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Quality control of Verbasci flos
No full textU okviru ovog diplomskog rada uspoređena su dva uzorka droge Verbasci flos sakupljena na dvije različite lokacije te je provedena kontrola kakvoće tih uzoraka. Određivanjem broj bubrenja i gubitka sušenjem ustanovljeno je da uzorci odgovaraju farmakopejskim zahtjevima, dok je u uzorku 2 pronađena nešto veća količina fragmenta čaške i ostalih stranih tvari. Mikroskopskim pregledom pronađeni su svi dijelovi koje Farmakopeja propisuje. Tankoslojnom kromatografijom (TLC) dokazana je prisutnost flavonoida i fenolnih kiselina koje su potom određene UV-Vis spektrofotometrijskom metodom te je sadržaj iznosio 0,053-0,075% i 0,08- 0,24%. Također određen je i sadržaj trjeslovina koji je iznosio 0,172-0,24%.As part of this thesis, two types of mullein flower from two different locations were compared and the quality control of these samples was carried out. By determining the swelling index and loss on drying, it was found that the samples correspond to pharmacopoeial requirements, while in the sample 2 there was slightly larger amount of fragments of the calyx, and other foreign substances was found. By microscopic examination we found all the parts prescribed by the Pharmacopoeia. Thin-layer chromatography (TLC) showed the presence of flavonoids and phenolic acids, which were then quantified by UV-vis spectrometric method, with the content ranging from 0.053% to 0.075% and 0.08% to 0.24%. The content of tannins was also determined, ranging from 0.172% to 0.24%
Validation of MEKC-MS/MS method for determination of abemaciclib, palbociclib and ribociclib in blood plasma
No full textHR+/HER2- rak dojke jedan je od najčešćih tipova karcinoma u svijetu, za čije se liječenje koristi kombinacija inhibitora CDK4/6 s endokrinom terapijom. Abemaciklib, palbociklib i ribociklib, selektivni su i reverzibilni inhibitori o ciklinima ovisnih kinaza, CDK 4 i 6.
S obzirom na potrebu za istovremenim određivanjem ovih lijekova iz krvne plazme pacijentica, razvijena je metoda micelarne elektrokinetičke kromatografije spregnute s tandemskom spektrometrijom masa (MEKC-MS/MS). Kako bi se navedena metoda mogla implementirati u rutinske analize biokemijskih laboratorija i sa sobom donijeti prednosti visoke efektivnosti, selektivnosti i manjeg utroška materijala te omogućiti terapijsko praćenje inhibitora CDK4/6 s ciljem poboljšanja ishoda liječenja, bilo ju je potrebno validirati sukladno ICH M10 smjernicama za validaciju bioanalitičkih metoda.
Linearnost, točnost i preciznost metode su validirane za abemaciklib u kalibracijskom području 50-500 ng/mL, za palbociklib 30-300 ng/mL i za ribociklib 500-5000 ng/mL. Na visokoj i niskoj QC koncentraciji, validirani su parametri utjecaja matrice i stabilnost uzoraka u autosampleru za svaki od tri inhibitora CDK4/6. Zaključno s validacijom ovih parametara, pokazano je kako je metoda prikladna za svoju namjenu.HR+/HER- breast cancer is one of the most common types of carcinomas across the world, for which the therapy includes combination of CDK4/6 inhibitors with endocrine therapy. Abemaciclib, palbociclib and ribociclib are selective and reversible inhibitors of cyclin-dependent kinases, CDK 4 and 6.
Due to the need for simultaneous determination of these drugs from the blood plasma of patients, a method of micellar electrokinetic chromatography coupled with tandem mass spectrometry (MEKC-MS/MS) was developed. In order for the above method to be implemented in routine analyses in biochemical laboratories and to bring with it the advantages of higher effectiveness, selectivity and lower material consumption, while enabling therapeutic monitoring of CDK4/6 inhibitors with the aim of improving treatment outcomes, it was necessary to validate it in accordance with ICH M10 Guideline for the validation of bioanalytical methods.
