Quantitative Determination Research Articles

Advanced HPLC Method with Diode Array Detection Using a Phenyl-Bonded Column for Simultaneous Quantitation of Three Sunscreen Filters in a Moisturizing Sunscreen Cream for Acne-Prone Skin

This study introduces a novel, robust, and efficient method for the simultaneous quantitative determination of three sunscreen filters, namely, 4-methylbenzylidene camphor, octyl methoxycinnamate, and avobenzone, in a moisturizing sunscreen cream specifically designed for acne-prone skin. The method employs high-performance liquid chromatography with photodiode-array detection, providing a reliable separation of the analytes. Chromatographic separation was achieved using a Fortis Phenyl analytical column (150.0 × 2.1 mm, 5 μm), with isocratic elution at a flow rate of 0.4 mL/min. The mobile phase was composed of a 57/43 (v/v) mixture of acetonitrile/45 mM aqueous ammonium formate solution, ensuring sufficient resolution and peak symmetry for the target compounds. The method was validated comprehensively for critical performance parameters, including linearity, precision, accuracy, and robustness. Linearity was established across a suitable range for all three analytes, with high correlation coefficients. Precision was confirmed with intra-run and total precision coefficients of variation of ≤4.6%, while accuracy assessments yielded a percent recovery between 98.6 and 100.4, for all quality control levels. Additionally, the method was able to effectively separate the sunscreen filters from other cosmetic ingredients, such as [β-(1.3), (1.6)-D-glucan], low molecular weight (LMW) hyaluronic acid and plant extracts ensuring specificity in complex formulations. This straightforward and time efficient sample preparation process, involving methanol extraction followed by serial dilution, makes the method suitable for routine quality control in cosmetic laboratories. The method was successfully applied to the analysis of two different lots of a commercial sunscreen cream, achieving excellent recovery for all filters, ranging between 94.6% and 99.8%, thus demonstrating its reliability and applicability for the quality control of cosmetics.

Greenness and whiteness appraisal for bioanalysis of quetiapine, levodopa and carbidopa in spiked human plasma by high performance thin layer chromatography.

A sustainable HPTLC-densitometric method was developed for quantitative determination of Quetiapine (QUET), Levodopa (LD) and Carbidopa (CD) in presence of Dopamine (DOP) as an internal standard. This applicable technique was achieved by spiking human plasma and extraction was performed using the protein precipitation approach. The mobile phase used was acetone, dichloromethane, n-butanol, glacial acetic acid and water (3: 2.5: 2: 2: 1.75, by volume). Method validation was done according to US-FDA guidelines and was able to quantify Quetiapine, Levodopa and Carbidopa in the ranges of 100-4000, 200-8000 and 30-1300 ng/mL, respectively. Bioanalytical method validation parameters were assessed for the studied drugs. Finally, the analytical suggested methodology was evaluated using various green and white analytical chemistry metrics and other tools, such as the green solvent selection tool, analytical eco-scale, green analytical procedure index, analytical greenness metric approach and the red-green-blue algorithm tool. The results revealed that the applied analytical method had a minor impact on the environment and is a relatively greener option than other previously reported chromatographic methods.

Phytochemical, Mineral and Proximate Analysis of Herbal Tea Formulated for Malaria Management

The use of herbal tea preparations in the management of malaria infection has led to an increase in the production and research interest in teas. The readily available and cheap plant materials for production has made research efforts to be tailored towards development of different herbal tea preparations. In this study, the herbal tea, specially formulated for management of malaria infection, was subjected to various analysis. Qualitative and quantitative phytochemical determination indicated presence of saponin (9.71mg/100g), tannin (46.43mg/100g), phenols (904.01mg/100g), flavonoids (24.56mg/100g) and alkaloid (20.68mg/100g). The mineral component determination indicated that it was rich in Potassium (1.80 ppm), Magnesium (0.328 ppm) and Sodium (24.95ppm). Some heavy metals such as Cadmium (2.80ppm) and Iron (10.21ppm) were also detected. Proximate analysis carried out indicated moisture content (6.58%), Ash (9.23%), Carbohydrate (70.36%), Crude lipids (1.07%), Crude fibre (3.11%) and Crude protein (9.58%). Results shows that all the phytochemicals needed to fight malaria parasite are present in the herbal tea formulation. The mineral contents fall within World Health Organization acceptable level except for Cadmium and Iron.

