Formulation and evaluation of cyclodextrin-based drug delivery system for oral application of zaleplon

Nesanica je jedan od najčešćih poremećaja spavanja koji zahvaća oko 30% odrasle populacije, a značajno je češći kod starije populacije i psihičkih bolesnika. Zahvaljujući svojim farmakološkim svojstvima, zaleplon je hipnotik izbora za kratkotrajno liječenje nesanice. Međutim, slaba topljivost i razgradnja u jetri ograničavaju mu oralnu bioraspoloživost na samo 30%. Stoga je cilj ovog rada pripremom inkluzijskih kompleksa s ciklodekstrinima poboljšati topljivost lijeka u vodi, a uklapanjem u oblik s kontroliranim oslobađanjem razviti novi terapijski sustav koji bi mogao značajno unaprijediti terapijsku učinkovitost ovog lijeka. Spektrofluorimetrijska i solubilizacijska ispitivanja su pokazala da β-ciklodekstrin sa zaleplonom stvara komplekse prikladne stabilnosti, no topljivost nastalog kompleksa je ograničena. Od kemijski modificiranih derivata, nasumično metilirani β-ciklodekstrin se pokazao najprikladnijim, dok je topljivost i stabilnost inkluzijskih kompleksa zaleplona sa sulfobutil-β-ciklodekstrinom i hidroksipropil-β-ciklodekstrinom nešto niža. Nastajanje inkuzijskih kompleksa s navedenim derivatima ciklodekstrina je potvrđeno primjenom 1H NMR spektroskopije, a određena je i njihova struktura. Sustav karakterizira istovremeno prisustvo dva načina vezanja. Prvi, dominantni način uključuje inkluziju fenilnog prstena molekule zaleplona u centralnu šupljinu molekule ciklodekstrina, a drugi inkluziju pirazolo[1,5-a] pirimidinskog prstena molekule lijeka, dok je nastajanje kompleksa višeg reda isključeno. Termodinamička ispitivanja su pokazala da je nastajanje kompleksa zaleplona s β-ciklodekstrinom i njegovim metiliranim i hidroksipropiliranim derivatima entalpijom uvjetovani proces, dok je kompleksiranje lijeka sa sulfobutil- β-ciklodekstrinom entropijom vođen proces. U oba slučaja riječ je o spontanom procesu. Dodatkom hipromeloze je značajno povećan solubilizacijski potencijal metiliranog β-ciklodekstrina uslijed nastajanja ternarnih kompleksa lijek-ciklodekstrin-polimer, dok hidrofilni polimeri nisu utjecali na topljivost i stabilnost kompleksa zaleplona s β-ciklodekstrinom. Binarni i ternarni kompleksi zaleplona s navedenim ciklodekstrinima i hidrofilnim polimerima u čvrstom stanju pripremljeni su tehnikom sušenja raspršivanjem. Diferencijalna pretražna kalorimetrija, difrakcija rentgenskih zraka na prašku te pretražna elektronska mikroskopija su pokazale djelomičnu amorfizaciju lijeka u binarnim i ternarnim kompleksima s β-ciklodekstrinom, dok su kompleksi s metiliranim-β-ciklodekstrinom bili u potpunosti amorfne strukture. Binarni kompleksi s metiliranim-β- ciklodekstrinom te ternarni kompleksi s metiliranim-β-ciklodekstrinom i hipromelozom bili su najučinkovitiji u povećanju topljivosti zaleplona u odnosu na ostale komplekse. Korištenjem manitola kao topljivog punila te ternarnog kompleksa zaleplona s metiliranim β-ciklodekstrinom i hipromelozom pripremljene su tablete koje omogućuju oslobađanje doze lijeka u samo 5 minuta, osiguravajući brz početak djelovanja lijeka. Topljivi kompleksi zaleplona s u vodi topljivim, polimernim derivatom ciklodekstrina su pripremljeni i tehnikom mljevenja u vibracijskim mikromlinovima. Nakon 90 minuta mljevenja pri frekvenciji od 24 Hz dobiven je amorfni produkt koji otapanjem u vodi prelazi u inkluzijski kompleks, što je dokazano 1H-NMR spektroskopijom. Primjenom tehnike sušenja raspršivanjem pripremljene su mikrosfere s pH ovisnim oslobađanjem zaleplona kao novi terapijski sustav za liječenje nesanice koju karakterizira prerano buđenje te nemogućnost naknadnog usnivanja. Matriks koji sadrži 80% Eudragita S100 i 20% glicerolmonostearata pokazao se optimalnim u sprječavanju neželjenog oslobađanja lijeka pri nižim pH-vrijednostima medija, dok je uklapanjem 50% doze lijeka u obliku inkluzijskog kompleksa s metiliranim β-ciklodekstrinom osigurano potpuno oslobađanje lijeka u umjetnom crijevnom mediju, kinetikom