Linearity, accuracy, and precision of the method were validated in the calibration range of 50-500 ng/mL for abemaciclib, 30-300 ng/mL for palbociclib and 500-5000 ng/mL for ribociclib. Validation parameters matrix effect and stability in autosampler were validated for all of the three CDK4/6 inhibitors on the high and low QC (Quality control) concentrations, respectively to the ICH M10 Guideline. In conclusion, with validation of these parameters, it was shown that the method is suitable for its purpose
Determination of potassium and sodium in dietary supplements therapies for inflammatory bowel diseases by atomic absorption spectrometry
No full textDodaci prehrani u širokoj su primjeni i važni su za održavanje zdravlja kod ljudi. Budući da više od
60 % bolesnika koristi dodatke prehrani u ublažavanju simptoma upalnih bolesti crijeva, cilj ovog
rada bio je procijeniti kvalitetu dodataka prehrani s obzirom na sadržaj kalija i natrija. Sadržaj
navedenih makroelemenat u dodacima prehrani koji sadrže biljne vrste: indijski tamjanovac,
kurkumu, justiciju i crni papar određen je primjenom plamene tehnike atomske apsorpcijske
spektrometrije nakon provedene mikrovalne digestije. Rezultati istraživanja pokazali su dominantnu
zastupljenost kalija i značajno manji sadržaj natrija te širok raspon oba makroelementa u
analiziranim dodacima prehrani. Rezultati su pokazali manji sadržaja kalija u suhim ekstraktima
nego u usitnjenim biljnim drogama. Dodaci prehrani za liječenje upalnih bolesti crijeva mogu se
smatrati dobrim izvorom minerala, a pozitivan omjer Na/K primjenu i kod drugih bolesnika.Dietary supplements are widely used and are important for maintaining people's health. Since more
than 60% of patients use dietary supplements to alleviate the symptoms of inflammatory bowel
diseases, this work aimed to evaluate the quality of dietary supplements with regard to potassium
and sodium content. The content of the mentioned macroelements in dietary supplements containing
botanical species, Indian frankincense, turmeric, black paper, and justice was determined using the
flame technique of atomic absorption spectrometry after microwave digestion. The obtained results
showed a dominant presence of potassium, a significantly lower sodium content, and a wide range
of macroelements in the analyzed dietary supplements. The results showed a lower potassium
content in dry extracts than in crushed herbal drugs. Dietary supplement therapies for inflammatory
bowel diseases can be considered a good source of minerals, and a positive Na/K ratio can also be
used in other patient
Phytochemical composition and antioxidant activity of fruit of Ribes uva-crispa L. (Grossulariaceae)
No full textOvim diplomskim radom analiziran je fitokemijski sastav te je ispitano antioksidacijsko djelovanje plodova vrste Ribes uva-crispa L. Identifikacija flavonoida (rutina, kvercitrina i izokvercitrina) i fenolnih kiselina (klorogenske kiseline) provedena je primjenom tankoslojne kromatografije. U vodenom i metanolnom ekstraktu plodova R. uva-crispa L. utvrđena je prisutnost rutina i izokvercitrina te klorogenske kiseline. Kvantifikacija fenolnih kiselina (izraženih kao klorogenska kiselina) i ukupnih polifenola (izraženih kao galna kiselina) provedena je spektrofotometrijskim metodama.
Sadržaj fenolnih kiselina, izražen kao klorogenska kiselina iznosio je 0,45 %, dok je udio ukupnih polifenola, izražen kao galna kiselina iznosio 2,5 %. Antioksidacijsko djelovanje ekstrakta ispitano je dvjema spektrofotometrijskim metodama: određivanjem sposobnosti hvatanja DPPH slobodnog radikala (IC50: 331,56 ± 15,90 μg/mL,) te određivanjem sposobnosti hvatanja NO radikala (IC50: 332,22 ± 11,22 μg/mL).
Ovaj rad potvrdio je fitoterapijski potencijal vrste Ribes uva-crispa L.In this master's thesis the phytochemical composition and antioxidant activity of the fruits of Ribes uva-crispa L. were analyzed. The identification of flavonoids (rutin, quercitrin and isoquercitrin) and phenolic acids (chlorogenic acid) was conducted using thin-layer chromatography. The presence of rutin, isoquercitrin and chlorogenic acid was determined in the water and methanol extracts of R. uva-crispa L. fruits. The quantification of phenolic acids (expressed as chlorogenic acid) and total polyphenols (expressed as gallic acid) was performed using spectrophotometric methods. The content of phenolic acids, expressed as chlorogenic acid was 0,45 %, while the amount of total polyphenols, expressed as gallic acid was 2,5 %. The antioxidant activity of extracts was defined using two spectrophotometric methods: the determination of the DPPH free radical scavenging ability (IC50: 331,56 ± 15,90 μg/mL) and the determination of the NO radical scavenging ability (IC50: 332,22 ± 11,22 μg/mL).
