Simple and sensitive determination of sparfloxacin in pharmaceuticals and biological samples by immunoassay

Plasma quinolone concentrations are not routinely measured in clinical practice.However,in order to optimize quinolone treatment,monitoring of plasma concentrations could sometimes be useful particularly in critically ill patients.In this study,anti-sparfloxacin antibody was obtained by immunizing rabbits with sparfloxacin conjugated with bovine serum albumin using isobutyl chloroformate method.After the assay procedure was optimized,the standard curve of sparfloxacin was established.The practical measuring range of the competitive ELISA extended from 5 ng/mL to 2 mg/mL.The recovery rates and coefficients of variation for rat plasma,urine and tissues were 87.7-106.2% and 4.8-15.3%,respectively.To demonstrate the potential of the ELISA,a preliminary pharmacokinetics and tissue distribution study of sparfloxacin in rats and quantitative analysis of sparfloxacin in several pharmaceuticals were performed and compared with high-performance liquid chromatography(HPLC).The experimental data indicated that the proposed method would be a valuable tool in therapeutic drug monitoring(TDM) for sparfloxacin.

Fabrication of edge-curled petals-like covalent organic frameworks and their properties for extracting indole alkaloids from complex biological samples

In this study,a functionalized covalent-organic framework(COF)was first synthesized using porphyrin as the fabrication unit and showed an edge-curled,petal-like and well-ordered structure.The synthesized COF was then introduced to prepare porous organic polymer monolithic materials(POPMs).Two composite POPM/COF monolithic materials with rod shapes,referred to as sorbent A and sorbent B,were prepared in stainless steel tubes using different monomers.Sorbents A and B exhibited relatively uniform porous structures and enhanced specific surface areas of 153.14 m;/g and 80.01 m;/g,respectively.The prepared composite monoliths were used as in-tube solid-phase extraction(SPE)sorbents combined with HPLC for the on-line extraction and quantitative analytical systems.Indole alkaloids(from Catharanthus roseus G.Don and Uncaria rhynchophylla(Miq.)Miq.Ex Havil.)contained in mouse plasma were extracted and quantitatively analyzed using the online system.The two composite multifunctional monoliths showed excellent clean-up ability for complex biological matrices,as well as superior selectivity for target indole alkaloids.Method validation showed that the RSD values of the repeatability(n=6)were≤3.46%,and the accuracy expressed by the spiked recoveries was in the ranges of 99.38%-100.91%and 96.39%-103.50%for vinca alkaloids and Uncaria alkaloids,respectively.Furthermore,sorbents A and B exhibited strong reusability,with RSD values≤5.32%,which were based on the peak area of the corresponding alkaloids with more than 100 injections.These results indicate that the composite POPM/COF rod-shaped monoliths are promising media as SPE sorbents for extracting trace compounds in complex biological samples.

Determination of Trace Rare Earth Elements in Biological Samples by ICP-AES with Tungsten-spiral Electrothermal Vaporization Sampling

A method for determination of trace rare earth elements in biological sample by ICP-AES with tungsten-spiral electrothermal vaporization(TETV-ICP-AES)sampling has been developed in this work. Several influencing factors including desolvation and vaporization parameters,carrier gas flow rate and ma- trix effect were investigated in detail.Under optimal experimental conditions,the detection limits for La,Nd, Gd,Dy,Ho,Yb,Lu and Y were obtained in the range of 10^(-9)～10^(-11) g,and they are comparable to and,in most instances,better than those for the GFAAS and conventional pneumatic nebulisation-ICP-AES.The precision(RSD)obtained for this method is less than 6%.

