Determination Of Penicillin Research Articles

Dependence of the Group Specificity of Immunoenzyme Determination of Penicillins in Milk on the Temperature and Duration of Antibiotic Cross Reactions with Polyclonal Antibodies

Dependence of the Group Specificity of Immunoenzyme Determination of Penicillins in Milk on the Temperature and Duration of Antibiotic Cross Reactions with Polyclonal Antibodies

Early testing and grade of the reaction are affecting factors of skin test positivity in β-Lactam allergies.

Background: β-Lactams are the most widely used antibiotic family in the world. Nevertheless, they also stand out as the primary culprits for inducing drug hypersensitivity reactions (HSR). Methods: Between May 2018 and March 2023, patients with suspected HSRs to β-lactams, who underwent skin tests (ST), were retrospectively screened. The determinants of allergenic penicillin (DAP) tests, which include penicillin minor and major determinants, clavulanic acid, and amoxicillin, along with ampicillin, sulbactam, the identified culprit drugs, and alternative cephalosporins, which include cefuroxime, ceftriaxone prick and/or intradermal tests, were administered. The analysis focused on identifying positive ST results and determining the true HSRs rates in this patient cohort. Results: Of the 147 patients, 78.9% (n = 116) were women and the median (minimum-maximum) age was 41 years (18-71 years). Mild HSRs (grades 1-2) were observed in 72.78% (n = 107), whereas 24.4% (n = 36) had severe reactions (grades 3-4) and 2.7% (n = 4) had an unknown grade. Of the patients, 64% (n = 94) experienced HSRs within the first hour after the last dose of the identified culprit drug. The overall positivity rate for all STs was 26.5% (n = 39). ST positivity rates were notably higher in individuals who had experienced HSRs within the past 6 months (p = 0.02) and those with severe anaphylaxis (p < 0.001). Conclusion: β-Lactam ST positivity is higher, especially in those with grades 3-4 reactions and consulted a physician within the first 6 months after their HSRs.

Determination and Identification of Antibiotic Residues in Fruits Using Liquid Chromatography-High-Resolution Mass Spectrometry (LC-HRMS).

Antibiotic residues may be present in fruit products from trees that were treated to combat bacterial diseases such as citrus greening or blight. A liquid chromatography-high-resolution mass spectrometry (LC-HRMS) method was developed for the simultaneous determination and identification of streptomycin, kasugamycin, penicillin, and oxytetracycline residues in fruit. Samples were extracted with acidic methanol and separation was optimized for a hydrophilic interaction LC column. A Q-Exactive HRMS instrument was used to obtain product ion spectra for analyte identification. Quantitation was performed with matrix-extracted calibration curves and internal standard correction. The method was tested on many different types of fruit. In general, fortified samples demonstrated acceptable recoveries (82-116%) and reproducibility (<15% RSD). Method detection limits for these analytes were well below the established US EPA tolerance levels. It was also possible to analyze the fruit extracts prepared using this method for additional chemical contaminants using LC-HRMS.

A penicillinase-modified poly(N-isopropylacrylamide-co-acrylamide) smart hydrogel biosensor with superior recyclability for sensitive and colorimetric detection of penicillin G

Antibiotics are widely used for treating bacterial infections. However, excessive or improper use of antibiotics can pose a serious threat to human health and water environments, and thus, developing cost-effective, portable and effective strategies to analyze and detect antibiotics is highly desired. Herein, we reported a responsive photonic hydrogel (RPH)-based optical biosensor (PPNAH) with superior recyclability for sensitive and colorimetric determination of a typical β-lactam antibiotic penicillin G (PG) in water. This sensor was composed of poly(N-isopropylacrylamide-co-acrylamide) smart hydrogel with incorporated penicillinase and Fe3O4@SiO2 colloidal photonic crystals (CPCs). The sensor could translate PG concentration signals into changes in the diffraction wavelength and structural color of the hydrogel. It possessed high sensitivity and selectivity to PG and excellent detection performances for other two typical β-lactam antibiotics. Most importantly, due to the unique thermosensitivity of the poly(N-isopropylacrylamide) moieties in the hydrogel, the PG-responded PPNAH sensor could be facilely regenerated via a simple physical method at least fifty times while without compromising its response performance. Besides, our sensor was suitable for monitoring the PG-contaminated environmental water and displayed satisfactory detection performances. Such a sensor possessed obvious advantages of superior recyclability, highly chemical stability, low production cost, easy fabrication, wide range of visual detection, simple and intuitive operation for PG detection, and environmental-friendliness, which holds great potential in sensitive and colorimetric detection of the PG residues in polluted water.

