A versatile ratiometric fluorescence nanoprobe for the determination of clonazepam in patients' plasma samples.

Despite the necessity of the study of therapeutic drug monitoring of clonazepam (CLZ), there are only a few fast detection methods available for determining CLZ in biological media. This study aims to develop a cost-effective and ratiometric probe for the quantification of CLZ in plasma samples. Fluorescent polydopamine nanoparticles were produced through a self-polymerization process at a pH of 8.5. Rhodamine B molecules were employed as a fluorescent reference material, emitting stable fluorescence in the visible range. The fabricated probe exhibited a specific detection capability for CLZ. The fluorescence emission of the probe was enhanced in two concentration ranges: from 50 ng/mL to 1.0 µg/mL and from 1.0 to 15.0 µg/mL with a lower limit of quantification of 50 ng/mL, indicating the sensitivity of the probe for detecting CLZ plasma levels. The accuracy of the probe is favorable which could be recommended for CLZ monitoring in the biological media. Furthermore, this probe is highly specific towards CLZ in the presence of various interfering agents which is mainly caused by its ratiometric nature. The developed platform showed high reliability in quantifying CLZ concentrations in patients' plasma samples. Hence, the fabricated probe could be recommended as a reliable method for the routine detection of CLZ in clinical settings.

The influence of advanced materials on the analytical performance of semiconductor-based gas sensors.

Chemiresistive gas sensors are metal oxide-based sensors that have received significant attention in different fields. Ambient gas sensors are especially important in the fabrication of wearable probes for the real-time detection of biomarkers in human body samples. Usually, room temperature sensors are affordable due to their low power consumption, resulting in simple instrumentation and maintenance. To fabricate versatile gas sensors, i.e. sensitive, selective, ambient temperature operating gas sensors, and improve the sensing performance of the traditionally used sensor, new materials play an important role. In other words, new advanced materials are essential for designing and fabricating new gas sensors. Hence, in this review, the application and impact of new advanced materials in the fabrication of reliable gas sensors are discussed in detail. Special emphasis is given to the effect of new materials in the fabrication of room-temperature operating systems. Finally, future research outlook and possible challenges that may be encountered by reliable gas sensors are also explained.

Dispersive solid-phase extraction of risperidone from plasma samples using graphene oxide aerogels and determination with liquid chromatography.

A graphene oxide-based aerogel was synthesized and applied to the extraction and the determinations with the high-performance liquid chromatography-ultraviolet detector. After the characterization of the produced graphene-aerogel, it was utilized as a dispersive solid-phase extraction sorbent for risperidone extraction from plasma samples. Aerogels are materials with a large surface area-to-mass ratio and plenty of core with functional groups which can easily attach to the analytes to extract them to the second phase. The suggested method determined risperidone in plasma samples in the wide dynamic range from 20 ng/ml to 3 µg/ml. The limits of detection and quantification of the developed method were calculated as 2.4 and 8.2 ng/ml, respectively. As a novel feature, the developed method has no need to precipitate plasma proteins, improving the analytical performance of the analysis. Also, for the first time, the produced materials were utilized for the extraction of risperidone from the plasma samples. The obtained results revealed that the developed approach could be employed as an accurate method for the quantification of risperidone in real plasma samples.

Detection of pioglitazone based on dual-emission ratiometric fluorescence probe consisting of ZIF8 and to L-ASC-AuNP/DA nanoparticles.

A simple and sensitive dual-emission ratiometric fluorescent probe was developed using zeolitic imidazolate framework 8 (ZIF8) and L-ascorbic acid Au-doped dopamine nanoparticles (ZIF8/L-ASC-AuNP/DA NP) for the determination of pioglitazone (Pio), an oral hypoglycemic agent and insulin sensitizer, in real samples. The prepared system was based on the Pio-enhanced dual-emission intensity of ZIF8/L-ASC-AuNP/DA NP. The potential impact of various parameters on the system's emission intensity was tested. According to the findings, there is a strong linear correlation between the system's turn-on fluorescence intensity and Pio concentrations in the range 0.3 nM to 30.0 µM. The obtained value for the limit of detection (LOD) was 0.14 nM. In addition, the intra- and inter-day accuracy of the nanoprobe was studied and the findings revealed satisfactory precision and accuracy of the system. The short-term and freeze-thaw stability of Pio in plasma samples was evaluated and the results indicated the high stability of the developed nanoprobe under the test conditions. Pio was accurately detected in human plasma samples under ideal conditions with analytical recoveries in the range 86.0 - 109.3%. The results showed that the devised probe may be employed as an easy, sensitive, and precise approach for detecting Pio in real samples.