This study confirmed the phytotherapeutic potential of Ribes uva-crispa L
Determination of metals in dietary supplements used for the treatment of inflammatory bowel diseases using graphite furnace technique
No full textPojavnost upalnih bolesti crijeva je sve učestalija u posljednjih nekoliko desetljeća širom svijeta, a
samim time raste i potražnja i potrošnja dodataka prehrani koji imaju blagotvorni učinak na
liječenje bolesti. Bilo da se koriste kao suportivna terapija ili primarna terapija iznimno je važno da
su takvi pripravci dobro kontrolirani i da je poznat njihov kompletan sastav kako bi se osigurala
sigurnost korisnika. Metali se dijele na esencijalne i neesencijalne. Esencijalni su neophodni za
život ljudi u dobro definiranim količinama. Svi metali su toksični u većim dozama i mogu izazvati
kronične metaboličke poremećaje u organizmu. Upravo iz tog razloga nastoji se što bolje
kontrolirati njihov sadržaj u lijekovima, dodacima prehrani i hrani. Cilj rada bio je analizirati
dodatke prehrani koji se koriste za liječenje upalnih bolesti crijeva. Analizirani dodaci prehrani
sadrže crni papar, indijski tamjanovac, justiciju i/ili kurkumu kao biljne droge, a prikupljeni su u
trgovinama zdravom hranom ili putem internet trgovine. Za određivanje sadržaja kroma,
molibdena i kositra korištena je grafitna atomska apsorpcijska spektrometrija nakon uklanjanja
složene matrice spaljivanjem uzoraka mikrovalnom digestijom. Sadržaj metala u svim
analiziranim uzorcima bio je ispod propisanih maksimalnih vrijednosti propisanim od strane
regulatornih tijela.The prevalence of inflammatory bowel diseases has been increasing globally over the past few
decades, leading to a growing demand and consumption of dietary supplements with beneficial
effects on disease treatment. Whether used as supportive or primary therapy, it is crucial that such
preparations are well-regulated and their complete composition is known to ensure user safety.
Metals are categorized into essential and non-essential. Essential metals are necessary for human
life in well-defined quantities. All metals are toxic in larger doses and can cause chronic metabolic
disorders in the body. Therefore, the content of drugs, dietary supplements, and food is strictly
controlled. The aim of this study was to analyze dietary supplements used for treating
inflammatory bowel diseases. The analyzed supplements contain black pepper, Indian
frankincense, justice, and/or turmeric as herbal drugs, and were collected from health food stores
or through online shops. Graphite atomic absorption spectrometry was used after removing the
complex matrix by microwave digestion of the samples to determine the content of chromium,
molybdenum, and tin. The metal content in all analyzed samples was below the maximum values
prescribed by regulatory bodies
Biological activities of the plant species Globularia alypum L.
No full textMeđu vrstama roda Globularia L. najviše istražena je vrsta Globularia alypum L. zbog svoje široke primjene u tradicionalnoj medicini za liječenje mnogih bolesti. S obzirom na sve veći broj istraživanja o ovoj biljnoj vrsti, u ovom diplomskom radu okupljeni su svi originalni znanstveni radovi objavljeni u periodu od 2017. do 2023. godine koji su ispitivali biološka djelovanja njezinih ekstrakata. Pretraživanjem 12 baza podataka ukupno je pronađeno 38 radova. Među svim zabilježenim djelovanjima (77), najčešće istraživana bila su antioksidacijsko (34 %), antibakterijsko (14 %) i protuupalno djelovanje (9 %). Brojni ekstrakti pokazali su značajno antioksidacijsko djelovanje (neki snažnije od standardnih antioksidansa). Metanolni ekstrakt pokazao je najširi spektar antibakterijskog djelovanja, dok je za različite ekstrakte zabilježeno antifungalno i antibiofilm djelovanje. Protuupalno djelovanje ekstrakata, popraćeno analgetskim djelovanjem, potvrđeno je inhibicijom proupalnih markera i citokina. Ekstrakti su također pokazali antipiretsko, citotoksično, antidijabetsko, antihiperlipidemijsko, nefroprotektivno, antihemolitičko, antikoagulacijsko, laksativno i antiartritičko djelovanje, a osim toga metanolni ekstrakt utjecao je na zacjeljivanje kože što ide u prilog njegovoj upotrebi u kozmetičkim pripravcima za topikalnu primjenu. Dobiveni rezultati ukazuju na to da biljna vrsta G. alypum može poslužiti kao izvor prirodnih antioksidansa, antibakterijskih i protuupalnih spojeva. S obzirom na zabilježena djelovanja njezinih ekstrakata, navedena vrsta pokazuje značajan terapijski potencijal, ali daljnja istraživanja potrebna su kako bi se bolje razumjeli mehanizmi djelovanja i