; on the contrary, over the top gold NPs would connect with the drugs beneath think about comparatively, so the number of drugs beneath think about altering the brilliant NPs would diminish, coming about in diminished unearthly concentrated (Figure 6 ).

Effect of temperature

It was found that the heat has no impact. The tests were carried out at diverse temperatures within the range of 25, 40, 60, and 80 ° C. No change was observed within the ghostly state of the superpositions; appropriately, the tests were performed at room temperature (Figure 7).

Effect of time

The period analyzed indicates that 5 minutes is the optimal time to achieve the greatest thickness of medication within consideration (Figure 8).

Order of addition

In tests, distinctive components were examined. The most appropriate groups were buffer, dielectric acetic acid derivation, and then brilliant NPs; the expansion of this medication to brilliant NPs revealed a monitored rate of superposition arrangement.

 

Method validation

Linearity

The bend was considered to determine the CEF beneath ponder within the recommended way by drawing FL or ghostly assimilation signals versus sedate concentrations. Brew code plots were little direct interceptors with great relationship coefficients showing great linearity over work center ranges. Molar absorptivity, relative standard deviation, explanatory standard mistake, location, and estimation limits were overestimated (Table 1).

Sensitivity

The least distinguishable analyzer (LOD) concentration is considered, but not fundamentally quantitatively as the desired esteem is. The quantitative restraint (LOQ) was the most reduced concentration of the analyzer according to the laws, which can be measured with fitting exactness. LOD and LOQ were assessed utilizing the following conditions concurring with ICH rules [18]: LOD = 3.3 ᾳ / S and LOQ = 10 ᾳ / S, where ᾳ is the standard deviation of rehashed clear reactions (below the same conditions for analyzing the drug under study, and S may be an incline calibration bend): LODs and LOQs are calculated in Table 1.

Accuracy 

The strategies of examination of CEF beneath consider were examined in terms of accuracy with the distributed strategy for the same sedate beneath consider. The result of the factual examination of the outcomes gotten by the proposed and comparative strategies of the examined drugs appeared to be that the calculated values did not surpass the comparative hypothetical values, which demonstrated that there were no factual contrasts between the proposed strategies and the comparison strategies of the drugs under consideration. A factual comparison [6] of the results was performed, counting a T-test and understudy differentiate proportion test at a 95% certainty level (Table 2).

Figure 4. Types of nanoparticles stabilization	Figure 5. Effect of pH on aggregation rate in presence of CEF

 

 

 

 

 

Figure 6. Effect of volume of gold nanoparticles on aggregation rate in presence of CEF

 

 

Figure 7. Effect of temperature on aggregation rate in presence of CEF

 

 

Figure 8. Effect of time on aggregation rate in the presence of CEF

 

 

Table 1. Analytical parameters for spectrofluorimetric and spectrophotometric determination of CEF using gold nanoparticles

Spectrofluorometry
λex/em (nm)	450/585
Volume of gold (mol L-1)	4x10-5
Volume of buffer (mL)	0.1
Time (min)	5
Temperature (oC)	25
Linearity range (ng mL-1)	20-120
LOD (ng mL-1)	5.54
LOQ (ng mL-1)	18.78
Slope (b)	1.976
Intercept (a)	5.5422
Correlation coefficient (r)	0.9989
Spectrophotometry
Λmax (nm)	625
Volume of gold (mol L-1)	1.5
Volume of buffer (mL)	0.1
Time (min)	5
Temperature (oC)	25
Linearity range (µg mL-1)	0.2-1.0
LOD (µg mL-1)	0.06
LOQ (µg mL-1)	0.14
Slope (b)	0.898
Intercept (a)	0.073
Correlation coefficient (r)	0.9999

 

 

Table 2. Determination of CEF through gold nanoparticles formation

Statistics
Spectrofluorometry
Mean (X-)	99.91
Number of experiments (N)	6
Variance (V)	0.61
Standard deviation (±SD)	0.77
Relative standard deviation (%RSD)	0.77
Spectrophotometry
Mean (X-)	100.11
Number of experiments (N)	6
Variance (V)	0.31
Standard deviation (±SD)	0.61
Relative standard deviation (RSD%)	0.61

 

Intra-day precision (repeatability)

Three distinctive concentrations of sedate beneath consider (inside direct ranges) were arranged for CEF in its crude shape and analyzed within the way proposed three continuous times on the same day. The values of the relative standard deviation and the relative mistakes (E %) of the strategy created were calculated utilizing the following equation:

Er % = (existing - included) / included x 100

Inter-day precision (intermediate)

Three repeated trials counting three diverse concentrations were considered in one day (inside the straight condition ranges) of the medication below utilizing the proposed strategy over a period of three days. The relative deviation of the ghostly estimation and the rate of relative errors (Er %) were calculated.