Two-photon fluorescence probe for quantification of cyclosporine.

A novel ratiometric fluorescent sensor was fabricated for the fast and facile determination of cyclosporine A (CsA). Due to the narrow therapeutic index of CsA, its desired therapeutic effects are evident within a limited range of blood concentration, indicating the fundamental role of therapeutic drug monitoring in CsA pharmacological response. In this study, a two-photon fluorescence probe based on the zeolitic imidazolate framework (ZIF-8) and norepinephrine-capped silver nanoparticles (AgNPs@NE) was employed for the quantification of the CsA in human plasma samples. In the presence of CsA, the fluorescent emission intensity of ZIF-8-AgNPs@NE was quenched. Under the optimum conditions, the proposed probe determines CsA in plasma samples in two linear ranges of 0.01 to 0.5 µg mL-1 and 0.5 to 10 µg mL-1. The developed probe demonstrates the advantages of a facile and fast platform with limit of detection as low as 0.007 µg mL-1. At last, this method was applied to find CsA concentration in four patients receiving oral CsA regimen which indicates it as a promising method for on-site detection applications.

Applications of scaffold-based advanced materials in biomedical sensing.

There have been many efforts to synthesize advanced materials that are capable of real-time specific recognition of a molecular target, and allow the quantification of a variety of biomolecules. Scaffold materials have a porous structure, with a high surface area and their intrinsic nanocavities can accommodate cells and macromolecules. The three-dimensional structure (3D) of scaffolds serves not only as a fibrous structure for cell adhesion and growth in tissue engineering, but can also provide the controlled release of drugs and other molecules for biomedical applications. There has been a limited number of reports on the use of scaffold materials in biomedical sensing applications. This review highlights the potential of scaffold materials in the improvement of sensing platforms and summarizes the progress in the application of novel scaffold-based materials as sensor, and discusses their advantages and limitations. Furthermore, the influence of the scaffold materials on the monitoring of infectious diseases such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and bacterial infections, was reviewed.

Synthesis of folic acid functionalized terbium-doped dendritic fibrous nano-silica and Interaction with HEK 293 normal, MDA breast cancer and HT 29 colon cancer cells.

A novel folic acid functionalized terbium-doped dendritic fibrous nanoparticle (Tb@KCC-1-NH2 -FA) with high surface area was synthesized using a novel hydrothermal protocol. In the present work, we report the fluorescent Tb-doted nanomaterial with emission wavelength at 497 nm which confirms the formation of Tb@KCC-1-NH2 -FA. Synthesized nanoparticles were investigated through transmission electron microscope, field emission scanning electron Microscopy, Fourier transform infrared spectra, Brunauer-Emmett-Teller, energy dispersive X-ray, Zeta potential and particle size distribution values and AFM (Atomic force microscopy) techniques. Specially, our desired nanomaterial which has FA moieties on the surface of Tb@KCC-1-NH2-FA where interact with folate receptor (FR) which there is on the surface of the various cancer cells. For this purpose, fluorescence microscopy images were used to prove the uptake of FA based nanomaterial with FR-positive MDA breast cancer and HT 29 colon cancer cells. Also HEK 293 normal cells as FR-negative cells verified the specificity of our desired nanomaterial toward the FR-positive cells. The cytotoxicity survey of Tb@KCC-1-NH2 -FA was examined by MTT assays against MDA breast cancer, HT 29 colon cancer and HEK 293 Normal cell lines which confirmed their biocompatible nature with any significant cytotoxic effects even for concentration higher than 900 µg/mL which could be used as a non-toxic catalyst or carrier in biological ambient. Hence, Tb@KCC-1-NH2 -FA were synthesized using green and hydrothermal method; the process was simple with good productivity and desired nanocomposite was non-toxic.

A dual-emission ratiometric fluorescent biosensor for ultrasensitive detection of glibenclamide using S-CDs/CdS quantum dots.