Study of Direct Determination of Trace Lanthanum in Biological Samples by ICP-AES Combined with Fluorination Electrothermal Vaporization Technique

A new method for direct determination of lanthanum in solid biological materials by fluorination electrothermal vaporization ICP-AES technique with polytetrafluoroethylene(PTFE) disperser as a fluorination agent has been described. The effect of particle size on the signal intensity of La has been investigated. The vaporization behaviour of lanthanum and the main factors affecting fluorinating vaporization have been observed.Under optimum experimental conditions,the detection limit of La to this method is 2.0 ng/ml,and the RSD is 4.5%.The proposed method has been applied to determining directly trace lanthanum in solid biological standard reference materials without any chemical pretreatment,and the determined values are in good agreement with the certified ones.

THE USE OF CHELATING RESIN FOR THE PRE-CONCENTRATION OF TRACE RARE EARTH ELEMENTS IN BIOLOGICAL SAMPLES AND THEIR DETERMINATION

Macroporous amino-carboxvlic chelating resin of acrylic acid series may concentrate trace rare earth in the presence of 0.1 mol/l α-hydroxy-isobutyric acid at pH 4.5.With 0.10 mol/l HCl as eluant rare earth may be eluted quantitatively and separated from Ca,Mg,Fe and Cu.This method is used to determine trace amount of rare earth in human hair,stone (in kidney and bladder),peanut,grape,strawberry,cucumber,pig liver and other biological samples.

Interaction between Cadmium and Zinc Levels in the Biological Samples of Type 1 Diabetic Mellitus Children, Reside in Different Areas of Sindh, Pakistan

Type 1 Diabetes mellitus (T1DM) is one of the familiar childhood immune-mediated onsets and can lead to early mortalities and morbidities. It can arise at any stage, but the peak of occurrence is reported less than 18 years of age. T1DM cases in Pakistan were less than 2% of the total diabetic population. The current work designed to assess the concentration of cadmium (Cd) and zinc (Zn) in blood, scalp hair and serum samples of T1DM children, age ranged 1 - 14 years of both genders. For comparison purpose, the age-matched referent subjects of both genders were tested. The microwave-assisted acid digestion procedure was used to determine the elemental analysis in the biological samples of T1DM children and referent subjects. The resulted data of certified reference material of blood, scalp hair, and serum validated the certainty of the designed method. The analysis of Zn was performed by flame atomic absorption spectrometry, while the Cd contents were determined by electrothermal atomic absorption spectrometry. T1DM affected children of both genders have lower Zn level in the blood, scalp hair, and serum samples. Whereas, the levels of Cd were found to be higher in the biological samples of T1DM affected children as compared to referent subjects. The finding of the current study is a significant hypothesis for medical experts, to diagnose the deficiency of essential (Zn) and toxicity of heavy/toxic element (Cd) in the biological specimen of T1DM affected children.

Calcium and Lead Levels in the Biological Samples and Their Effect on the Biochemical Parameters of Indoor and Outdoor Workers of Five Zonal Areas of Coal Mining Field

Workers of the Lakhra coal mining Sindh Pakistan were affected by a multi-factorial exposure of heavy metals. The prevalence of lead (Pb) exposures in work-related symptoms among workers of indoor and outdoor mining of Lakhra coal mining was evaluated in detail. The biological samples (scalp hair and blood) of 270 workers of Lakhra coal mining {official and control subjects} (indoor and outdoor) 18 to 55 years age group and 70 referent subjects of same age-groups were examined. The biological samples (blood and scalp hair) of workers of Lakhra coal mining and referent subjects were analyzed for calcium (Ca) and lead (Pb). Flame atomic absorption spectrometry was applied for the determination of the Ca in digested samples of studied subjects by the microwave oven whereas, the Pb contents were measured by electrothermal atomic absorption spectrometry. The validity of the methodology was checked by using the certified human hair material (BCR 397) and lyophilized human blood (Clincheck). The results indicated the higher contents of Pb in biological samples of indoor and outdoor mining workers than referents. However, the levels of Ca were found to be lower in indoor and outdoor mining workers than those of referents. It can be concluded that immediate action should be taken by recommended authorities to improve the ventilation and hygiene practices inside the industries, mines, and other workplaces.

