Determination Of Drug Concentrations Research Articles

Pharmacokinetics of pradofloxacin, florfenicol, and tulathromycin and response to treatment of steers experimentally infected with Mannheimia hemolytica.

Bovine respiratory disease (BRD) is an economically important disease in the beef industry, and a major driver of therapeutic antibiotic use. Pharmacokinetic data of these drugs is relatively limited in diseased animals. To determine the concentrations of pradofloxacin, florfenicol, and tulathromycin in the airways, plasma, and interstitial fluid (ISF) of steers with a clinically relevant model of bacterial respiratory disease. Twenty-four Holstein and Holstein/Jersey cross steers ranging in age from 6 to 15 months. A randomized, blinded clinical trial was performed. After transport stress, steers were inoculated with Mannheimia hemolytica to induce BRD. Upon onset of clinical disease, steers were treated with pradofloxacin, florfenicol or tulathromycin. Blood, ISF, and pulmonary epithelial lining fluid (PELF) samples were obtained for drug concentration determination. Clinical exams and thoracic ultrasound examinations were conducted daily. Animals were euthanized at the end of the study period to assess lung lesions. Pradofloxacin Cmax in PELF was 0.81 μg/mL (CV = 49.02%) and penetration into the PELF was 203.58% (72%). Florfenicol Cmax in PELF was 2.94 μg/mL (42.1%) and penetration was 230.08% (78.82%). Tulathromycin PELF Cmax was 0.9 μg/mL (45.03%) and PELF penetration was 518.97% (56.59%). There are differences in penetration of the drugs into the ISF and PELF compared to one another and previous data from healthy steers demonstrating the effect of disease on the PK of these drugs.

Comparison of Different Approaches for Calculating LOD and LOQ in an HPLC-Based Analysis Method

Background: Sensitivity in the determination of the drug concentration is critical in pharmaceutical analysis. This research investigates several approaches for determining two sensitivity parameters, the Limit of Detection (LOD) and the Limit of Quantification (LOQ), in the analysis of the drug concentration using High-Performance Liquid Chromatography (HPLC). Methods: The study evaluates the FDA’s Lower Limit of Quantification (LLOQ) parameter, following global standards and quantitatively comparing sensitivity parameters for an established HPLC-UV method for the analysis of carbamazepine and phenytoin. Results: The study found that the LOD and LOQ values obtained by different methods varied significantly. The signal-to-noise ratio (S/N) method provided the lowest LOD and LOQ values for both drugs, while the standard deviation of the response and slope (SDR) method resulted in the highest values. This highlights the variability in sensitivity depending on the method used. Conclusion: The results show significant differences among calculated sensitivity values, emphasizing the influence of methodological variations on sensitivity values. It recommends following FDA criteria in chromatographic-based pharmaceutical analysis to improve the accuracy of drug concentration determination.

Developing Self-Nanoemulsifying Drug Delivery Systems Comprising an Artemether-Lumefantrine Fixed-Dose Combination to Treat Malaria.

Despite attempts to control malaria, poor drug bioavailability means malaria still places enormous pressure on health globally. It has been found that the solubility of highly lipophilic compounds can be enhanced through lipid formulations, e.g., self-emulsifying drug delivery systems (SEDDSs). Thus, quality-by-design and characterization were used to justify the development and determine the feasibility of oral oil-in-water SEDDSs comprising a fixed-dose combination (FDC) of artemether-lumefantrine to treat malaria more effectively without the aid of a fatty meal. These formulations were compared to a commercial product containing the same active compounds. Excipient compatibility and spontaneous emulsification capacity of different FDC-excipient combinations were identified by employing isothermal microcalorimetry, solubility, and water titration tests. Pseudoternary phase diagrams were constructed, and checkpoint formulations were selected within the self-emulsification region by reviewing formulation properties essential for optimized drug delivery. SEDDSs capable of enduring phase separation within 24 h were subjected to characterization experiments, i.e., drug concentration determination, cloud point, droplet size, size distribution, self-emulsification time, self-emulsification efficacy, viscosity, zeta potential, and thermodynamic stability analysis. SEDDSs with favorable characteristics were identified in the micro or nano range (SNEDDSs) before being subjected to drug release studies. All final formulations depicted enhanced artemether and lumefantrine release compared to the commercial product, which could not release lumefantrine at a quantifiable concentration in this study. The avocado oil (AVO)4:6 and olive oil (OLV)3:7 SNEDDSs overall portrayed the ideal characteristics and depicted the highest percentage of drug release. This study offers evidence that SNEDDSs from selected natural oils comprising an artemether-lumefantrine FDC can potentially enhance the bioavailability of these lipophilic drugs.