In the present work, sulfide-doped carbon dots (S-CDs)/cadmium sulfide quantum dots (CdS QDs) ratiometric fluorescent nanosensor has been developed for sensitive and selective determination of glibenclamide (GLC) in biological fluids. The method was based on the quenching effect of GLC on the dual-emission intensity of the S-CDs/CdS QDs system at 420 nm and 650 nm, which are related to S-CDs and CdS QDs, respectively. The fluorimetric data analysis indicated that the fluorescence signals of the system were quenched by adding GLC in a concentration-dependent manner. A good linear relationship was observed between GLC concentration and the quenched fluorescence intensity of the S-CDs/CdS QDs in the range of 0.3 nM-10.0 µM. The limit of detection (LOD) value was estimated to be 0.12 nM. Furthermore, under optimum conditions, GLC was detected in spiked human serum sample (as real media) using the developed ratiometric nanosensor with an accuracy of 99.6%. According to the results, the developed dual-emission system can be used as a reliable method for the quantitative detection of GLC in biological samples.

Applications of Advanced Materials for Non-Enzymatic Glucose Monitoring: From Invasive to the Wearable Device.

Diabetes mellitus (DM) is a global health problem leading to many complications and disabilities in life adjusting activities and even dead. Monitoring glucose levels is a key factor in diagnosis and management of DM. Conventional glucose sensors consisted of immobilized enzymes, are so susceptible to environmental conditions. In this way, nonenzymatic biosensors have attracted extensive attentions in many clinical diagnostics applications. To date, the finger pricking test is a common enzyme-based glucometer that is an invasive and inconvenient and may lead to infections in the injection sites. So, working on the possibility of cutaneous or subcutaneous insertion of devices as a noninvasive or minimally-invasive systems for continuous glucose controlling approaches through human biofluids (blood, perspiration, tears, saliva, etc.) have stimulated growing interest. This review summarizes recent nonenzymatic and noninvasive biofluids glucose monitoring systems which are highly resilience and stretchable to continuously adapt to body movements during common physical activity. Sensors are based on their constituent materials including carbon-based, metal nanoparticles, polymer, and hydrogel systems are classified for electrochemical, and optical glucose detection. Finally, we address the drawbacks and challenges of enzyme-free sensors which are aroused sustaining research passion to be used in point-of-care medical diagnostics applications.

Electrochemical sensing of doxorubicin hydrochloride under sodium alginate antifouling conditions using silver nanoparticles modified glassy carbon electrodes.

Doxorubicin (DOX) is a highly effective anticancer drug with a narrow therapeutic window; thus, sensitive and timely detection of DOX is crucial. Using electrodeposition of silver nanoparticles (AgNPs) and electropolymerization of alginate (Alg) layers on the surface of a glassy carbon electrode, a novel electrochemical probe was constructed (GCE). The fabricated AgNPs/poly-Alg-modified GCE probe was utilized for the quantification of DOX in unprocessed human plasma samples. For the electrodeposition of AgNPs and electropolymerization of alginate (Alg) layers on the surface of GCE, cyclic voltammetry (CV) was used in the potential ranges of -2.0 to 2.0 V and -0.6 to 0.2 V, respectively. The electrochemical activity of DOX exhibited two oxidation processes at the optimum pH value of 5.5 on the surface of the modified GCE. The DPV spectra of poly(Alg)/AgNPs modified GCE probe toward consecutive concentrations of DOX in plasma samples demonstrated wide dynamic ranges of 15 ng/mL-0.1 µg/mL and 0.1-5.0 µg/mL, with a low limit of quantification (LLOQ) of 15 ng/mL. The validation results indicated that the fabricated electrochemical probe might serve as a highly sensitive and selective assay for the quantification of DOX in patient samples. As an outstanding feature, the developed probe could detect DOX in unprocessed plasma samples and cell lysates without the requirement for pretreatment.

In-situ preparation of norepinephrine-functionalized silver nanoparticles and application for colorimetric detection of tacrolimus in plasma samples.