The Dependence of the Group Specificity of Immunoenzyme Determination of Penicillins in Milk on the Temperature and Duration of Antibiotic Cross Reactions with Polyclonal Antibodies

The Dependence of the Group Specificity of Immunoenzyme Determination of Penicillins in Milk on the Temperature and Duration of Antibiotic Cross Reactions with Polyclonal Antibodies

New dual-modal electrochemical and photoelectrochemical aptasensing strategy for the sensitive detection of penicillin G based on an iron-doped covalent triazine framework

The sensitive determination of penicillin G (PG) in complex environments is essential for guaranteeing food safety and human health. Herein, a new dual-modal electrochemical (EC)–photoelectrochemical (PEC) aptasensing strategy has been proposed based on an Fe-doped covalent triazine framework (denoted as Fe-MA-Dha-CTF) synthesized by using 2,5-dihydroxyterephthalaldehyde (Dha) and melamine (MA) as building blocks and utilized to detect trace PG. Compared with the platform based on metal-free MA-Dha-CTF, the constructed Fe-MA-Dha-CTF demonstrated enhanced photoelectric conversion efficiency, superior electrochemical activity, and high bioaffinity toward the PG-targeted aptamer. The established Fe-MA-Dha-CTF-based dual-modal EC–PEC aptasensor exhibited ultralow limits of detection of 0.08 and 0.7 fg mL−1 as deduced by using EC and PEC techniques, respectively, within a wide PG concentration range of 1 fg mL−1 to 10 ng mL−1. Moreover, the proposed EC–PEC aptasensor exhibited high selectivity, good stability, acceptable reproducibility, and great practicability in diverse real samples. The CTF-based dual-modal aptasensing approach presented here can extend the application of porous-organic frameworks in biosensing.

Molecular docking of human serum albumin with penicillin G determinants

Background: Human serum albumin (HSA) is the main pharmacokinetic effector of many medications, including penicillin G and its metabolites. An urgent problem of practical medicine is immediate hypersensitivity reactions caused by penicillin toxicity (about 8 % compared to other medications), accompanied by skin pathology, anaphylaxis, and fatality. Objectives: The aim of this study is to describe the structures of penicillin G determinants-HSA complexes and to identify favorable binding sites and the amino acid residues involved in the interaction. Material and Methods: The crystal structure of HSA (ID: 1AO6 from Protein Data Bank) (www.rcsb.org) was chosen as a docking target. Molecular docking methods (AutoDock Tools 1.5.7, AutoDock Vina 1.1.2) were used to gain insight into the interaction of HSA with the major (benzyl penicilloyl G, penicillanic acid) and minor (penicillamine, penicilloic acid, penilloic acid) determinants of penicillin G. Visualization of docking results was implemented in PyMol 2.5. The Protein Plus server (https://proteins.plus) was used to evaluate potential binding pockets. The PLIP tool (https://plip-tool.biotec.tu-dresden.de) was used to identify non-covalent interactions between HSA and its ligands. Results: The molecular docking data indicate that the major determinants of penicillin G are involved in the formation of hydrogen bonds with such HSA residues as Trp214, Arg218, His242, and Asn295; for the minor determinants — Asp108, His146, Tyr148, Ser193, Arg197, Gln204. Both types of determinants are located in the hydrophobic cavity of subdomains IIA and IB. Hydrophobic interactions are present mainly between penicillin G determinants and amino acid residues of subdomain IIIA, such as Ala350, Asp451, Tyr452, and Gln459. Conclusion: The study of penicillin G determinants-HSA complexes is important in the pathogenesis of antibiotic allergy. Identification of specific binding sites can be useful for the development and synthesis of new immunogenic antigens (complexes of major and minor determinants of penicillin G with HSA) that can stimulate the immune system and produce specific antibodies to prevent allergic reactions.