Development of SIMPATEC-USP: An abbreviated impactor for rapid testing of inhalation devices

Cascade impactors like the Andersen Cascade Impactor (ACI) and Next Generation Impactor (NGI) are complex and costly due to multiple stages. This study introduces SIMPATEC-USP (Simplified Impactor Developed at the University of São Paulo), a low-cost, single-stage alternative meeting US Pharmacopeia standards for testing dry powder inhalers (DPIs). SIMPATEC-USP simplifies particle retention into a single Petri dish stage, eliminating multi-stage complexity. Its design includes a straightforward closure system for easy assembly/disassembly. A collection chamber holds a glass Petri dish with filter paper for final filtration, ensuring efficient aerosol product collection. SIMPATEC-USP also offers potential use with culture media for applications like antibacterial screening. Operating at 30 L/min, SIMPATEC-USP consists of three parts: a single-stage chamber, an L-shaped tube mimicking the trachea, and a vacuum pump. Aerosol particles are deposited onto the Petri dish via a nozzle, and the collected sample is weighed to determine drug concentration. Tested with inhalation-grade lactose, SIMPATEC-USP effectively collects and analyzes particles, allowing for rapid aerodynamic comparison of formulations, capsule retention assessment, and sample collection for drug release studies. The results demonstrate that it is possible to evaluate the performance of three inhalation devices regarding the mass migrating to the collection plate and that retained within the device itself. In conclusion, SIMPATEC-USP is highly suitable for exploratory studies and educational activities in pharmaceutical technology and pulmonary drug delivery systems.

Ocular Surface Fluid: More than a Matrix.

Although the eye can be subjected to therapeutic manipulation, some of its structures are highly inaccessible. Thus, conventional therapeutic administration pathways, such as topical or systemic routes, usually show significant limitations in the form of low ocular penetration or the appearance of side effects linked to physiology, among others. The critical feature of many xenobiotics is the drug gradient from the concentrated tear reservoir to the relatively barren corneal and conjunctival epithelia, which forces a passive route of absorption. The same is true in the opposite direction, towards the ocular surface (OS). With the premise that tears can be regarded as equivalent to or a substitute for plasma, researchers may determine drug concentrations in the OS fluid. Within this framework, a survey of scholarly sources on the topic was conducted. It provided an overview of current knowledge, allowing the identification of relevant theories, methods, and gaps in the existing research that can be employed in subsequent research. OS fluid (tears particularly) has enormous potential as a source of biological material for external drug screening and as a biomarker of various systemic diseases. Given the numerous alternate matrices, knowledge of their properties is very important in selecting the most appropriate specimens in toxicological analyses.

Role of Chalcogenides in Sensitive Therapeutic Drug Monitoring Using Laser Desorption and Ionization.

This study investigates the applicability of six transition metal dichalcogenides to efficient therapeutic drug monitoring of ten antiepileptic drugs using laser desorption/ionization-mass spectrometry. We found that molybdenum ditelluride and tungsten ditelluride are suitable for the sensitive quantification of therapeutic drugs. The contribution of tellurium to the enhanced efficiency of laser desorption ionization was validated through theoretical calculations utilizing an integrated model that incorporates transition-metal dichalcogenides and antiepileptic drugs. The results of our theoretical calculations suggest that the relatively low surface electron density for the tellurium-containing transition metal dichalcogenides induces stronger Coulombic interactions, which results in enhanced laser desorption and ionization efficiency. To demonstrate applicability, up to 120 patient samples were analyzed to determine drug concentrations, and the results were compared with those of immunoassay and liquid chromatography-tandem mass spectrometry. Agreements among these methods were statistically evaluated using the Passing-Bablok regression and Bland-Altman analysis. Furthermore, our method has been shown to be applicable to the simultaneous detection and multiplexed quantification of antiepileptic drugs.