Selectivity

The interference obligations of the methods under study were implemented to selectively explore the effect of common excipients that could be added during the combinations. Under the conditions of novel experiments, to a specific concentration of CEF under study, common excipients of lactose, sodium dodecyl sulfate, starch, and magnesium stearate were added and analyzed.

Robustness

 Strength is the degree of the capacity of the ghastly strategies beneath ponder to outlive without being affected by slight changes within the contrasts within the strategies beneath consider. Assessment steps were taken by making little progressive changes in one parameter, whereas the others remained unaltered, such as the estimate of the standard gold arrangement and the estimate of the buffer acetic acid derivation. The impact of the changes on the ghostly assimilation was considered by calculating the recuperation of ± RSD%, and the changes had an exceptionally small impact on the effectiveness of the strategies created, giving a sign of the wellbeing and exactness of the strategy proposed amid its research facility application for the examination of the sedate inspected.

 

Analytical application

Determination of the studied drug in its pharmaceutical preparation

The validity of the proposed FL and spectrophotometric techniques for the determination of CEF in pharmaceutical preparation was investigated. After sample preparation and adequate dilution, the method was applied to the direct determination of CEF in its dosage form using a calibration curve. The summarized results for the analysis of pharmaceutical dosage forms are shown in Table 3. The results showed that the proposed FL method could be applied successfully for the determination of CEF in its dosage form. A student's t-test of the proposed and other strategies [6] in terms of exactness and accuracy and an F-test for variance ratio were used to investigate the materials using both the proposed and comparative strategies. Results showed that there was no noteworthy distinction between the proposed and other strategies in terms of exactness and accuracy.

Determination of the studied drug in a spiked serum sample

The possibility of applying the FL strategy to human serum tests has been examined. A strategy of examination created to determine CEF was connected in a serrated test after the sedimentation of proteins and inside materials from serum tests for evacuation by including an acetonitrile centrifuge at 4000 rpm. At that point, the supernatants were taken and weakened with refined water. Backed serum tests expanded solidity with standard CEF to affirm crest position and compare the FL reaction to the extra concentration. Compatibility with straight conditions was monitored by including the considered documentation in the already braced serum test, which affirmed the appropriateness of the proposed strategy to human serum and gave comparative results in exactness and recurrence in Table 4.

Determination of the studied drug in a urine sample

The pertinence of the examined strategy was approved to distinguish CEF in opening pee. It was found that a coordinated distinguishing proof of CEF in pee is conceivable through exceedingly weakened tests. The unused steps were performed on pee tests and rehashed five times for each test. The reasons are given in Table 4. From the recuperation information, it was observed that CEF can be characterized lattice included with the same comes about in exactness and recurrence (Table 4).

 

 

Table 3. Application of the proposed methods for determination of CEF in Zeftera® Vials

Statistics
Spectrofluorometry
Mean (X-)	98.21
Number of experiments (N)	3
Variance (V)	1.82
Standard deviation (±SD)	1.41
Relative standard deviation (RSD%)	1.38
Spectrophotometry
Mean (X-)	99.06
Number of experiments (N)	3
Variance (V)	1.00
Standard deviation (±SD)	1.00
Relative standard deviation (RSD%)	0.99

 

Table 4. Application of the proposed spectrofluorimetric method for determination of CEF in serum and urine samples

Statistics
Serum Sample
Mean (X-)	97.99
Number of experiments (N)	3
Variance (V)	4.14
Standard deviation (±SD)	2.10
Relative standard deviation (RSD%)	1.94
Urine Sample
Mean (X-)	100.98
Number of experiments (N)	3
Variance (V)	3.47
Standard deviation (±SD)	1.98
Relative standard deviation (RSD%)	1.98

 

 

 

CONCLUSION

The authors utilized one of the well-established properties of gold NPs, conglomeration, for spectrofluorimetric and spectrophotometric assurance of CEF. The novel fluorimetric strategy based on the FL extinguishing of gold NPs has points of interest in terms of effortlessness, rate, and affectability. The created strategy was effectively connected to the assurance of the considered medications as an FL test in human serum and urine. The exploratory findings propose that gold NPs may provide a modern course of fluorophore for use in chemical detection and biomedical applications. Too, gold NPs tended to total through electrostatic interaction with the cited cationic medicate, which brought about the arrangement of an unused ruddy-moved band. The information given already uncovered that the proposed strategy was straightforward, delicate, reasonable, and effectively appropriate to the examination of drugs in their pharmaceutical measurement shapes with great precision and exactness.

Author Contributions:  All authors contributed to the study's conception and design. Data collection and analysis were performed by all authors. The first draft of the manuscript was written by Hesham Salem, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Author Declarations: Ethics Approval the Collection of samples was authorized by the Ethics Committee of Minia Hospital.

Conflicts of Interest/Competing Interests: The authors declare that they have no conflict of interest.

 

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