Use of Magnetic Carbon Nanocomposites in the Formation of a Recognition Layer of a Piezoelectric Immunosensor for the Determination of Penicillin G

Conditions for the formation of a recognition layer of a piezoelectric immunosensor based on magnetic carbon nanocomposites (MCNCs) under the action of an external magnetic field are studied. The effects of the size and number of magnetic nanoparticles (MNPs) in the composite on the analytical characteristics of the gravimetric immunosensor are revealed. Scanning electron microscopy is used to determine the average sizes of Fe3O4 magnetic nanoparticles synthesized by coprecipitation. It is noted that the minimum weight and stability of the recognition layer were observed for the nanocomposite obtained at a ratio of carbon nanotubes and MNPs with an average diameter of 22 nm equal to 3 : 1. The formation of peptide bonds between the MCNCs and a penicillin G conjugate was established by IR spectrometry. It was shown that the use of magnetic carbon nanocomposites in the formation of a recognition layer makes it possible to significantly simplify the procedure for preparing a piezoelectric sensor for analysis and reduce its duration from 24 to 1.5 h. The range of the determined antibiotic concentrations is 1–450 ng/mL, the limit of detection is 0.5 ng/mL.

Development of a Dispersive Liquid-Liquid Microextraction for the Spectrophotometric Determination of Penicillin G in Milk and Biological Samples

Development of a Dispersive Liquid-Liquid Microextraction for the Spectrophotometric Determination of Penicillin G in Milk and Biological Samples

A novel triple templates molecularly imprinted biosensor assisted by first derivatives of second-order hydrodynamic differential normal pulse voltametric data and multi-way calibration methods for simultaneous biosensing of penicillin, tetracycline and amoxicillin in dairy products: A novel multi-disciplinary study

In this work, a screen printed carbon electrode (SPCE) was used as a platform to fabricate a novel electrochemical biosensor assisted by the first derivatives of the second-order hydrodynamic differential normal pulse voltammetric (HDNPV) data modeled by multivariate curve resolution alternating least squares (MCR-ALS) and parallel factor analysis applied to shift invariant amplitude spectra (PARASIAS) for simultaneous determination of penicillin (PC), tetracycline (TC), and amoxicillin (AC) in dairy products. The SPCE was modified with poly(3,4-ethylenedioxythiophene) (PEDOT) by electropolymerization of the EDOT. Triple templates molecularly imprinted polymers (TTMIPs) were synthesized by using acrylic acid as the functional monomer, and TC, PC, and AC as the template molecules, and epichlorohydrin as a crosslinking agent. The TTMIPs were mixed with multi-walled carbon nanotubes-graphene-ionic liquid (MWCNTs-GR-IL), and drop-casted onto the surface of PEDOT/SPCE. The first derivatives of individual HDNPV responses of the TTMIPs-MWCNTs-GR-IL/PEDOT/SPCE to TC, PC and AC were univariately calibrated, and their linear ranges were used to develop second-order calibration models using MCR-ALS and PARASIAS whose performances were validated by application of them to a validation set with the aim of choosing the best model to assist the biosensor for the analysis of yogurt, milk, and yogurt drink as real samples. The biosensor assisted by MCR-ALS showed a better performance than PARASIAS which forced us to make a final decision on choosing it to assist the biosensor for the analysis of real matrices. The methodology developed in this work was benefited from TTMIIPs, good structure of the biosensor, and three-way calibration models which helped us to develop a novel electroanalytical method for simultaneous determination of TC (0.1–9 pM), PC (1–10 pM), and AC (0.35–8 pM) whose performance in analysing real matrices was comparable with the HPLC-UV as the reference method.

Clavulanic Acid Is a Leading Culprit Beta-Lactam in Immediate Allergic Reactions to Penicillins.