Elemental Concentrations in Biological Samples of Coronavirus Disease (COVID-19) and Other Pulmonary Disease Patients

Background: Recently, Coronavirus Disease 2019 (COVID-19) has been affected by the extreme acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and has developed important global health anxiety. At this time, the treatment options for this disease are only moderately successful. Smoking has been related to COVID-19 and its mortality, and it has the potential to affect bacteriological and viral conversion, clinical effects, and treatment outcomes. Methods: The aim of this study was to determine the relationship between critical essential trace {zinc (Zn), iron (Fe), and copper (Cu)} and toxic {cadmium (Cd) and nickel (Ni)} elements in human biological samples such as scalp hair, serum, saliva, blood, nasal fluid, and sputum of smoking and nonsmoking male COVID-19 patients (n = 139, age range 25 - 38 years) from Hyderabad, Pakistan. For comparative purposes, the biological specimens of referent persons (n = 83), pulmonary Tuberculosis (T.B) patients (n = 67), Pneumonia (n = 56) of the same age groups were also be analyzed. Microwave oven mediated digestion method was employed, and digestion of samples was carried out with the help of 2:1 mixture solution of HNO3 (65%) and H2O2 (30%). Atomic absorption spectrometry was employed for the determination of elemental concentrations from the microwave oven employed digested samples. Results: The found average of essential elements (Zn, Fe, and Cu) in biological specimens of smoker and nonsmoker male COVID-19, T.B, and Pneumonia patients was found to lower, whilst cadmium and nickel were found to be higher when compared with samples from referents (p < 0.001). Conclusions: Improved elemental (Cu, Fe, Zn) concentrations may also decrease the risk of bacterial co-infection by enhancing the mucociliary clearance and respiratory epithelial barrier function, in addition to providing direct antibacterial effects against S. pneumoniae. Our findings also suggest that higher Cd and Ni concentrations are linked to cigarette smoking, which could lead to COVID-19 and other lung-infected diseased recurrences. However, further clinical and experimental research is required.

ANALYSIS OF STRYCHNINE ALKALOIDS IN BIOLOGICAL SAMPLES BY THIN-LAYER CHROMATOGRAPHIC DENSITOMETRY AND DISTRIBUTION STUDY OF STRYCHNINE IN INTOXICATED RATS

A sensitive and rapid analytical procedure was established to detrmine strychnie alkaloids in diverse biological specimens. Samples were treated with hydrochloric acid to precipitate protein and scanned in thin-layer chromatograpic scanner in zig-zag dual-wavelength reflection mode.Detection sensitivity for strychniue was about 0. 05 μg. Quantification recoveries were between89. 2% and 99. 5% with RSD between 2. 92% and 13. 21 %. The method was applied to investigate thedistribution or strychnine in intoxicated rats. The results showed that there were remarkable difference in organs or tissues with the aigaest concentration 4. 56±1. 41μg/g in liver. Diversification experiment proves the method also could be used to analyze other alkaloids in biological samples if theconditions slightly modified.

Analysis of Fatty Acid in Biological Samples Using Liquid Chromatography–Quadrupole-Orbitrap Mass Spectrometry Under Parallel Reaction Monitoring Mode

Fatty acids(FAs)are an important class of functional small molecules and participate in a variety of life biological processes.This experiment proposed a liquid chromatography–quadrupole-Qrbitrap mass spectrometry(LC–Q-Orbitrap MS)method to perform FA profiling under parallel reaction monitoring(PRM)acquisition mode.The FA was derivatized by 2-dimethylaminoethylamine(DMED)to increase the ionization e fficiency and provide the characterized fragment pattern.The mass spectra obtained from full scan,multiple ion monitoring(MIM),and PRM were compared.The results showed that the protonated ion of FA+DMED–H2O was detected and the neutral loss of 45.06 Da was observed in the tandem mass spectrum.The PRM method provided the highest selectivity and sensitivity for FA detection with the help of accurate mass weight and specific neutral loss-based fragments.The method validation was performed using the FA standards and pooled quality control serum sample,which showed that the established method had good repeatability,stability,and linearity.Finally,the developed method was successfully applied to analyze the rat serum and brain tissue samples for the drug e ffi-cacy evaluation of gross saponin of Tribulus terrestris L.fruit(GSTTF)against middle cerebral artery occlusion(MCAO)-induced ischemic stroke.This method has extensive practicability and great potential in the detection of FA,especially for the analysis of samples with complex matrices.