Tacrolimus (Tac) is a well-documented immunosuppressive agent for the prevention of graft-vs-host diseases in several types of organ transplants. The narrow therapeutic window and the individual-variable pharmacokinetics of Tac demonstrate the importance of regular therapeutic drug monitoring (TDM) as an imperative concept for its oral medication regimens. A simple, one-step, selective, and sensitive colorimetric platform is fabricated for the determination of Tac by surface modification of the silver nanoparticles (AgNPs) via norepinephrine (NE) molecules. The attachment of NE and Tac induces the aggregation of the AgNPs, which is observed by color distinction (yellow to brown) and a noteworthy shifting of the absorption peak in the visible region. The fabricated nanoprobe can detect Tac concentrations in plasma samples in two linear ranges from 2 ng/mL to 70 ng/mL and 70 ng/mL to 1000 ng/mL with R2 > 0.99. The limit of detection (LOD) was calculated as low as 0.1 ng/mL. The developed method was applied for the determination of Tac in patient's plasma samples under Tac medication therapy.

Synthesis and characterization of folate-functionalized silica-based materials and application for bioimaging of cancer cells.

Early-stage detection is a vital factor in the later treatment and prognosis of cancer. Enhancing the sensitivity and specificity of the cancer detection pathological and experimental approaches can affect the morbidity and mortality of this disease. A folic acid (FA)-functionalized silica quantum dots (SiQDs)/KCC-NH2@SiO2 nanomaterials were synthesized and characterized as a bioimaging agent of the MCF 7 cancer cells. These nanoparticles showed biocompatible nature with specificity towards folate receptor (FR)-overexpressed MCF 7 cancer cells. Viability findings suggested that the SiQDs/KCC-NH2@SiO2/FA nanomaterials have nontoxic nature towards the cells in the concentration of 200 µg/mL. Fluorescence microscopy images were utilized to estimate the cell internalization of the nanoparticles and further verified by the flow cytometry technique. The differentiation ability of the nanoparticles was also approved by incubation with FR-negative HEK 293 normal cells. The SiQDs/KCC-NH2@SiO2/FA nanoparticle exhibited high stability, bright and high quantum yield fluorescence emission, proposing as a high-quality material for in vivo bioimaging of FR-overexpressed circulating tumoral cancer cells (CTCs).

Serum Levels of Indoxyl Sulfate and P-cresol in Type II Diabetic Patients With and Without Nephropathy.

INTRODUCTION: Indoxyl sulfate (IS) and para-cresol (p-cresol) are uremic toxins with high protein bonding index that accumulate in the body with decreasing kidney function. The main purpose of the current investigation was to compare the concentration of p-cresol and IS in serum of the type II diabetic individuals with and without nephropathy. METHODS: Fifty-five patients with type II diabetes mellitus were divided into two groups: case and control. The case group consisted of 26 diabetic patients with nephropathy (proteinuria and serum creatinine below 1.5 mg/dL) without any other kidney diseases. The control group included 29 patients without diabetic nephropathy. Patients with advanced heart disease, cerebrovascular accident and other inflammatory or infectious diseases were excluded. Five mL of venous blood was taken from each patient in the morning fasting state. Then other laboratory tests including serum uric acid and creatinine levels, serum urea nitrogen, lipids and glucose were measured by standard methods. P-Cresol and IS levels were measured by the spectrofluorimetric method after extraction. We also filled out a checklist with information regarding the duration of their disease, medication history (oral or injectable), and other demographic information. There were no significant differences between the two groups regarding the investigated factors Results. There were no significant difference among the investigated factors between the two groups (P > .05) except for the serum creatinine, proteinuria and estimated glomerular filtration rate, where the mean values of cases were considerably higher than those of the controls. Serum IS and p-cresol levels were also significantly higher in the case group (P < .05). CONCLUSION: According to the findings, it seems that IS, and p-cresol may play a role in the development of diabetic nephropathy and other complications of diabetes mellitus.  DOI: 10.52547/ijkd.7266.

A ratiometric electrochemical probe for the quantification of apixaban in unprocessed plasma samples using carbon aerogel/BFO modified glassy carbon electrodes.

A novel electrochemical probe was established for the quantification of apixaban (APX) in unprocessed plasma samples. Efficiently oxidized graphene oxide aerogels (EEGO-AGs) and nano-sized Bi2Fe4O9 (BFO) particles were electrodeposited on the surface of a glassy carbon electrode (GCE). In this work, a ratiometric electrochemical method was introduced for APX detection to enhance the specificity of the probe in plasma samples. The fabricated ratiometric probe was employed for the indirect detection determination of APX using K3[Fe(CN)6]/K4[Fe(CN)6] as the redox pair. The differential pulse voltammetry technique was used to record the current alteration of the BFO/EEGO-AG-functionalized GCE probe at various APX concentrations. The probe response was proportional to the APX concentrations from 10 ng mL-1 to 10 µg mL-1 with a low limit of quantification (LLOQ) of 10 ng mL-1. After validation, this method was successfully utilized for the determination of APX in patients' plasma samples who have taken APX regularly. The fabricated chemosensor detected APX concentrations in unprocessed plasma samples with high selectivity, resulting from the physical filtering antifouling activity of aerogels.