A Genome-Wide Association Study of Oxypurinol Concentrations in Patients Treated with Allopurinol.

Cohort studies have identified several genetic determinants that could predict the clinical response to allopurinol. However, they have not been commonly used for genome-wide investigations to identify genetic determinants on allopurinol metabolism and concentrations. We conducted a genome-wide association study of a prior cross-sectional investigation of patients from the Montreal Heart Institute Biobank undergoing allopurinol therapy. Four endpoints were investigated, namely plasma concentrations of oxypurinol, the active metabolite of allopurinol, allopurinol, and allopurinol-riboside, as well as allopurinol daily dosing. A total of 439 participants (mean age 69.4 years; 86.4% male) taking allopurinol (mean daily dose 194.5 mg) and who had quantifiable oxypurinol concentrations were included in the genome-wide analyses. Participants presented with multiple comorbidities and received concomitant cardiovascular medications. No association achieved the predefined genome-wide threshold values for any of the endpoints (all p > 5 × 10-8). Our results are consistent with prior findings regarding the difficulty in identifying genetic determinants of drug concentrations or pharmacokinetics of allopurinol and its metabolites, as well as allopurinol daily dosing. Given the size of this genome-wide study, collaborative investigations involving larger and diverse cohorts may be required to further identify pharmacogenomic determinants of allopurinol and measure their clinical relevance to personalize allopurinol therapy.

A developed and validated centrifugal ultrafiltration coupled with high performance liquid chromatography-tandem mass spectrometry method for rapid quantification of unbound lenvatinib in human plasma

In clinical practice, the determination of unbound drug concentration is very important for dose adjustment and toxicity prediction because only the unbound fraction can achieve a pharmacological effect. A fast, sensitive and accurate analytical method of centrifugal ultrafiltration coupled with high performance liquid chromatography-tandem mass spectrometry method was developed and applied to allow the quantification of unbound lenvatinib concentration. The application of linear regression analysis was used to examine the effects of centrifugal force, centrifugal time, and protein content on ultrafiltrate volume (Vu). The results indicated that the centrifugal force and centrifugal time have an influence on Vu that is significantly positive (P < 0.05). This developed method with good linearity (r2 = 0.9996), good accuracy (bias % ≤ 2.24 %), good precision (CV % ≤ 7.10 %), and good recovery (95.46 %−106.46 %) was suitable for routine clinical practice and studies. Particularly, the ultrafiltration membrane had no non-specific binding to lenvatinib. The unbound fractions can be separated in just 15 min. This method was applied to quantify clinical samples and to determine the plasma protein binding and unbound fraction of lenvatinib. This study provides a more effective and promising method for determination of unbound lenvatinib. It could be beneficial to measure the unbound concentration of lenvatinib in personalized medicine and therapeutic drug monitoring in routine clinical practice.

Association of CYP3A4*1B, CYP3A4*22 and CYP3A5*3 polymorphisms carriage with efficacy and safety of tamsulosin in patients with benign prostatic hyperplasia