nultog reda (k0=155 μg/min).Insomnia is one of the most common sleep disorders affecting about 30% of adult population, while its prevalence is even higher among elderly and psychiatric patients. Pharmacological properties makes zaleplon the drug of choice for the short term treatment of insomnia, but its low aqueous solubility and intensive metabolism in liver are reducing its oral bioavailability to only 30%. Therefore, the aim of this work was to improve aqueous solubility of zaleplon through inclusion complexation and to develop a novel drug delivery system able to enhance its therapeutical efficiency. Spectrofluorimetric and phase solubility studies showed that parent β-cyclodextrin forms stable inclusion complexes with zaleplon, but aqueous solubility of the complexes formed was limited. Among chemically modified β- cyclodextrin derivatives, randomly methylated derivative was the most efficient, while the solubility and stability of zaleplon complexes with sulphobutyl- and hydroxypropyl-β-cyclodextrin was less pronounced. The inclusion complex formation in all cases was confirmed by 1H-NMR spectroscopy. The structure of the complexes formed was characterised by the presence of two different binding modes which existed simultaneously in the solution. The first and the dominant one occurs through inclusion of phenyl moiety of the drug into central cavity of cyclodextrin molecule, while the other involves the inclusion of the pyrazolo[1,5-a]pyrimidine ring of the drug. The formation of higher order complexes was not demonstrated. Thermodynamic studies showed that complexation of zaleplon with β- cyclodextrin and its randomly methylated and hydroxypropylated derivatives was enthalpy driven, while entropy driven complexation was observed in case of sulphobutyl-β-cyclodextrin. In all cases, inclusion complex formation was a spontaneous process. Addition of hypromellose enhanced significantly the solubilising and complexing potential of randomly methylated β-cyclodextrin due to ternary complex formation, while hydrophilic polymers had no effect on solubility and stability of zaleplon complexes with parent β-cyclodextrin. Binary and ternary zaleplon complexes with cyclodextrins and polymers tested were prepared by spray-drying. Differential scanning calorimetry, X-ray powder diffractometry and scanning electron microscopy showed only partial drug amorphization of the drug in binary and ternary complexes with β-cyclodextrin, while all complexes with randomly methylated β-cyclodextrin were completely amorphous. The inclusion complex formation in spray-dried products was confirmed by the solid state NMR. Binary and ternary complexes with randomly methylated β-cyclodextrin and hypromellose were the most efficient in increasing the dissolution rate of zaleplon. The use of mannitol as a soluble filler and ternary inclusion complex with randomly methylated β-cyclodextrin and hypromellose allowed formulation of tablets which released the complete drug dose in 5 minutes, providing fast onset of drug action. Highly soluble zaleplon complexes with soluble, polymeric β-cyclodextrin derivative were also prepared by co-grinding in high energy vibrational micromill. Co-grinding at frequency of 24 Hz resulted in an amorphous product which is readily soluble in water, forming inclusion complexes as demonstrated by 1H-NMR spectroscopy. Spray-drying technique was used to prepare pH-responsive zaleplon microspheres with delayed release properties as a novel drug delivery system for treatment of specific type of insomnia characterised with premature awaking and inability to fall asleep again. Matrix containing 80% of Eudragit S100 and 20% of glycerol monostearate was demonstrated to be optimal, preventing the drug release at high pH-values. The incorporation of 50% of the drug dose in form of binary inclusion complex with randomly methylated β-cyclodextrin ensured complete drug release in simulated intestinal media, with the zero-order release rate (k0=155 μg/min)