Spectroscopic aspects on the interaction of nisin with serum albumin: thermodynamic and kinetic studies.

Introduction: Nisin is a bacteriocin produced by Streptococcus and Lactococcus species and has antimicrobial activity against other bacteria. Nisin omits the need to use chemical preservatives in food due to its biological preserving properties. Methods: In the present in vitro study, we investigated nisin interaction with bovine serum albumin (BSA) using fluorescence spectroscopy and surface plasmon resonance (SPR) analysis to obtain information about the mechanisms of BSA complex formation with nisin. Results: The BSA fluorescence intensity values gradually diminished with rising nisin concentration. The BSA fluorescence quenching analysis indicated that a combined quenching mechanism plays the main role. Finally, the Kb values were reduced with increasing temperature, which is demonstrative of nisin-BSA complex stability decrease at high temperatures. The negative values of ΔH° and ΔS° showed that hydrogen bonds and van der Waals forces are the foremost binding force between BSA and nisin. Meanwhile, the negative values of ΔG° demonstrated the exothermic and random nature of the reaction process. The results of the SPR verified the gained results through the fluorescence spectroscopy investigation, which denoted that the BSA affinity to nisin diminished upon increasing temperature. Conclusion: Overall, fluorescence spectroscopy and SPR results showed that the BSA interaction with nisin decreased with rising temperatures.

Determination of deferiprone in urine and serum using a terbium-sensitized luminescence method.

Optimized conditions, validation and practical applications of a new, rapid and specific fluorometric method for the determination of deferiprone (DFP) in urine and serum samples are reported. The proposed method, which is based on the formation of a luminescent complex with Tb(3+) ion, is evaluated in terms of linearity, accuracy, precision, stability, recovery and limits of detection (LOD) and quantification (LOQ). Under optimum conditions (pH 7.5, [Tb(3+)] = 3 × 10(-4) mol/L, temperature 0 °C and excitation wavelength 295 nm), the relative intensities at 545 nm are linear, with the concentration of DFP in the range 0.072-13 mmol/L for urine and serum samples. The LOD and LOQ, respectively, are calculated to be 0.014 and 0.045 mmol/L for urine and 0.022 and 0.072 mmol/L for serum samples. The intra-day and inter-day values for the precision and accuracy of the proposed method are all < 5%, and the recovery of the method is in the range 97.1-103.8%. The method was applied to human urine and serum samples collected from patients receiving DFP. The results indicated that the method can be successfully applied to the determination of DFP in human urine and serum samples collected for clinical or biopharmaceutical investigations in which simple, rapid, cheap and specific determination methods facilitate and speed up the analytical procedure.

Rational design of smart nano-platforms based on antifouling-nanomaterials toward multifunctional bioanalysis.

The ability to design nanoprobe devices with the capability of quantitative/qualitative operation in complex media will probably underpin the main upcoming progress in healthcare research and development. However, the biomolecules abundances in real samples can considerably alter the interface performance, where unwanted adsorption/adhesion can block signal response and significantly decrease the specificity of the assay. Herein, this review firstly offers a brief outline of several significances of fabricating high-sensitivity and low-background interfaces to adjust various targets' behaviors induced via bioactive molecules on the surface. Besides, some important strategies to resist non-specific protein adsorption and cell adhesion, followed by imperative categories of antifouling reagents utilized in the construction of high-performance solid sensory interfaces, are discussed. The next section specifically highlights the various nanocomposite probes based on antifouling-nanomaterials for electrode modification containing carbon nanomaterials, noble metal nanoparticles, magnetic nanoparticles, polymer, and silicon-based materials in terms of nanoparticles, rods, or porous materials through optical or chemical strategies. We specially outline those nanoprobes that are capable of identification in complex media or those using new constructions/methods. Finally, the necessity and requirements for future advances in this emerging field are also presented, followed by opportunities and challenges.