Clavulanate, a beta-lactam associated with amoxicillin, is frequently prescribed in patients at all ages. Recent data implicate amoxicillin-clavulanate in up to 80% of beta-lactam allergy cases. We assessed clavulanate's role in inducing allergic reactions to this combination treatment, with a focus on selective immediate reactions. Adults (≥ 16 years) reporting a history of immediate reactions to amoxicillin-clavulanate were evaluated through a beta-lactam allergological workup, using modified European Academy of Allergy and Clinical Immunology guidelines. Patients first underwent skin testing, and if negative, drug provocation tests. Expected outcomes were: Group A, subjects with immediate reaction to classical penicillin group determinants (penicilloyl polylysine, minor determinants mixture, and/or penicillin G); Group B, subjects with selective immediate reaction to amoxicillin; Group C, subjects with selective immediate reaction to clavulanate and Group D, those immediate reactions with co-sensitization to clavulanate plus penicillin group determinants or amoxicillin. Of 1,170 included patients, 104 had immediate reactions: 36.5% to penicillin group determinants (Group A), 26.9% to amoxicillin (Group B), 32.7% to clavulanate (Group C), and 3.8% to clavulanate plus penicillin determinants or amoxicillin (Group D). Diagnosis was made by skin testing in 79%, 75% and 47% of the patients, respectively, in the first 3 groups (P < 0.001). Drug provocation tests were necessary to establish most other diagnoses. Anaphylaxis predominated over urticaria/angioedema in all groups. Selective immediate reactions to clavulanate accounted for over a third of cases with confirmed reactions after amoxicillin-clavulanate intake, with more than half experiencing anaphylaxis. Within this group, skin test sensitivity was below 50%. People taking amoxicillin-clavulanate may also be co-sensitized to both drugs.

Multiresidue chromatographic method for the determination of antibiotic residues in honey by high-performance liquid chromatography with DAD detection

Clandestinely, consumers may be exposed to antibiotic (ATB) residues in honey, which could pose a health concern. For the first time, the simultaneous determination of Florfenicol (FF), Penicillin G (PG), and Tetracycline is described in this paper. The multiresidual method was developed and optimized using high-performance liquid chromatography (HPLC) coupled to a diode array detector (DAD). These ATBs were separated on a C18 analytical column after a cleanup process followed by solid-phase extraction (SPE). For the first time, the chromatographic conditions were perfected. After the method validation process, the method was used to assess ATB residues in four Lebanese honey samples. ATBs were separated in less than 15 min with an isocratic elution using a mixture of 80 % potassium dihydrogen phosphate aqueous solution, and 20 % acetonitrile. The UV detection was performed at 350 nm for TC, 224 nm for FF, and 230 nm for PG. The proposed method was linear (R2 ≥ 0.996) within the concentration ranges of 0.7-17.5 mg.Kg 1 for the three compounds. Both intra- and inter-day precision, expressed as RSD, were ≤15 %. The method was subsequently successfully applied to analyze examined ATB residues in honey samples collected from Lebanese beekeeping. The method described could be a valuable tool to conduct a comprehensive survey of honey samples produced in Lebanon, especially in the lack of serious national oversight.

Molecularly imprinted electrochemical based on NiO nanostructured modified glassy carbon electrode for the electrochemical determination of penicillin in urine of pregnant women infected with Group B Streptococcal

Molecularly imprinted electrochemical based on NiO nanostructured modified glassy carbon electrode for the electrochemical determination of penicillin in urine of pregnant women infected with Group B Streptococcal

Pre-column derivatization with trimethylsilyl diazomethane coupled with ASE-SPE-GC-MS/MS method for the quantification and validation of penicillin G residues in poultry tissues and pork

A novel, simple and high-sensitivity gas chromatography-tandem mass spectrometry (GC-MS/MS) method combined with external calibration was developed for the qualitative and quantitative determination of penicillin G residues in poultry tissues and pork. Samples were extracted by an ASE 350 apparatus with 0.2 mol/L phosphate buffer solution (pH 8.0) and purified on HLB solid-phase extraction (SPE) cartridges (60 mg/3 mL). Trimethylsilyl diazomethane (TMSD) was selected as the derivatization reagent for precolumn derivatization. The recoveries of analytes at the limit of quantitation (LOQ) and 25, 50 and 100 μg/kg ranged from 80.67% to 96.18%, with RSDs below 3.17%. The determination coefficient (R2) values were above 0.9993 in the corresponding concentration range. The limits of detection (LODs) (S/N ≥ 3) and LOQs (S/N ≥ 10) ranged from 1.50 to 4.10 μg/kg and 4.50–8.20 μg/kg, respectively. In addition, the newly established method was implemented to analyze poultry tissues and pork purchased from a local market, demonstrating the sensitivity and applicability of the method.