Tamsulosin is a first-line drug in the treatment of lower urinary tract symptoms (LUTS) in benign prostatic hyperplasia (BPH). Despite high estimates of its efficacy and safety, it rates may vary due to genetic polymorphisms of genes for the enzymes involved in the drugs metabolism.The aim of the work was to evaluate the carriage influence of genes polymorphisms of the CYP3A enzymes group of tamsulosin metabolizers on the efficacy and safety of therapy in patients with LUTS in BPH.Materials and methods. A total of 142 patients with LUTS, with an established BPH diagnosis (N40 according to ICD-10) were included in the study and underwent all stages. All patients received monotherapy with tamsulosin 0.4 mg/day for at least 8 weeks. An IPSS questionnaire with the definition of quality of life, a prostate ultrasound with the determination of the prostate volume and residual urine, as well as uroflowmetry, were used to evaluate the results of the treatment. Controls were performed at 2, 4 and 8 weeks from the start of the therapy. The carriage of polymorphic markers CYP3A4 (*1B, *22) and CYP3A5*3 was determined in patients; HPLC was used to determine drug concentrations in blood plasma and levels of cortisol and its metabolite 6-beta-hydroxycortisol in urine to assess the phenotypic activity of CYP3A.Results. No statistically significant associations between CYP3A phenotype (defined by CYP3A4 and CYP3A5 genotypes) and clinical parameters of the tamsulosin therapy efficacy and the safety assessment in the studied sample of patients were found (p &gt;0.05). Similar data were obtained for individual variants of CYP3A4*1B, CYP3A4*22, CYP3A5*3 (p &gt;0.05). The comparison of the tamsulosin residual equilibrium concentration values in patients in the study sample with respect to the carriers of CYP3A4 and CYP3A5 gene variants did not reveal the presence of significant differences in either CYP3A phenotypes and carriers and non-carriers of individual CYP3A4*1B (p=0.57), CYP3A4*22 (p=0.37) and CYP3A5*3 (p=0.76) variants. No association was found between the metabolic ratio of 6-beta-hydroxycortisol / cortisol in urine and the CYP3A phenotype encoded by a combination of genotypes of CYP3A4 and CYP3A5 gene variants (p &gt;0.05).Conclusion. A possible association between the carriage of CYP3A4*1B, CYP3A4*22, CYP3A5*3 variants, a CYP3A activity assessed by the content of an endogenous substrate of this isoenzyme and its metabolite in urine, the level of plasma concentration of the drug, and the efficacy and safety of tamsulosin, has not been confirmed. The contribution of CYP3A4 and CYP3A5 genetic polymorphisms to clinical parameters of the tamsulosin therapy requires a further study.

Human β-defensin-1 affects the mammalian target of rapamycin pathway and autophagy in colon cancer cells through long non-coding RNA TCONS_00014506.

Colorectal cancer has a low 5-year survival rate and high mortality. Human β-defensin-1 (hBD-1) may play an integral function in the innate immune system, contributing to the recognition and destruction of cancer cells. Long non-coding RNAs (lncRNAs) are involved in the process of cell differentiation and growth. To investigate the effect of hBD-1 on the mammalian target of rapamycin (mTOR) pathway and autophagy in human colon cancer SW620 cells. CCK8 assay was utilized for the detection of cell proliferation and determination of the optimal drug concentration. Colony formation assay was employed to assess the effect of hBD-1 on SW620 cell proliferation. Bioinformatics was used to screen potentially biologically significant lncRNAs related to the mTOR pathway. Additionally, p-mTOR (Ser2448), Beclin1, and LC3II/I expression levels in SW620 cells were assessed through Western blot analysis. hBD-1 inhibited the proliferative ability of SW620 cells, as evidenced by the reduction in the colony formation capacity of SW620 cells upon exposure to hBD-1. hBD-1 decreased the expression of p-mTOR (Ser2448) protein and increased the expression of Beclin1 and LC3II/I protein. Furthermore, bioinformatics analysis identified seven lncRNAs (2 upregulated and 5 downregulated) related to the mTOR pathway. The lncRNA TCONS_00014506 was ultimately selected. Following the inhibition of the lncRNA TCONS_00014506, exposure to hBD-1 inhibited p-mTOR (Ser2448) and promoted Beclin1 and LC3II/I protein expression. hBD-1 inhibits the mTOR pathway and promotes autophagy by upregulating the expression of the lncRNA TCONS_00014506 in SW620 cells.

Simulated drug disposition in critically ill patients to evaluate effective PK/PD targets for combating Pseudomonas aeruginosa resistance to meropenem.