Concanavalin A-conjugated gold nanoparticle/silica quantum dot (AuNPs/SiQDs-Con A)-based platform as a fluorescent nanoprobe for the bioimaging of glycan-positive cancer cells.

The glycan receptor is a glycosylphosphatidylinositol glycoprotein that is overexpressed on the surface of various cancer cells and has been utilized for wide applications. In the present work, the surface of citrate-capped gold nanoparticles (cit-AuNPs) was modified with mercaptopropionic acid (MPA) molecules to provide carboxylic groups for secondary functionalization with amine anchored-silica quantum dots (Si-NH2 QDs) to produce cit-AuNPs-MPA/Si-NH2 QDs fluorescent nanoparticles. Concanavalin A (Con A) molecules were attached through thiol-AuNP bonds to produce the final cit-AuNPs/MPA/Si-NH2 QDs/Con A smart nanoparticles. The synthesized novel cit-AuNPs/MPA/Si-NH2 QDs/Con A nanoparticles were utilized for the bioimaging of glycan-overexpressed breast cancer cells. Fluorescence microscopy and flow cytometry results revealed that the cit-AuNPs/MPA/Si-NH2 QDs/Con A NPs can be efficiently taken up by cancer cells, with differentiating ability between overexpressed cancer cells and low-expressed normal cells. The cellular viability of the cit-AuNPs/MPA/Si-NH2 QDs/Con A NPs was tested by the MTT test, proving their biocompatible nature at the 200 µg mL-1 level. In conclusion, the fabricated cit-AuNPs/MPA/Si-NH2 QDs/Con A NPs could be utilized for the bioimaging of MCF-7 cancer cells even in the clinical setting after proper in vivo validation.

Quantification of methotrexate in plasma samples using highly fluorescent nanoparticles.

A novel amine-functionalized silica carbon polymer dots (APTES-CPDs)-based sensor was developed for methotrexate (MTX) detection. APTES-CPDs core-shell structure was synthesized via a one-step hydrothermal process and were employed as an optical sensor for MTX determination due to quenching effect of MTX on the fluorescence intensity of CPDs. Under optimum conditions, the developed sensor detects MTX in plasma samples in the dual linear ranges of 10 ng/mL to 2.0 µg/mL and 2.0 µg/mL to 50 µg/mL with a low limit of detections (LLOD)) of 10 ng/mL and 2.0 µg/mL, respectively. The obtained results are demonstrated the accuracy of the designed nanoprobe which is as high as to be used for the MTX detection in the routine therapeutic drug monitoring. In addition, the established nanoprobe shows significant specificity towards MTX compared to the co-administered drugs. Other advantages of this nanoprobe are its simple approach, high selectivity and sensitivity for MTX determination in plasma media which can approve the applicability of this MTX nanosensor in real samples.

A combination of amino-functionalized fibrous silica (KCC-1-NH2)/effectively and efficiently oxidized graphene oxide (EEGO) nanocomposite for dispersive solid-phase extraction, pre-concentration and fluorescence determination of total para-cresol in plasma samples of chronic kidney disease patients.

A novel amino-functionalized fibrous silica (KCC-1-NH2) and effectively and efficiently oxidized graphene oxide (EEGO) nanocomposite has been successfully synthesized. This nanocomposite was applied as a new sorbent in the dispersive solid-phase extraction (-SPE) to the preconcentration of total p-cresyl sulfate (pCS) in human plasma samples. The morphology and basic structure of the proposed nanocomposite were investigated through different techniques including field emission scanning electron microscopy (FESEM), energy dispersive X-ray (EDX), transmission electron microscopy (TEM), Fourier transform infrared (FTIR), and dynamic light scattering (DLS)/zeta potential techniques. The influence of different factors on the extraction efficiency, including the amount of sorbent, sample pH, extraction time, elution solvents and their volume, and desorption time were also investigated. The developed fluorescence-based method offers a linear dynamic range from 0.02 to 6 µg/mL with an acceptable correlation coefficient (R2 = 0.9982) and recovery (80%). The limit of quantification (LOQ) and limit of detection (LOD) were found to be 0.043 and 0.013 µg/mL, respectively. Plasma samples of five chronic kidney disease (CKD) patients were also analyzed and measured pCS concentrations were ranged from 16 to 41 µg/mL. The applicability of the method was successfully tested for the extraction and quantification of pCS from spiked and patients' plasma samples.