Оblici komunikacije slepogluvih osoba

Slepogluvoća je kombinacija oštećenja vida i oštećenja sluha koja dovodi do teškoća u domenima komunikacije, mobilnosti i svakodnevnog funkcionisanja. Potpuno ili delimično ograničava osobu u aktivnostima i učešću u tolikoj meri da društvo treba da joj obezbedi posebne usluge, izmene u okruženju i asistivne tehnologije. Slepogluve osobe pripadaju izuzetno heterogenoj grupi osoba sa invaliditetom. Deca, mladi i odrasli se razlikuju prema vrsti, stepenu i vremenu nastanka oštećenja vida i oštećenja sluha, fizičkim i zdravstvenim problemima, kognitivnom funkcionisanju, oblicima komunikacije koje koriste, postojanju dodatnih smetnji u razvoju. Pristupi i intervencije u obrazovanju i rehabilitaciji slepogluvih osoba su proistekli iz područja koja čine obrazovanje i rehabilitacija osoba sa oštećenjem vida i obrazovanje i rehabilitacija osoba sa oštećenjem sluha. Međutim, slepogluve osobe zahtevaju podršku i sistem usluga koje se značajno razlikuju od onih namenjenih isključivo osobama sa oštećenjem vida ili osobama sa oštećenjem sluha. U cilju razvijanja i održavanja komunikacije slepogluvih osoba kreiran je širok spektar podrške, alternativnih komunikacionih sistema i tehnika i razvijen znatan broj oblika komunikacije. Oblici komunikacije koje slepogluve osobe mogu da koriste su: govor, znakovni jezik, locirani znakovni jezik, vođeni znakovni jezik, prstna azbuka/abeceda, ručna azbuka/abeceda, taktilni znakovni jezik, Lormova abeceda, pisanje na dlanu, Tadoma metod, pisani govor, uvećana štampa, Brajevo pismo. Cilj rada je opisati oblike komunikacije koje mogu da razviju i koriste slepogluve osobe

The development of inclusive education with regard to teacher competencies: Current situation, problems and perspectives

This paper aims to point out the importance of acquiring and developing teacher competencies for work with children with disabilities in an inclusive educational environment. After providing an overview of educational inclusion as the goal and content of current reform processes both in Serbia and around the world, the paper examines this process from the standpoint of teachers and their inclusion competencies. An insight into different studies and research results on teachers' roles, attitudes and competencies indicates that educational inclusion in our country has not taken hold to a sufficient extent, and that teachers express serious doubts regarding their competencies to work with children from sensitive social groups. In practice, teachers, as well as students of teacher education faculties are not sufficiently familiar with the fundamental principles of the inclusive approach to education, and emphasize that they are more ready to work with children with mild disabilities. On the other hand, they are of the opinion that children with multiple disabilities should be educated in specialized institutions. In conclusion, the development of the concept and practice of inclusive education will require serious interventions, both in terms of initial teacher education, and in terms of promoting their role in the contemporary education system for children with disabilities.Cilj rada je da se ukaže na značaj razvoja i sticanja nastavničkih kompetencija za rad sa decom sa smetnjama i teškoćama u razvoju u inkluzivnom obrazovnom okruženju. U radu se, nakon osvrta na inkluziju u obrazovanju kao cilju i sadržaju aktuelnih reformskih procesa kod nas i u svetu, taj proces razmatra iz ugla nastavnika i njegovih kompetencija za inkluziju. Uvid u studije i rezultate istraživanja o ulogama, stavovima i kompetencijama nastavnika ukazuje da proces inkluzije u našoj zemlji još nije dovoljno zaživeo i da nastavnici ispoljavaju ozbiljne sumnje u svoje kompetencije za rad sa decom iz osetljivih društvenih grupa. Nastavnici u praksi, kao i studenti nastavničkih fakulteta, nemaju jasan doživljaj suštine inkluzivnog pristupa u obrazovanju i ističu da su spremniji za rad sa decom sa lakšim oblicima smetnji, dok obrazovanje dece sa višestrukim smetnjama treba da se odvija u specijalizovanim ustanovama. U cilju razvoja ideje i prakse inkluzivnog obrazovanja, neophodno je uraditi ozbiljne intervencije kako na planu inicijalnog obrazovanja nastavnika tako i na planu promovisanja njihove uloge u savremenom sistemu obrazovanja dece sa smetnjama i teškoćama u razvoju