New techniques of on-line biological sample processing and their application in the field of biopharmaceutical analysis

Biological sample pretreatment is an important step in biological sample analysis. Due to the diversity of biological matrices, the analysis of target substances in these samples presents significant challenges to sample processing. To meet these emerging demands on biopharmaceutical analysis, this paper summarizes several new techniques of on-line biological sample processing: solid phase extraction, solid phase micro-extraction, column switching, limited intake filler, molecularly imprinted solid phase extraction, tubular column, and micro-dialysis. We describe new developments, principles, and characteristics of these techniques, and the application of liquid chromatography mass spectrometry (LC MS) in biopharmaceutical analysis with these new techniques in on-line biological sample processing. (C) 2016 Chinese Pharmaceutical Association and Institute of Materia Medica, Chinese Academy of Medical Sciences. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND

A new kinetic spectrophotometric method for the determination of major metabolite of heroin in biological samples

This study describes a simple,rapid and selective catalytic kinetic spectrophotometric method for the determination of 6- monoacetylmorphine（6-MAM） as major metabolite of heroin in biological samples.The method is based upon the catalytic effect of 6-MAM on the oxidation of Janus Green by bromate in acid media.The reaction was followed spectrophotometrically by measuring the decrease in absorbance of Janus Green at 618 nm.The dependence of sensitivity on the reaction variables was studied.Under optimum conditions,two linear calibration curves over the range 0.1-1.0μg mL^（-1） and 1.0-34.0μg mL^（-1） of 6- MAM were obtained.The detection limit was 1.2×10~2μg mL^（-1） of 6-MAM.The relative standard deviations for six replicate determinations of 0.8 and 5.0μg mL^（-1） of 6-MAM were 1.4 and 1.1%respectively.The effect of various species commonly associated with heroin in real samples was also investigated.The proposed method was successfully applied in human urine and serum samples with satisfactory results.

A highly selective and instantaneous upconversion fluorescent nanoprobe for ascorbic acid detection in biological samples

Ascorbic acid（AA） serves as a key coenzyme in many metabolic pathways. Enough daily AA supplements from different dietary sources are the only way for human to maintain their AA levels in body.Determination of AA content in different foods guides to build healthy diet, which is of great biomedical significance. Hence, developing a highly selective and instantaneous fluorescent nanoprobe for the detection of AA in biological samples is highly needed. Here we present a novel turn-on fluorescent nanoprobe using lanthanide-doped upconversion nanoparticles（UCNPs） and cobalt oxyhydroxide（Co OOH） nanoflakes for monitoring AA in fruit samples. In this nanosystem, the UCNPs can be adsorbed onto the Co OOH nanoflakes, leading to a remarkable fluorescence decrease through Fo？rster resonance energy transfer. Furthermore, the AA could trigger the disassembly of the Co OOH to liberate the upconverted fluorescence. The UCNPs-based nanoprobe can provide an effective platform for highly selective and rapid detection of AA in biological samples.