An Alternative Method of Reversed-Phase HPLC for Identification and Quantitative Determination 0f Ergot Alkaloids

An Alternative Method of Reversed-Phase HPLC for Identification and Quantitative Determination 0f Ergot Alkaloids

Quantitative Determination of Volatile N-Nitrosamines in Meat Products Using Gas Chromatography Coupled to an Ion Mobility Spectrometer with an Ammonia Dopant.

Herein, we develop a novel method using gas chromatography-ion mobility spectrometry (IMS) with an ammonia dopant for the determination of volatile N-nitrosamines in meat products. The IMS system was implemented with Fourier deconvolution multiplexing to simultaneously improve the resolving power and sensitivity. The ammonia dopant mitigated the formation of N-nitrosamine dimer ions, suppressing the ionization of interferents and shifting the reactant peak from hydrated protons to hydrated ammonium ions. The ammonium adduct product ions of N-nitrosamines were confirmed using time-of-flight mass spectrometry. The instrument limits of detection and quantitation for nitrosamines were 0.78-1.79 and 2.60-5.82 ng/mL, respectively, with recoveries between 71.39% and 110.59% using a simple water distillation method. The developed method showed satisfactory performance when applied to the detection of nitrosamines in various meat products.

Research on the prediction method for fluvial-phase sandbody connectivity based on big data analysis--a case study of Bohai a oilfield

The connectivity of sandbodies is a key constraint to the exploration effectiveness of Bohai A Oilfield. Conventional connectivity studies often use methods such as seismic attribute fusion, while the development of contiguous composite sandbodies in this area makes it challenging to characterize connectivity changes with conventional seismic attributes. Aiming at the above problem in the Bohai A Oilfield, this study proposes a big data analysis method based on the Deep Forest algorithm to predict the sandbody connectivity. Firstly, by compiling the abundant exploration and development sandbodies data in the study area, typical sandbodies with reliable connectivity were selected. Then, sensitive seismic attribute were extracted to obtain training samples. Finally, based on the Deep Forest algorithm, mapping model between attribute combinations and sandbody connectivity was established through machine learning. This method achieves the first quantitative determination of the connectivity for continuous composite sandbodies in the Bohai Oilfield. Compared with conventional connectivity discrimination methods such as high-resolution processing and seismic attribute analysis, this method can combine the sandbody characteristics of the study area in the process of machine learning, and jointly judge connectivity by combining multiple seismic attributes. The study results show that this method has high accuracy and timeliness in predicting connectivity for continuous composite sandbodies. Applied to the Bohai A Oilfield, it successfully identified multiple sandbody connectivity relationships and provided strong support for the subsequent exploration potential assessment and well placement optimization. This method also provides a new idea and method for studying sandbody connectivity under similar complex geological conditions.

Sensitive determination of volatile nitrosamines with ambient pressure ammonium-adduct ionization mass spectrometry.

In recent years, the control of volatile N-nitrosamines (NAs) has been of interest in the pharmaceutical and food industries, as many of these compounds are probable human carcinogens. Thus, rapid and trace-level quantitative determination methods are in urgent demand. In this work, ambient pressure ammonium-adduct ionization mass spectrometry was proposed for the sensitive detection of volatile nitrosamines in various pharmaceutical headspaces. The ammonium ions produced through electrospray ionization acted as reactant ions for NAs to generate ammonium-NA adduct ions and underwent in-source collision-induced dissociation to produce protonated NAs, which were detected by mass spectrometry. The ionization selectivity and sensitivity for various volatile NAs were improved significantly using the developed method, which was demonstrated by the limit of quantification (LOQ) below 52ng L-1 for all NAs, and the quantitative performance was consequently improved. Different NAs exhibited almost equimolar response using NH4+ as the reactant ion, with at least a twofold enhancement in intensity for the individual compounds relative to when using H+ as the reactant ion. The proposed method is a rapid, sensitive, and environmentally economical approach that uses few reagents.

Quantitative Determination of Salivary Cariogenic Bacteria in Patients With Fixed Orthodontic Appliances.