A new voltammetric sensor for penicillin G using poly(3-methylthiophene)-citric acid modified glassy carbon electrode

In this study, an effective voltammetric sensor was developed for determination of penicillin G by electropolymerization of 3-methylthiophene with citric acid on glassy carbon electrode. Penicillin G selective electrode was prepared in water:acetonitrile (1:1) mixture containing 0.1 M sodium perchlorate as electrolyte medium with cyclic voltammetry. Then, the electrochemical behavior of penicillin G on poly(3-methylthiophene) based membrane modified electrode was investigated by cyclic voltammetry, differential pulse voltammetry and differential pulse stripping voltammetry techniques. When the penicillin G responses of the modified glassy carbon electrode were compared with the bare glassy carbon electrode responses, it was seen that the selectivity and sensitivity of the penicillin G responses of the modified electrode significantly increased. A linear calibration graph for penicillin G was obtained in the concentration range of 0.07 to 4.0 mM with the prepared electrode. The R2 value was calculated as 0.9999 from the linear calibration curve. The limit of detection and limit of quantitation of the poly(3-methylthiophene)-citric acid modified glassy carbon electrode for penicillin G were calculated as 8 μM and 28 μM, respectively. In addition, the developed electrode for the detection of PeG was applied to milk samples. The recovery efficiency of the poly(3-methylthiophene)-citric acid modified glassy carbon electrode was obtained in the range of 93 % and 103 % for three different milk samples. As a result; the modified electrode can be used for residue penicillin detection in food and biological samples.

Changes in Sensitization Patterns in the Last 25 Years in 619 Patients with Confirmed Diagnoses of Immediate Hypersensitivity Reactions to Beta-Lactams.

Beta-lactam (BL) drugs are the antibiotics most prescribed worldwide due to their broad spectrum of action. They are also the most frequently implied in hypersensitivity reactions with a known specific immunological mechanism. Since the commercialization of benzylpenicillin, allergic reactions have been described; over the years, other new BL drugs provided alternative treatments to penicillin, and amoxicillin is now the most prescribed BL in Europe. Diagnosis of BL allergy is mainly based on skin tests and drug provocation tests, defining different sensitization patterns or phenotypes. In this study, we evaluated 619 patients with a confirmed diagnosis of BL-immediate allergy during the last 25 years, using the same diagnostic procedures with minor adaptations to the successive guidelines. The initial eliciting drug was benzylpenicillin, which changed to amoxicillin with or without clavulanic acid and cephalosporins in recent years. In skin tests, we found a decrease in sensitivity to major and minor penicillin determinants and an increase in sensitivity to amoxicillin and others; this might reflect that the changes in prescription could have influenced the sensitization patterns, thus increasing the incidence of specific reactions to side-chain selective reactions.

An ultra-sensitive homologous chemiluminescence immunoassay to tackle penicillin allergy

Penicillin is one of the most widely used antibiotics to treat bacterial infections in clinical practice. The antibiotic undergoes degradation under physiological conditions to produce reactive compounds that in vivo bind self-proteins. These conjugates might elicit an immune response and trigger allergic reactions challenging to diagnose due to the complex immunogenicity. Penicillin allergy delabeling initiatives are now part of antibiotic stewardship programs and include the use of invasive and risky in vivo tests. Instead, the in vitro quantification of specific IgE is highly useful to confirm immediate allergy to penicillins. However, discrepant results associated with the low sensitivity and accuracy of penicillin allergy in vitro tests have limited their routine diagnostic use for delabeling purposes. We aimed to develop a homologous chemiluminescence-based immunochemical method for the reliable determination of specific IgE to penicillin G, using unprecedented synthetic human-like standards. The synthetic standard targets the major antigenic determinant of penicillin G and the paratope of Omalizumab, acting as human-like specific IgE. It is a potent calibrator, highly stable, easy, and inexpensive to produce, overcoming the limitations of the pooled human serum preparations. The developed method achieved a good agreement and strong positive relationship, reaching a detection limit below 0.1 IU mL−1 and excellent reproducibility (RSD <9%). The clinical sensitivity of the assay significantly increased (66%), doubling the accuracy of the reference method with an overall specificity of 100%. The new diagnostic strategy compares favorably with results obtained by the standard procedure, paving the way towards the standardization of penicillin allergy testing, and enhancing the detection sensitivity of specific IgE in serum to tackle reliably β-lactam allergy delabeling.