Ultrasound-Assisted Low-Density Solvent Dispersive Liquid-Liquid Extraction for the Determination of Amphetamines in Biological Samples Using Gas Chromatography-Mass Spectrometry

In order to control drug crime effectively,it is necessary to develop selective analytical methods suitable for unambiguous identification and determination of drugs in illicit samples and biological matrices.A novel microextraction technique based on ultrasound-assisted low-density solvent dispersive liquid-liquid microextraction,(UA-LDS-DLLME)has been applied to the determination of four amphetamines(methamphetamine,amphetamine,3,4-methylenedioxymethamphetamine,and 3,4-methylenedioxyamphetamine)in urine samples by gas chromatography-mass spectrometry.The parameters affecting extraction efficiency have been investigated and optimized.UA-LDS-DLLME used ultrasound energy to assist in the emulsification process without any disperser solvent.Under the optimized conditions,linearity was observed for all analytes in the 0.15-10 p,g/mL range with correlation coefficients(R)ranging from 0.9886 to 0.9894.The recoveries of75.6-91.4%with relative standard deviations of 2.5-4.0%were obtained.The limits of detection(S/N=3)were estimated to be in the 5-10 ng/mL range.The UA-LDS-DLLME technique had the advantages of shorter extraction time,suitability for simultaneous pretreatments of batches of samples,and the higher extraction efficiency.It was successfully applied to the analysis of amphetamines in real human urine samples.

Multi-Compartment SCFA Quantification in Human

Short-chain fatty acids (SCFA) play an important role in human biochemistry. They originate primarily from the digestive system through carbohydrates microbial fermentation. Most SCFA produced in the colon are absorbed by the intestinal wall and enter the bloodstream to be distributed throughout the body for multiple purposes. At the intestinal level, SCFA play a role in controlling fat storage and fatty acid metabolism. The effects of these beneficial compounds therefore concern overall health. They facilitate energy expenditure and are valuable allies in the fight against obesity and diabetes. SCFA are also involved in the regulation of the levels of several neurotransmitters such as GABA (γ-aminobutyric acid), glutamate, serotonin, dopamine, and norepinephrine. Their role is also highlighted in many inflammatory and neurodegenerative diseases such as Alzheimer’s disease (AD) or Parkinson’s disease (PD). To have a realistic picture of the distribution of SCFA in different biological compartments of the human body, we propose to study SCFA simultaneously in five human biological samples: feces, saliva, serum, cerebrospinal fluid (CSF), and urine, as well as in Dried Blood Spot (DBS). To evaluate their concentration and repeatability, we used 10 aliquots from pooled samples, analyzed by 3-nitrophenylhydrazine (3-NPH) derivation and liquid chromatography coupled with high sensitivity mass spectrometry (LC-QqQ-MS). We also evaluated the SCFA assay on Dried Blood Spot (DBS). In this work, we adapted the pre-analytical parts for each sample to be able to use a common calibration curve, thus facilitating multi-assay quantification studies and so being less time-consuming. Moreover, we proposed new daughter ions from the same neutral loss (43 Da) to quantify SCFAs, thus improving the sensitivity. In conclusion, our methodology, based on a unique calibration curve for all samples for each SCFA, is well-suited to quantified them in a clinical context.

Accurate and sensitive determination of hydroxychloroquine sulfate used on COVID-19 patients in human urine,serum and saliva samples by GC-MS

A rapid,accurate,and sensitive analytical method,ultrasonication-assisted spraying based fine droplet formationeliquid phase microextractionegas chromatographyemass spectrometry(UA-SFDF-LPME-GCMS),was proposed for the determination of trace amounts of hydroxychloroquine sulfate in human serum,urine,and saliva samples.To determine the best extraction strategy,several liquid and solid phase extraction methods were investigated for their efficiencies in isolation and preconcentration of hydroxychloroquine sulfate from biological matrices.The UA-SFDF-LPME method was determined to be the best extraction method as it was operationally simple and provided accurate results.Variables such as the extraction solvent,spraying number,sodium hydroxide concentration and volume,sample volume,mixing method,and mixing period were optimized for the proposed method using the onevariable-at-a-time approach.In addition,Tukey’s method based on a post hoc comparison test was employed to evaluate the significant difference between the parameters inspected.After the optimization studies,the limit of detection(LOD)and limit of quantification(LOQ)were determined to be 0.7 and 2.4 mg/kg,respectively.The sensitivity of the GC-MS system based on the LOD was enhanced approximately 440-fold when the UA-SFDF-LPME method was employed.Spiking experiments were also conducted for the human serum,urine,and saliva samples to determine the applicability and accuracy of the proposed method.Recoveries for the human serum,urine,and saliva samples were found to be in the ranges of 93.9%-101.7%,95.2%-105.0%,and 93.1%-102.3%,respectively.These results were satisfactory and indicated that the hydroxychloroquine sulfate level in the above biological samples could be analyzed using the proposed method.