Bacterial infections, including those caused by Pseudomonas aeruginosa, often lead to sepsis, necessitating effective antibiotic treatment like carbapenems. The key pharmacokinetic/pharmacodynamic (PK/PD) index correlated to carbapenem efficacy is the fraction time of unbound plasma concentration above the minimum inhibitory concentration (MIC) of the pathogen (%fT > MIC). While multiple targets exist, determining the most effective one for critically ill patients remains a matter of debate. This study evaluated meropenem's bactericidal potency and its ability to combat drug resistance in Pseudomonas aeruginosa under three representative PK/PD targets: 40% fT > MIC, 100% fT > MIC, and 100% fT > 4× MIC. The hollow fiber infection model (HFIM) was constructed, validated, and subsequently inoculated with a substantial Pseudomonas aeruginosa load (1 × 108 CFU/mL). Different meropenem regimens were administered to achieve the specified PK/PD targets. At specified intervals, samples were collected from the HFIM system and subjected to centrifugation. The resulting supernatant was utilized to determine drug concentrations, while the precipitates were used to track changes in both total and drug-resistant bacterial populations over time by the spread plate method. The HFIM accurately reproduced meropenem's pharmacokinetics in critically ill patients. All three PK/PD target groups exhibited a rapid bactericidal response within 6 h of the initial treatment. However, the 40% fT > MIC and 100% fT > MIC groups subsequently showed bacterial resurgence and resistance, whereas the 100% fT > 4× MIC group displayed sustained bactericidal activity with no evidence of drug resistance. The HFIM system revealed that maintaining 100% fT > 4× MIC offers a desirable microbiological response for critically ill patients, demonstrating strong bactericidal capacity and effective prevention of drug resistance.

Hollow-fiber liquid phase microextraction for determination of fluoxetine in human serum by nano-liquid chromatography coupled to high resolution mass spectrometry

Therapeutic drug monitoring (TDM) is a personalized care tool based on the determination of a target drug concentration in human serum. An antidepressant drug of interest for such investigations is fluoxetine (FXT), due to a severe impact of genetic polymorphisms on its metabolism. A bioanalytical method employed for TDM purposes must exhibit satisfactory selectivity and detectability, which becomes more difficult due to highly complex biological matrices. In this study, a highly selective bioanalytical method for the determination of FXT in human serum is proposed, which provides excellent clean-up efficiency based on a low cost hollow fiber liquid-phase microextraction (HF-LPME) sample preparation step and nano-liquid chromatography coupled to high-resolution mass spectrometry (nano-LC-HRMS). HF-LPME was performed using a two-phase “U” configuration, with 6 cm fiber, 20 µL of 1-octanol acting as supported liquid membrane, and ammonium hydroxide (pH 10) as the donor phase with NaCl (10 % m/v) and methanol (5 % v/v) as additives, requiring only 250 µL of the sample. The procedure was conducted for 30 min under a 750 rpm stirring rate. Gradient elution was carried out employing an acetonitrile–water as mobile phase, the composition of 30:70 to 100:00 (v/v) for 15 min, using formic acid 0.1 % (v/v) as an additive. MS1 was acquired in an Orbitrap mass analyzer, while MS2 was acquired in a linear trap quadrupole. Satisfactory linearity (Pearson’s r = 0.99709) was obtained for a concentration range of 0.02 to 2.5 µg mL−1, which is compatible with the therapeutic and toxic range for FXT. The developed method presents adequate precision (1.61 to 7.45 %) and accuracy (95 to 114 %) and allows the dilution of high concentration samples in a 1:4 ratio (v/v), enabling its application for forensic serum samples. To our knowledge, this is the first study reporting a method based on HF-LPME and nano-LC-HRMS with any analytical purpose, especially with a TDM focus.