Fixed orthodontic treatment can alter the oral environment, potentially increasing cariogenic bacteria levels. This study aimed to utilize real-time polymerase chain reaction (qPCR) to quantify salivary levels of cariogenic bacteria in patients undergoing orthodontic treatment with fixed appliances. This case-control study was conducted in Erbil, Iraq, from April 2024 to August 2024. One hundred participants were included in the study by random sampling method and equally divided into two groups: case group ororthodontic group and control group. The orthodontic group included participants with fixed orthodontic brackets, while the control group included participants without current or past orthodontic treatment with matching age and sex as the orthodontic group. Data were collected via questionnaires, clinical examinations, and saliva sampling. Salivary samples were analyzed using real-time PCR to detect Streptococcus mutans (SM) and Lactobacillus (LB) spp. levels. The age ranged from 18 to 30 years, and the mean age of participants was 23.96 ± 3.64 years. No significant differences were observed between the two groups across most parameters, except in the use of fluoride therapy, where the control group demonstrated better usage (p≤0.003). Real-time PCR results indicated a significant difference in the levels of LB and SM between patients with fixed orthodontics and the control group (p≤0.001). However, no significant differences were found in decay indices such as decayed, missing, and filled teeth; decayed, missing, and filled surfaces; and plaque index in this study. The quantity and levels of LB and SM were higher in the orthodontic group compared to the control group. Despite the elevated bacterial levels in the orthodontic group, there was no significant difference in decay and plaque indices between the two groups.

Investigation of ponatinib metabolism and drug-drug interactions with lycopene and shikonin in vitro and invivo

Ponatinib is approved for use in patients with chronic myeloid leukemia (CML) who are resistant to or intolerant to prior tyrosine kinase inhibitor (TKI) therapy. Given that ponatinib can induce significant cardiotoxicity when taken, and that most Chinese medicines have cardioprotective effects, it is possible to administer them in combination in clinic to alleviate adverse effects. The quantitative determination of ponatinib and its metabolite N-desmethyl ponatinib was optimized and fully verified by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). And the drug-drug interactions (DDI) of ponatinib with lycopene and shikonin, both in vivo and in vitro, were studied. The results of bioanalytical methodology showed that ponatinib and N-desmethyl ponatinib had good linearity in plasma samples, and their selectivity, accuracy, precision, stability, matrix effect and recovery were all satisfied with the need of quantitative analysis of samples. In animal experiments, compared with the control group, lycopene and shikonin significantly changed the pharmacokinetic parameters of ponatinib, including AUC(0-t), AUC(0-∞) and CLz/F, while having no effect on the pharmacokinetic parameters of N-desmethyl ponatinib. In vitro interaction studies indicated that lycopene showed mixed inhibition mechanism on ponatinib metabolism in both rat liver microsomes (RLM) and human liver microsomes (HLM). And, shikonin displayed mixed inhibition mechanism in RLM and competitive inhibition mechanism in HLM, respectively. In summary, the UPLC-MS/MS method can accurately and sensitively quantify ponatinib and N-desmethyl ponatinib, and provide further reference for clinical drug combination between ponatinib and lycopene or shikonin.

Cross-validation of methods for the quantitative determination of phenazepam and its active metabolite in human blood plasma at various extractions

Introduction. In conducting of therapeutic drug monitoring (TDM), often such situation arises where the drug concentration has measured by different methods or in different laboratories. To combine and analyze the data obtained with different methods, it is necessary to perform cross-validation procedure. Insufficient attention is paid to the statistical approaches used for this purpose.Aim. Performing cross-validation of different analytical methods for the quantitative determination of phenazepam (PHEN) and 3-hydroxyphenazepam (3-OH-PHEN) using the Bland – Altman analysis.Materials and methods. PHEN and 3-OH-PHEN concentrations in the blood plasma of patients (n = 100) with alcohol withdrawal syndrome were measured using high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS). The quantification of both analytes in each sample was measure twice by two different methods: solid phase extraction (SPE) and supported liquid extraction (SLE). Both methods have been fully validated before the experiment began. Cross-validation was performed at the end of the experiment using data from study samples. The Bland – Altman analysis was used to evaluate accuracy and precision. Deming regression was also used to identify a systematic error between measurement results.Results and discussion. The regression equations have been obtained between concentrations both analytes measured by different sample preparation methods. 95 % confidence intervals (CI) of the regression coefficients of both equations included one, and 95 % CI of the intercepts included zero. 95 % CI of the geometric mean of the individual SLE/SPE ratios was within the acceptable range (0.87; 1.15). These results confirm the absence of the influence of quantitative methods on the measurement of both analytes concentration. 66.7 % CI of the percent difference between two measurements was within acceptable limits (–0.2; 0.2), not exceeding 20 % of the range of their mean value. This confirms the acceptable precision between the methods. The estimated CIs were displayed in the Bland – Altman plots.Conclusion. The statistical approaches used in the work have confirmed the reproducibility of the results of different sample preparation methods. In addition to cross-validation, the statistical algorithm from this paper using Bland – Altman analysis can be successfully employed to assess accuracy and precision during bioanalytical method validation and evaluation of the acceptance of analytical runs, as well as to determine the level of reproducibility of incurred samples.