Thermal analysis of N-carbamoyl benzotriazole derivatives

Thermal properties of N-carbamoyl benzotriazole derivatives and N,N',N''-tribenzyloxyisocyanuric acid were investigated using thermogravimetric analysis and differential scanning calorimetry. The results revealed a difference between structural analogs of N-carbamoyl benzotriazole derivatives. They seem to be in agreement with the previously proposed formation of N,N',N''-tribenzyloxyisocyanuric acid from 1-(N-benzyloxycarbamoyl) benzotriazole, via an intermediary N-benzyloxyisocyanate acid, during heating. Substantially different thermal properties were observed for structural analogues, 1-(N-methoxycarbamoyl) benzotriazole and 1-(N-ethoxycarbamoyl) benzotriazole. In contrast to N-benzyloxyisocyanate, no corresponding reactions were observed for their decomposition products, i.e., methoxyisocyanate and ethoxyisocyanate

Comparison of Zn Content in Rapid-acting Insulin and Biphasic Suspension by FAAS

Various insulin analogs (rapid-acting and intermediate-acting insulin) have been investigated for the determination of zinc content by flame atomic absorption spectrometry. This paper presents the validation of a method and comparison of zinc content in the insulin samples studied. The method was linear (r)(^2) = 0.9997), the limit of detection was 0.0098 mg L(^{–1}) and the limit of quantification was 0.0296 mg L(^{–1}), the precision (as relative standard deviation) was up to 7.4 %, and the accuracy was within a range of 95.6 % to 100.1 % for the recovery of fortified insulin samples. The zinc content in the insulin samples ranged from 14.9 mg L(^{–1}) to 16.3 mg L(^{–1}) for rapid-acting insulin and 18.7 mg L(^{–1}) to 19.9 mg L(^{–1}) for intermediate-acting insulin. A higher zinc content was found in the intermediate-acting insulins than in the rapid-acting insulin analogs ((p) < 0.05). The obtained results should be considered in the establishment of new or improvement of currently available procedures used to assure the quality, safety and efficacy of insulin products. This work is licensed under a Creative Commons Attribution 4.0 International License

Podsticanje razvoja koordinacije kod učenika sa smetnjama u razvoju u redovnoj osnovnoj školi

The paper deals with the effects of specially programmed types of physical activities on the development of coordination of the pupils with developmental difficulties in regular elementary schools. The method of pedagogical research with parallel groups was used in the research which included 135 junior elementary-school pupils. Among them 90 pupils had one of developmental difficulties (intellectual disability, visual and hearing disturbances) and they were divided into an experimental and a control group 1, while other pupils had typical development and they formed the control group 2 (n=45). Besides regular classes of physical education the experimental group was engaged in a special programme, devised and implemented into the Individual Educational Plan, as one of the types of intervention within the system of inclusive education. The research lasted for three months during which the groups were tested by the tests of coordination and the data obtained at the beginning and the end of the research period were processed by the application of univariant analysis of covariance and the T-test of the paired samples. The results show that the pupils with developmental difficulties can considerably progress in the development of coordination if their individual characteristics are recognized and optimal conditions are provided.U ovom radu ispituje se uticaj primene posebno programiranih oblika fizičke aktivnosti na razvoj sposobnosti koordinacije kod učenika sa smetnjama u razvoju u redovnoj osnovnoj školi. Istraživanje je sprovedeno metodom pedagoškog eksperimenta sa paralelnim grupama na uzorku od 135 učenika mlađeg školskog uzrasta. Od toga, kod 90 učenika je evidentirana jedna od smetnji u razvoju (intelektualna smetnja, vizuelna smetnja, smetnja sluha) i oni su podeljeni u eksperimentalnu i kontrolnu grupu 1, dok su ostali učenici imali tipičan razvoj i činili su kontrolnu grupu 2 (n = 45). Eksperimentalna grupa je pored časova redovne nastave fizičkog vaspitanja bila uključena u poseban program vežbanja koji je osmišljen i postavljen u okviru Individualnog obrazovnog plana (IOP) kao jednog od vidova intervencije predviđenim inkluzivnim sistemom obrazovanja. Istraživanje je trajalo tri meseca, u okviru koga su grupe testirane testovima koordinacije, a podaci dobijeni na početku i kraju istraživanja obrađeni su primenom univarijantne analize kovarijanse i T-testa uparenih uzoraka. Rezultati ukazuju da učenici sa smetnjama u razvoju mogu znatno da napreduju u razvoju sposobnosti koordinacije pod uslovom da se uvaže njihove individualne karakteristike i obezbede optimalni uslovi

Functional approach and learning mathematics for children with multiple disabilities