DNA hybrid florescent chrominance sensor: Drug sensing through direct gradient florescent color transformation of hybrid DNA crystals loaded lanthanide (Eu3+/Tb3+) complexes

In this study, a highly sensitive anticancer drug sensor was synthesized using DNA crystals loaded with luminescent Ln3+(Eu3+/Tb3+) complexes. The sensors exhibited a pronounced color transformation property due to the differential quenching of Eu3+ and Tb3+ emissions upon drug interactions. These luminescent hybrid DNA crystals present a significant advancement in sensing anticancer drugs like curcumin, transitioning from monitoring fluorescence intensity changes to a visually discernable color change. This new approach eliminates the need for specialized instrumentation. Owing to the intrinsic biocompatibility of materials composed of DNA, this hybrid sensor has the potential to sense drugs within the human body. Hybrid crystals loaded with a single complex will lose their fluorescence properties due to drug quenching; however, crystals loaded with mixed complexes emit distinct fluorescent colors, such as yellow-green and orange-red, at different drug concentrations. This color transformation allows for the direct determination of drug concentration based on crystal color, making the sensing process more simple and easily observable.

Salt-assisted liquid–liquid microextraction for determination of haloperidol in human plasma by LC-MS/MS

Haloperidol is an antipsychotic used in the treatment of schizophrenia. Compared to other antipsychotics, it is widely used in developing countries due to its affordable price. Haloperidol has a narrow therapeutic range and variable pharmacokinetics; therefore, therapeutic drug monitoring (TDM) is recommended. For this reason, in this study, an easily applicable, fast, selective, accurate, reliable, and economical LC-MS/MS method was developed for the determination of haloperidol in human plasma for use in TDM and also method was validated according to European Medicines Agency (EMA) Bioanalytical method validation guidelines. In the developed method, analyte and internal standard were extracted from plasma by salt-assisted liquid-liquid microextraction (SALLME) technique and after that injected to the LC system. The limit of quantification of haloperidol was determined as 1 ng/ml. The calibration curve was validated between 1-15 ng/ml, with correlation coefficients &gt;0.99. In addition, the developed method was used to determine drug concentration levels in the plasma of real patients.

Label-free mapping of cetuximab in multi-layered tumor oral mucosa models by atomic force-microscopy-based infrared spectroscopy.

Sensitive mapping of drugs and drug delivery systems is pivotal for the understanding and improvement of treatment options. Since labeling alters the physicochemical and potentially the pharmacological properties of the molecule of interest, its label-free detection by photothermal expansion is investigated. We report on a proof-of-concept study to map the cetuximab distribution by atomic-force microscopy-based infrared spectroscopy (AFM-IR). The monoclonal antibody cetuximab was applied to a human tumor oral mucosa model, consisting of a tumor epithelium on a lamina propria equivalent. Hyperspectral imaging in the wavenumber regime between 903 cm-1 and 1312 cm-1 and a probing distance between the data points down to 10 × 10 nm are used for determining the local drug distribution. The local distinction of cetuximab from the tissue background is gained by linear combination modeling making use of reference spectra of the drug and untreated models. The results from this approach are compared to principal component analyses, yielding comparable results. Even single molecule detection appears feasible. The results indicate that cetuximab penetrates the cytosol of tumor cells but does not bind to structures in the cell membrane. In conclusion, AFM-IR mapping of cetuximab proved to sensitively determine drug concentrations at an unprecedented spatial resolution without the need for drug labeling.

Comparative Analysis of Sibutramine Concentration Determination

This paper underscores the practical implications of employing FTIR RMSE analysis in pharmaceutical research. It highlights the method's rapid and non-destructive nature for quantifying drug concentration in complex formulations, emphasizing its significance in pharmaceutical quality assurance. The study advocates for integrating FTIR RMSE with conventional gravimetric and volumetric methods to enhance the analytical toolkit for pharmaceutical compound analysis. Furthermore, it elucidates the potential of FTIR RMSE in expediting analytical processes and improving estimation accuracy. The paper accentuates the synergy between classical gravimetric techniques and modern spectroscopic methods, providing valuable insights into pharmaceutical compound analysis within heterogeneous formulations. In terms of contributions, this paper advances analytical methods in pharmaceutical research by comprehensively exploring traditional gravimetric/volumetric techniques alongside Fourier Transform Infrared Spectroscopy (FTIR) coupled with Root Mean Square Error (RMSE) analysis. It assesses their effectiveness in determining drug concentrations, mainly focusing on sibutramine quantification, a complex active pharmaceutical ingredient. The results indicate a strong correlation (R² = 0.987) between the two methodologies, with FTIR RMSE consistently yielding slightly higher results due to its sensitivity to molecular structural fluctuations and component interactions. In conclusion, this study advocates for integrating FTIR RMSE into pharmaceutical research practices. It emphasizes the need for calibration and refinement of this technique to further enhance estimation accuracy and analytical precision. It underscores the pivotal role of advanced analytical methods in various applications, including pharmaceutical and biomedical fields.