Dynamic Constitutive Model of Tungsten‐Whisker‐Reinforced Aluminum/Polytetrafluoroethylene Composite Material and Quantitative Determination of Impact Reaction Release Energy in Vacuum Environment

To meet the strength requirements of aluminum/polytetrafluoroethylene (Al/PTFE) in practical applications, tungsten (W) whisker is added into the traditional formula Al/PTFE (26.5%/73.5%) to improve the dynamic compressive strength of the reactive material. Mechanical properties of reactive material specimens prepared by sintering are tested. In the results, it is shown that the dynamic compressive strength of Al/PTFE‐reactive material reinforced by tungsten whisker with volume fraction of 13.1% can reach 141 MPa at strain rate of 1800 s−1. The constitutive model of dynamic compression for tungsten‐whisker‐enhanced Al/PTFE‐reactive material is constructed, and the results are basically consistent with the experimental results. The quantitative impact energy release of reactive material specimens under vacuum environment are performed by using the two‐stage light gas gun loading system, and the quantitative impact release energies of Al/PTFE‐reactive material reinforced by tungsten whisker under different whisker percentage content and diameter are obtained.

Two-dimensional photonic crystal acetylcholinesterase hydrogel and organohydrogel sensors for efficient detection of organophosphorus compounds

Sensors capable of detecting organophosphorus (OP) compounds have attracted the most attention owing to severe OP contamination worldwide. Despite many years of research, the developed OP sensors mainly focused on detecting water-soluble OPs in proper environments and the exploration of OP sensors suitable in resource-limited areas is extremely challenging. Here, a simple two-dimensional photonic crystal (2D PC) hydrogel featuring capabilities of effectively quantitative determination of OP compounds is facilely constructed by immobilizing the enzyme acetylcholinesterase (AChE) onto a bovine serum albumin (BSA) protein hydrogel. Owing to the specific interaction between AChE and OP compounds, the OP compounds are easily bound to the hydrogel, triggering volume phase transition and resulting in apparent Debye diffraction ring variations. The resulting hydrogel sensors show a limit of detection (LoD) of 2.23 nM for trichlorfon and 0.07 nM for diethyl methylphosphonate (DMPP), respectively. On the basis of the hydrogel, a responsive organohydrogel is facilely fabricated utilizing a solvent exchange strategy to meet the requirements of applications in harsh environments and detection of the non-water-soluble OP compounds. The organohydrogel sensors, however, demonstrated a LoD of 0.70 μM for trichlorfon and 4.46 μM for DMPP, respectively. This work provides new light on the development of next-generation stable, low-cost, and portable field sensing devices.

Experimental constraints on serpentinite carbonation in the presence of a H2O–CO2–NaCl fluid

Serpentinite carbonation contributes to the deep carbon (C) cycle. Recently, geophysical and numerical studies have inferred considerable hydrothermal alteration in plate bending faults, opening the possibility of significant C storage in the slab mantle. However, there is a lack of quantitative determination of C uptake in serpentinized mantle rocks. Here, we experimentally constrain serpentinite carbonation in H2O–CO2–NaCl fluids to estimate C uptake in hydrated mantle rocks. We find that serpentinite carbonation results in the formation of talc and magnesite along the serpentinite surface. The presence of porous reaction zones (49.2% porosity) promotes the progress of carbonation reactions through a continuous supply of CO2-bearing fluids to the reaction front. Added NaCl effectively decreases the serpentinite carbonation efficiency, particularly at low salinities (< 5.0 wt%), which is likely attributed to the reduction in fluid pH and the transport rate of reactants, and the increase in magnesite solubility. Based on previous and our experiments, we fit an empirical equation for the reaction rate of serpentinite carbonation. Extrapolation of this equation to depths of plate bending fault systems suggests that serpentinite carbonation may contribute to an influx of up to 7.3–28.5 Mt C/yr in subduction zones. Our results provide new insights into serpentinite carbonation in environments with high fluid salinities and potentially contribute to the understanding of the C cycle in subduction zones.

Plasmonic Coacervate as a Droplet-Based SERS Platform for Rapid Enrichment and Microanalysis of Hydrophobic Payloads.