U širem smislu, matematika predstavlja nauku o količini, strukturi, prostoru i promenama. Kao nastavni predmet izučava se na različitim nivoima školovanja. Usvajanje matematičkih znanja je važno za obrazovanje, ali i za svakodnevni život, što objašnjava značaj koji ovaj predmet zauzima u nastavnom planu i programu. Kada se govori o tome koji su ciljevi i ishodi nastave matematike sa učenicima sa višestrukim smetnjama, važno je istaći da oni prate funkcionalni kurikulum u okviru kojeg se učenje zasniva upravo na aktivnostima koje se realizuju u prirodnom okruženju. Učenjem kroz uključivanje u realne, svakodnevne situacije podstiču se samostalnost i socijalna participacija ovih učenika. Isticanje načina i mesta na kojima se nastava odvija je specifičnost funkcionalng kurikuluma. Cilj rada je prikazati pristupe i strategije za usvajanje matematičkih sadržaja učenika sa višestrukim smetnjama. Podaci su prikupljeni pregledom dostupnih štampanih izvora i elektronskih baza ERIC, JSTOR, Google Scholar, sa odgovarajućim ključnim rečima. Heterogenost populacije višestruko ometenih ne omogućava primenu univerzalnih metoda rada, već isključivo individualni pristup radi sticanja znanja. Fokus treba da bude na snagama i interesima učenika, a ide se ka oblastima u kojima je potrebno poboljšanje, bez obzira na hronološki uzrast. Učenje treba da se odvija kroz prirodne rutine, u svakodnevnim situacijama, kroz socijalne interakcije, uz primenu određenih strategija.In a broader sense, mathematics is the science of quantity, structure, space, and change. As a school subject, it is studied at different levels of education. Acquisition of mathematical knowledge is important for education, but also for everyday life, which explains the importance of this subject in the curriculum. When it comes to goals and outcomes of teaching mathematics to students with multiple disabilities, it is important to point out that they follow a functional curriculum in which learning is based on activities that are realized in the natural environment. Learning through involment in real, everyday activities encourages the independence and social participation of these students. Emphasizing the ways and places where teaching takes place is a specific feature of the functional curriculum. The aim of this paper is to present approaches and strategies for the adoption of mathematical content of students with multiple disabilities. Data was collected by reviewing available printed sources and electronic databases ERIC, JSTOR, Google Scholar, with appropriate keywords. The heterogeneity of the population with multiple disabilities does not allow the application of universal methods of work, instead, an individual approach should be used for knowledge acquisition. The focus should be on students’ strengths and interests, and the goal is to master the areas where improvement is needed, regardless of chronological age. Learning should take place through natural routines, in everyday situations, through social interactions, with the application of certain strategie

Functional approach and learning mathematics for children with multiple disabilities

U širem smislu, matematika predstavlja nauku o količini, strukturi, prostoru i promenama. Kao nastavni predmet izučava se na različitim nivoima školovanja. Usvajanje matematičkih znanja je važno za obrazovanje, ali i za svakodnevni život, što objašnjava značaj koji ovaj predmet zauzima u nastavnom planu i programu. Kada se govori o tome koji su ciljevi i ishodi nastave matematike sa učenicima sa višestrukim smetnjama, važno je istaći da oni prate funkcionalni kurikulum u okviru kojeg se učenje zasniva upravo na aktivnostima koje se realizuju u prirodnom okruženju. Učenjem kroz uključivanje u realne, svakodnevne situacije podstiču se samostalnost i socijalna participacija ovih učenika. Isticanje načina i mesta na kojima se nastava odvija je specifičnost funkcionalng kurikuluma. Cilj rada je prikazati pristupe i strategije za usvajanje matematičkih sadržaja učenika sa višestrukim smetnjama. Podaci su prikupljeni pregledom dostupnih štampanih izvora i elektronskih baza ERIC, JSTOR, Google Scholar, sa odgovarajućim ključnim rečima. Heterogenost populacije višestruko ometenih ne omogućava primenu univerzalnih metoda rada, već isključivo individualni pristup radi sticanja znanja. Fokus treba da bude na snagama i interesima učenika, a ide se ka oblastima u kojima je potrebno poboljšanje, bez obzira na hronološki uzrast. Učenje treba da se odvija kroz prirodne rutine, u svakodnevnim situacijama, kroz socijalne interakcije, uz primenu određenih strategija.In a broader sense, mathematics is the science of quantity, structure, space, and change. As a school subject, it is studied at different levels of education. Acquisition of mathematical knowledge is important for education, but also for everyday life, which explains the importance of this subject in the curriculum. When it comes to goals and outcomes of teaching mathematics to students with multiple disabilities, it is important to point out that they follow a functional curriculum in which learning is based on activities that are realized in the natural environment. Learning through involment in real, everyday activities encourages the independence and social participation of these students. Emphasizing the ways and places where teaching takes place is a specific feature of the functional curriculum. The aim of this paper is to present approaches and strategies for the adoption of mathematical content of students with multiple disabilities. Data was collected by reviewing available printed sources and electronic databases ERIC, JSTOR, Google Scholar, with appropriate keywords. The heterogeneity of the population with multiple disabilities does not allow the application of universal methods of work, instead, an individual approach should be used for knowledge acquisition. The focus should be on students’ strengths and interests, and the goal is to master the areas where improvement is needed, regardless of chronological age. Learning should take place through natural routines, in everyday situations, through social interactions, with the application of certain strategie