Developing an in vitro lipolysis model for real-time analysis of drug concentrations during digestion of lipid-based formulations

Understanding the effect of digestion on oral lipid-based drug formulations is a critical step in assessing the impact of the digestive process in the intestine on intraluminal drug concentrations. The classical pH-stat in vitro lipolysis technique has traditionally been applied, however, there is a need to explore the establishment of higher throughput small-scale methods. This study explores the use of alternative lipases with the aim of selecting digestion conditions that permit in-line UV detection for the determination of real-time drug concentrations. A range of immobilised and pre-dissolved lipases were assessed for digestion of lipid-based formulations and compared to digestion with the classical source of lipase, porcine pancreatin. Palatase® 20000 L, a purified liquid lipase, displayed comparable digestion kinetics to porcine pancreatin and drug concentration determined during digestion of a fenofibrate lipid-based formulation were similar between methods. In-line UV analysis using the MicroDISS ProfilerTM demonstrated that drug concentration could be monitored during one hour of dispersion and three hours of digestion for both a medium- and long-chain lipid-based formulations with corresponding results to that obtained from the classical lipolysis method. This method offers opportunities exploring the real-time dynamic drug concentration during dispersion and digestion of lipid-based formulations in a small-scale setup avoiding artifacts as a result of extensive sample preparation.

Electrochemical sensor based on polypyrrole/triiron tetraoxide (PPY/Fe3O4) nanocomposite deposited from a deep eutectic solvent for voltammetric determination of procaine hydrochloride in pharmaceutical formulations

This investigation reports the electrochemical preparation of a polypyrrole/triiron tetraoxide (PPY/Fe3O4) nanocomposite from deep eutectic solvents (DESs), and their application as novel sensor electrodes to determine certain pharmaceutical formulations using cyclic voltammetry (CV). The modified polypyrrole/Fe3O4 nanocomposite was characterized using transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDAX), X-ray diffraction (XRD), and Fourier-transform infrared (FTIR) spectroscopy. These techniques confirmed the incorporation of Fe3O4 into the polypyrrole composite. Cyclic voltammetry and electrochemical impedance spectroscopy (EIS) indicated that PPY/Fe3O4 shows higher redox currents and greater electrical conductivity with procaine hydrochloride than other pharmaceutical formulations (ascorbic acid, diclofenac, losartan potassium, adiphenine hydrochloride, and metformin hydrochloride). Factors that could influence the determination of procaine hydrochloride, including pH, temperature, scan rate, concentration of drug, stability, and reproducibility, have all been optimized. Under optimal conditions, the calibration curve shows a linear relationship between oxidation peak current and procaine hydrochloride concentrations (between 0.005 and 0.3 mol/L), with an R2 of 0.9943. The limit of detection (LOD) was 0.0024 mol/L and the limit of quantification (LOQ) was 0.0079 mol/L. The modified sensor (PPY/Fe3O4) constructed in this study was successfully used for the determination of procaine hydrochloride drug concentrations in a procaine-penicillin pharmaceutical product.