A novel and simple coacervate microdroplet-based detection platform for the quantification of trace hydrophobic analytes is presented. Herein, taking advantage of the effective encapsulation and enrichment performance of the condensed coacervates, plasmonic metallic silver nanoparticles (AgNPs) and target hydrophobic analytes are simultaneously concentrated into a single microdroplet. The coencapsulation of AgNPs within coacervates promotes the formation of aggregates with a lot of "hot spots" for surface-enhanced Raman scattering (SERS) enhancement, facilitating the sensitive analysis of hydrophobic analytes by SERS technology. Such plasmonic coacervates are easily prepared and exhibit good reproducibility and signal uniformity. Optimized SERS performance by modulating the volume of encapsulated AgNPs enables quantitative determination of hydrophobic analytes of Nile Red, chlorpyrifos, benzo[e]pyrene, 20 and 50 nm polystyrene nanoplastics with low detection limits of 10-12 M, 10-9 M, 10-10 M, 0.05 ppb, and 0.5 ppb, and an approximately linear correlation between SERS signals and the analytical concentrations. This study opens a new convenient SERS platform for the ultrasensitive detection of hydrophobic hazardous substances, potentially becoming a rapid analysis method for extensive applications ranging from food safety to environment monitoring.

Bismuth-coated screen-printed carbon electrodes for quantitative voltammetric determination of formaldehyde in urotropin

For the first time, a bismuth-modified electrode is used for the quantitative voltammetric determination of formaldehyde (FA) in urotropin. The well-established method of forming formaldehyde hydrazone (FAH) in the presence of hydrazine sulphate (HRZ) was employed to transform the hydrated form of FA into its electrochemically active derivative. The reduction current of FAH was recorded in a phosphate buffer solution (PBS, pH 5.2±0.1) in the presence of 0.09 M HRZ on a screen-printed carbon electrode that had been preliminarily modified with a bismuth film. A 20-second electrolysis at an applied potential of –0.28 V in a solution comprising 1 M HCl, 0.5 M NaBr, and 4 g/L Bi(III) resulted in the formation of a well-developed metal layer with an active surface. The determination of FA in technical urotropin yielded satisfactory results, indicating the potential for further applications of Bi/SPCE in pharmaceutical analysis.

Unraveling the Source of Self-Induced Diastereomeric Anisochronism in Chiral Dipeptides.

Mastering of analytical methods for accurate quantitative determinations of enantiomeric excess is a crucial aspect in asymmetric catalysis, chiral synthesis, and pharmaceutical applications. In this context, the phenomenon of Self-Induced Diastereomeric Anisochronism (SIDA) can be exploited in NMR spectroscopy for accurate determinations of enantiomeric composition, without using a chiral auxiliary that could interfere with the spectroscopic investigation. This phenomenon can be particularly useful for improving the quantitative analysis of mixtures with low enantiomeric excesses, where direct integration of signals can be tricky. Here, we describe a novel analysis protocol to correctly determine the enantiomeric composition of scalemic mixtures and investigate the thermodynamic and stereochemical features at the basis of SIDA. Dipeptide derivatives were chosen as substrates for this study, given their central role in drug design. By integrating the experiments with a conformational stochastic search that includes entropic contributions, we provide valuable information on the dimerization thermodynamics, the nature of non-covalent interactions leading to self-association, and the differences in the chemical environment responsible for the anisochronism, highlighting the importance of different stereochemical arrangement and tight association for the distinction between homochiral and heterochiral adducts. An important role played by the counterion was pointed out by computational studies.

The molecular design of novel phospholipid-inspired ionic liquid transdermal penetration enhancers: Innovative insights on the action mode and mechanism

Ionic liquid transdermal penetration enhancers (IL@TPEs) as new enhancement methods have significant advantages in the transdermal drug delivery system. However, the scientific frameworks for the design of efficient IL@TPEs and their applications in transdermal formulations were still lack. So, a series of novel biomimetic phospholipid-inspired IL@TPEs (PIL@TPEs) were designed and synthesized. The developed QSARs proved that enhancement efficacy of PIL@TPEs depended on pKa of drugs and M.W., Polar., and pKa of cations. Surprisingly, the PIL@TPEs dissociated during transdermal process, and skin penetration amounts of acidic drugs was inversely proportional to skin retention amounts of cations, which showed that action modes of PIL@TPEs were different from conventional enhancers. The novel mechanisms of PIL@TPEs were elucidated by quantitative determination of dynamic interaction among cations, anions, drugs, and skins. The PIL@TPEs with high enhancement efficiency owned strong interactions with drugs determined by ATR-FTIR, Raman and NOESY. Moreover, the PIL@TPEs owning better stability in skin ensured the production of strong interactions with lipids and keratins characterized by ATR-FTIR, 1H NMR and CLSM. The good safety of optimized PIL@TPEs was proved by determining cytotoxicity, apoptosis, inflammatory cells, and cytokines. In conclusion, this project will make an important contribution to the design and application of IL@TPEs.