Inclusion in physical education and sport

Postoji međunarodna saglasnost za ideju da fizička aktivnost i sport mogu predstavljati strategije društvene inkluzije, kao i da osobe sa smetnjama i teškoćama u razvoju poseduju kapacitete za uče-stvovanje u ovim aktivnostima. Ipak, procesi kroz koje intervencije zasnovane na fizičkoj aktivnosti i sportu mogu uticati na inkluzivne ishode još uvek zahtevaju sveobuhvatnu i kontinuiranu istragu. Cilj ovog rada je da kroz pregled izvora naučne produkcije sa međunarodnih naučnih skupova, naučnih časopisa iz oblasti sportskih i društvenih nauka, kao i referentne literature, ukaže na probleme i perspektive razvoja nastave inkluzivnog fizičkog vaspitanja i sporta. Podaci su prikupljeni pregle-dom dostupnih štampanih izvora i elektronskih baza ERIC, JSTOR, Google Scholar, sa odgovarajućim ključnim rečima. Rezultati analize ukazuju da postoji više faktora koji utiču na nastavu fizičkog vaspitanja u inklu-zivnom obrazovnom kontekstu (stavovi nastavnika i učenika, kompetencije nastavnika, pristupačnost i sl.) kao i da je inkluzija u sportu uslovljena kako ličnim (priroda same smetnje, zdravlje, motivacija i sl.) tako i fak-torima vezanim za društvenu sredinu (pristupačnost, kreiranje i primena adaptiranih fizičkih aktivnosti i sl.).There is an international consensus that physical activity and sport can be used as strategies for social inclusion, as well as that people with disabilities have the capacity to participate in these activities. However, interventions based on physical activity and sport and their processes that can influence inclusive outcomes are yet to be continuously and comprehensively investigated. The aim of this paper is to point out the problems and perspectives of the development of inclusive physical education and sport by providing an overview of scientific production sources from international scientific conferences, scientific journals in the field of sports and social sciences, as well as reference literature. Data were collected by reviewing available print sources and electronic databases suh asERIC, JSTOR, and Google Scholar etc., using relevant keywords. The results indicate that there are several factors which influence physical education and its teaching methods in an inclusive educational context (attitudes of students and teachers, teachers’ competencies, approachability, etc.), and that inclusion in sport is dependent on both personal (nature of the disability, health, motivation, etc.) and social environment factors (accessibility, creating and implementing planned physical activities adapted for students with disabilities, etc.)

Spectrophotometric Determination of Malondialdehyde in Urine Suitable for Epidemiological Studies

A reliable method for spectrophotometric determination of urinary malondialdehyde (MDA), according to the thiobarbituric acid (TBA) assay, is described. To account for matrix interference and differences in individual urine composition, standard addition procedure was applied. The method is adequately selective (LoQ = 0.09 μM in the presence of 0.1 M creatinine and 0.5 M urea) and reliable (within-day and between-day variability of less than 5 %). The mean level of urinary MDA was 1.52 ± 0.73 µM that is in good agreement with spectrofluorometric determination (1.20 ± 0.56 μM; p = 0.085) as well as with previous studies that used HPLC. Furthermore, it is demonstrated that MDA is stabile in urine at room temperature for 24 h and when stored at –20 °C for 6 months. The described method enables simple, rapid and cost-effective determination of urinary MDA as a relevant and non-invasive marker of “whole-body” oxidative stress. This work is licensed under a Creative Commons Attribution 4.0 International License