Balloon-based exposed payload designed for astrobiological research in Earth’s near space

Earth’s near space,located in the region between 20 and 100 km above sea level,is characterized by extreme conditions,such as low temperature,low atmospheric pressure,harsh radiation,and extreme dryness.These conditions are analogous to those found on the surface of Mars and in the atmosphere of Venus,making Earth’s near space a unique natural laboratory for astrobiological research.To address essential astrobiological questions,teams from the Chinese Academy of Sciences(CAS)have developed a scientific balloon platform,the CAS Balloon-Borne Astrobiology Platform(CAS-BAP),to study the effects of near space environmental conditions on the biology and survival strategies of representative organisms in this terrestrial analog.Here,we describe the versatile Biological Samples Exposure Payload(BIOSEP)loaded on the CAS-BAP with respect to its structure and function.The primary function of BIOSEP is to expose appropriate biological specimens to the harsh conditions of near space and subsequently return the exposed samples to laboratories for further analysis.Four successful flight missions in near space from 2019 to 2021 have demonstrated the high reliability and efficiency of the payload in communicating between hardware and software units,recording environmental data,exposing sample containers,protecting samples from external contamination,and recovering samples.Understanding the effects of Earth’s near space conditions on biological specimens will provide valuable insights into the survival strategies of organisms in extreme environments and the search for life beyond Earth.The development of BIOSEP and associated biological exposure experiments will enhance our understanding of the potential for life on Mars and the habitability of the atmospheric regions of other planets in the solar system and beyond.

色质联用技术在毒物分析中的应用

从生物样品中检测未知物,一直以来是毒物分析中的一个难点.如何有效地提取目标化合物,并选择适合的检测手段将未知物筛选出来,GC-MS方法是快速筛选多种毒物和药物的一个简便、可靠的手段.随着MS技术的不断发展,更灵敏、更准确的特性将使其成为药物和毒物分析中必不可少的一门技术.……

生物检材中氧乐果及其分解物的GC-MS分析

A method was established for the analysis of omethoate and it’s concomitants and decomposed product in biological samples by GC-MS.This method is rapid,reliable and accurate.

Phthalate Exposure and Human Semen Quality in Shanghai:A Cross-sectional Study

Objective To monitor the level of phthalates in human semen samples and to analyze the relationship between phthalate levels and semen parameters. Methods Concentrations of three kinds of commonly used phthalates （di-ethyl phthalate, DEP; di-n-butyl phthalate, DBP; di-2-ethylhexyl phthalate, DEHP） were measured using reversed-phase HPLC. Semen parameters were measured by computer aided sperm analysis （CASA）. Results The three phthalates were detected in most of the biological samples, with median levels of 0.30 mg/L （0.08-1.32 mg/L） in semen specimens. There was a significant positive association between liquefied time of semen and phthalate concentrations of semen. The correlation coefficient was 0.456 for DEP, 0.475 for DBP, and 0.457 for DEHP, respectively. There was no significant difference between phthalate concentrations of semen and sperm density or livability, though the correlation coefficients were negative. Conclusion These results suggest that people who reside in Shanghai are exposed to phthalates, especially to DBP and DEHP. Although the level of phthalates is relatively mild, an association of phthalate levels and reduced quality of human semen has been shown in the present study.