Studying the Adsorption of Penicillin (G) Antibiotic on the Surface of Carbon Nanotubes by Molecular Docking

Background: This research intended to evaluate the adsorption of single-walled carbon nanotubes (SCNT) with penicillin (G) antibiotic agent at the B3LYP/6-31G (d) theoretical level. Methods: First, we optimized penicillin (G) antibiotic and SCNT structures using the Gaussian program. Next, ten complexes with negative binding energy and steady-state than other cases were calculated using the molecular docking method and its ranking algorithm. Then, graphene was used to perform molecular orbitals calculations for the most stable configurations. Finally, binding energy and thermodynamic parameters were calculated at 298 K and 1 atm. Results: According to the findings, the adsorption of penicillin (G) antibiotic on the surface of carbon nanotubes was an exothermic, spontaneous process. Conclusions: This study indicated the ability of SCNT, as a sensing material, to construct thermal and electrochemical sensors for penicillin (G) determination.

Label-free optical sensor based on liquid crystal sessile droplet array for penicillin G determination

A sessile droplet array of 4’-pentyl-biphenyl-4-carboxylic acid (PBA) doped 5CB sessile droplets (12 −104 µm) is prepared by inkjet printing and applied for label-free sensing of penicillin G according to the configuration transition from axial to windmill based on the molecular alignment of 5CB at the LC-aqueous interface. Selective determination of penicillin G is achieved according to the target-selective enzymatic hydrolysis catalyzed by penicillinase, exhibiting sensitive detection and fast response with a detection limit of 0.178 ng/mL within 3–5 min. The sensitivity is linear depending on LC droplet size. Under the optimized conditions, the linear detection range is 1.78–178.0 ng/mL. Such a droplet array sensor has also been successfully applied to determine penicillin G in real river and lake water samples with RSD of 102–119%. Such a platform possesses the advantages of simple and cheap construction of an ordered array for multiplex sensing, and easy characterization via a polarizer. The quantification range of this proposed label-free optical penicillin G sensor covers the maximum residue limits in raw milk and river water defined in the European Union Regulation, and thus being applicable for food and environmental monitoring. By engineering LC-based probes, this approach could be extended to sense various molecules and ions in biomedical and chemical scenarios. And the sessile droplets on polymer substrates would also bring the possibility for employing in flexible and wearable devices in the future.

Synthesis of a Novel [diresorcinate‐1,10‐phenanthrolinecobalt(II)] Complex, and Potentiodynamic Fabrication of Poly(DHRPCo)/GCE for Selective Square Wave Voltammetric Determination of Procaine Penicillin G in Pharmaceutical and Biological Fluid Samples

AbstractThis work presents synthesis, and characterization of a novel [diresorcinate‐1,10‐phenanthrolinecobalt(II)] (DHRPCo) complex, and potentiodynamic fabrication of poly(DHRPCo)/GCE for SWV determination of procaine penicillin G (ProPenG) in various pharmaceutical and biological samples. While UV‐Vis and FT‐IR results demonstrated stepwise synthesis of DHRPCo, the CV and EIS results evidenced modification of the surface of GCE by an electroactive polymer film that improved electrode surface area. In contrast to the peak of ProPenG at the unmodified electrode, an irreversible oxidative peak at poly(DHRPCo)/GCE with lower potential and six folds of current enhancement observed, that confirmed electrocatalytic effect of the polymer, SWV response of poly(DHRPCo)/GCE showed linear dependence on concentration of ProPenG in the range 0.1–200 μM, with LoD of 4.9 nM. Spike recovery results in various pharmaceutical and biological samples in the range 98.03–106.50 %, interference recovery in the presence of selected potential interferents at their various levels with associated errors under 3.39 %, and stability of the modifier with analysis time validated the method for determination of ProPenG. Poly(DHRPCo)/GCE was applied for determination of ProPenG in three brands of injection pharmaceutical formulation, human urine, human blood serum, and cow's milk samples. While no ProPenG was detected in the human urine, serum, and cow's milk samples, detected ProPenG in the injection powder samples was within 96.1–101.3 % of company label. The developed method showed superior performance over recently reported methods making it an excellent candidate for determination of ProPenG in real samples.