Serum levels of infliximab and adalimumab are associated with deep remission in inflammatory bowel disease

ObjectiveDeep remission defined by clinical-biomarker remission and mucosal healing has emerged as a new therapeutic target in inflammatory bowel disease. The aim of this study was to define an optimal cut-off concentration for infliximab and adalimumab during maintenance therapy associated with deep remission. The secondary objective, was to evaluate the influence of variables on anti tumor necrosis factor-alpha concentrations and deep remission. MethodRetrospective study including 120 and 122 patients inflammatory bowel disease diagnosed who received maintenance therapy with infliximab and adalimumab. Biomarker remission was considered by C-reactive protein < 5 mg/L and fecal calprotectin < 100 µg/g. Crohn's disease clinical remission was defined by a Harvey Bradshaw score < 5 and mucosal healing by a simple endoscopic score for Crohn's disease< 3. In ulcerative colitis, it was defined as a Mayo total score < 3 and Mayo endoscopic subscore < 2. Receiver operating characteristic test was performed to determine drug concentration thresholds associated with deep remission. Anti tumor necrosis factor-alpha concentrations were classified into quartiles. X2 and Kruskal-Wallis test were used to compare discrete and continuous variables between quartile groups. Multivariate logistic regression was performed to identify patient characteristics and serological facto C-reactive protein rs associated with deep remission. ResultsAnti tumor necrosis factor-alpha concentrations were higher in patients with deep remission, in infliximab (4.4, interquartile range: 3.3-6.5 vs 2.3, interquartile range: 1.1-4.2 µg/mL, P < 0.005) and adalimumab (6.3, interquartile range: 4.2-8.2 vs 3.9, interquartile range: 2.4-5.5 µg/mL, P < 0.005). A Receiver operating characteristic test identified a concentration threshold of 3.1 µg/mL in infliximab (area under the Receiver operating characteristic test curve, 0.72) and 6.3 µg/mL in adalimumab (area under receiver operating characteristic test curve, 0.75) associated with deep remission. Factors associated with the highest quartiles of serum infliximab concentration were: elevated body mass index, absence of previous inflammatory bowel disease-surgery, C-reactive protein < 5 mg/L, and fecal calprotectin < 100 µg/g. In adalimumab, higher quartiles were related to concomitant immunosuppressants, low body mass index, absence of previous inflammatory bowel disease-surgery, and C-reactive protein < 5 mg/L and fecal calprotectin < 100 µg/g. Multivariate regression identified fecal calprotectin < 100 µg/g, C-reactive protein < 5 mg/L, infliximab > 3.1 µg/mL and adalimumab concentrations > 6.3 µg/mL as factors significantly associated with deep remission. ConclusionsTrough infliximab and adalimumab concentrations, C-reactive protein < 5 mg/L and fecal calprotectin < 100 µg/g are associated with deep remission during maintenance therapy. Cutoff point of 3.1 and 6.3 g/mL for infliximab and adalimumab respectively, were identified as deep remission predictors.

Protein binding investigation of first-line and second-line antituberculosis drugs

Data about antituberculosis drugs binding are incomplete for first-line drugs and lacking for second-line drugs that are used extensively for multi-drug resistant tuberculosis (levofloxacin, linezolid and moxifloxacin). Thus, the main purposes of this study were first to investigate thoroughly the relation between the carrier-proteins level and the drug binding and second to investigate the feasibility of predicting the free drug concentrations by the means of in vitro and in vivo results. In vitro experiments mimicked real-case samples by spiking drugs combinations from the clinical practice. We measured a median in vivo protein binding of 1.5% for ethambutol, 9.7% for isoniazid, 0.7% for pyrazinamide and 88.2% for rifampicin; and a median in vitro protein binding of 26.2% for levofloxacin, 12.8% for linezolid and 46.3% for moxifloxacin. The albumin concentration had a moderate impact on the moxifloxacin binding and a strong impact on the levofloxacin, linezolid and rifampicin binding (below values of 30 g/L). The determination of the free drug concentration seems to have few interests for ethambutol, isoniazid, moxifloxacin and pyrazinamide, a limited interest for linezolid because of its low binding and a major interest for rifampicin in hypoalbuminemia TB patients and for levofloxacin because their total concentration was an inaccurate reflection of the free concentration. The free concentration predicted by mathematical model was suitable for levofloxacin and linezolid, unlike for rifampicin, where the real free concentration should be measured. Further investigations should be carried out to investigate the benefit of the free concentration for levofloxacin, linezolid and rifampicin mainly in the critical period of active tuberculosis associated to hypoalbuminemia.