The method of capillary electrophoresis for quantitative determination of hydrophobized hyaluronic acid in its micellar forms

Micelles based on hydrophobized hyaluronic acid (HA) are frequently used in targeted drug delivery systems. Capillary zone electrophoresis (CZE) was utilized for the quantitative determination of hydrophobized and native HA. A universal methodology was developed, suitable for the quantitative analysis of amphiphilic derivatives of hyaluronan (oleyl hyaluronan and hyaluronan conjugate with naphthalimide fluorophore) and native HA with varying molecular weights (15, 150, and 800 kDa). Furthermore, methodologies were proposed for the simultaneous quantification of a drug substance and oleyl hyaluronan in micellar forms based on the latter. The CE technique was applied for analyzing oleyl-hyaluronan-based micellar forms of two poorly soluble drug substances with oppositely charged ionic forms (loperamide and rifabutin). The examples contained in the study demonstrate a range of analytical sensitivity (LOD) for hyaluronan from 11 to 40 μg/mL and for the drug substance from 0.4 to 0.6 μg/mL. The study also showcases the accurate quantitative determination of rifabutin and loperamide in oleyl-hyaluronan-based micellar forms without the need for sample preparation. Thus, the proposed methodologies can be used to quantify native HA or its amphiphilic derivatives and simultaneously determine drug substances of various nature.

Superwettable microchip for visual detection of date-rape drug in beverages

Gamma-hydroxybutyric acid (GHB), which is used in drug-facilitated sexual assault, is typically delivered in the form of gamma-butyrolactone (GBL) in beverage carriers. In response to the growing need of methods for rapid and on-site sensing of the drugs, we carried out an investigation aimed at developing a superwettable microchip for visual detection of the date-rape drug GBL in adulterated beverages. The superwettable microchip is comprised of Fe3O4 nanoparticles which aggregate to form a reactive spot on a superhydrophobic fabric. The spot captures and confines microdroplets containing GBL on the superhydrophobic substrate. Fe(III), generated by HCl promoted reaction of the Fe3O4 spot, then participates in a colorimetric reaction with GBL to form a naked eye observable colored complex. Quantitative determination of GBL in beverages is achieved by analyzing the red proportion of a smartphone camera derived image. The detection range of this assay protocol is ca. 0.1–10 mg mL−1, which meets the requirements to detect potentially dangerous concentration of GBL in drinks. The detection stability of the superwettable microchip for GBL in water and common beverages is maintained over at least 30 cycles.

A-084 Performance evaluation of Sentinel Diagnostics’ Ammonia Ultra assay on Beckman Coulter AU/DxC AU Systems

Abstract Background The analytical performances of the Ammonia Ultra assay for in vitro quantitative enzymatic determination of Ammonia in human plasma have been evaluated on Beckman AU/DxC AU Systems. Ammonia, derived from the amino acids catabolism and from the action of intestinal bacteria on dietary proteins, is converted to non-toxic urea by the liver. An excess of ammonia can have toxic effects on the Central Nervous System with possible neurological disturbances. Elevated ammonia levels can also be observed in severe liver failure as may occur in Reye’s syndrome, viral hepatitis or cirrhosis. Methods Ammonia, in the presence of glutamate dehydrogenase (GLDH), combines with alpha-ketoglutarate and NADH to yield glutamate and NAD+. The decrease in 340 nm absorbance (NADH → NAD+) is proportional to the ammonia level in the examined plasma. The reagent contains excess of lactate dehydrogenase (LDH) to rapidly reduce endogenous pyruvate and avoid its interference with the assay system. Performances evaluation have been done following current CLSI guidelines protocols. Results 3 calibrator lots and 2 reagent lots have been used for testing. For each test and among different combinations of reagents and calibrators, worst case has been reported in the table. Conclusions Analytical performances of Ammonia Ultra assay on Beckman AU/DxC AU Systems meet the requirements for its use as quantitative determination of Ammonia in human plasma and make this assay very suitable for the routine measurement of this analyte.