optimalne doze ekstrakata.Among the species of the Globularia L. genus, Globularia alypum L. has been the most extensively studied due to its widespread use in traditional medicine for treating various diseases. Given the increasing number of studies on this plant, this diploma thesis compiles all original scientific articles that were published from 2017 to 2023 that have investigated biological activities of its extracts. Through searching 12 databases, a total of 38 studies were found. Among all documented activities (77), the most researched were antioxidant (34%), antibacterial (14%), and anti-inflammatory activity (9%). Numerous extracts showed significant antioxidant activity (some stronger than standard antioxidants). The methanolic extract exhibited the broadest spectrum of antibacterial activity, while various extracts showed antifungal and antibiofilm activities. The anti-inflammatory activity of extracts, accompanied by analgesic activity, was confirmed by the inhibition of inflammatory markers and cytokines. Extracts also demonstrated antipyretic, cytotoxic, antidiabetic, anti-hyperlipidemic, nephroprotective, anti-hemolytic, anticoagulant, laxative, and antiarthritic effects. Furthermore, the methanolic extract has been shown to promote skin healing, which goes in favor of its use in cosmetic products for topical application. The results obtained indicate that G. alypum plant species could serve as a source of natural antioxidants, antibacterial, and anti-inflammatory compounds. Considering the documented activities of its extracts, this species shows significant therapeutic potential, but further research is needed to better understand the mechanisms of action and optimal dosages of extracts
Development and validation of the HPLC-DAD-FLD method for the analysis of alpelisib in human plasma
No full textRak dojke predstavlja značajan javnozdravstveni problem zbog svojeg utjecaja na živote brojnih žena širom svijeta. U posljednjih
nekoliko godina, značajan napredak postignut je u liječenju raka dojke pozitivnog na hormonski receptor i negativnog na HER2 s
PIK3CA mutacijom korištenjem kombinacije alpelisiba s drugim lijekovima kao što su fulvestrant ili anastrozol. Alpelisib, inhibitor
PI3K-a, pokazuje velik potencijal u uspješnom liječenju ove vrste raka dojke, ali optimalni ishodi liječenja zahtijevaju učinkovito
praćenje terapije. U svrhu ovog rada razvijena je i validirana HPLC-DAD-FLD metoda za analizu alpelisiba u ljudskoj plazmi. Uzorci plazme pripremljeni su jednostavnom metodom taloženja proteina pomoću organskog otapala, što je osiguralo čistu analitnu matricu pogodnu za daljnju analizu. Korištena je kolona Phenomenex Gemini C18, dimenzija 150 × 4,6 mm i veličine čestica stacionarne faze 5 µm pri konstantnoj temperaturi od 25 °C. U razvijenoj metodi korištena je gradijentna eluacija mobilnom fazom koja se sastoji od 0,1 % mravlje kiseline u vodi i 0,1 % mravlje kiseline u metanolu. Brzina protoka iznosila je 1 mL/min. Korišteni su UV-Vis detektor i fluorescencijski detektor. Validacija metode provedena je ispitivanjem parametara linearnosti, točnosti, preciznosti i utjecaja matrice. Metoda je pokazala zadovoljavajuću linearnost unutar raspona koncentracija od 300 do 3000 ng/mL (r za DAD iznosi 0,99755, a za FLD 0,99664), visoku točnost (sve dobivene koncentracije čine od -1,8 do 11,3 % stvarne vrijednosti) i preciznost (RSD vrijednosti iznose od 1,1 do 4,3 %), te minimalan utjecaj matriksa (Meff se kreće od 95,8 do 106,1 %), čime je potvrđena njezina pouzdanost za analizu alpelisiba u kliničkim uzorcima. Konačno, razvijena i validirana metoda primijenjena je za analizu alpelisiba u plazmi tri pacijentice od kojih su rezultati za dvije pacijentice unutar validiranog raspona metode, a kod treće pacijentice alpelisib nije bilo moguće detektirati. Postoji mogućnost da pacijentica nije bila adherentna ili da je koncentracija u plazmi bila preniska da bi se ovom metodom mogla detektirati. Također je moguće da postoje interindividualne varijabilnosti koje utječu na koncentraciju alpelisiba u plazmi. Razvijena HPLC-DAD-FLD metoda za analizu alpelisiba u ljudskoj plazmi predstavlja napredak u mogućnostima praćenja terapije kod pacijentica s rakom dojke. Ova metoda može se koristiti za daljnja ispitivanja farmakokinetike alpelisiba, kao i za personalizaciju liječenja na temelju interindividualnih varijabilnosti među pacijenticama. Time se može postići bolja prilagodba terapije individualnim potrebama pacijentica, što će u konačnici rezultirati poboljšanjem ishoda liječenja i kvalitete života pacijentica oboljelih od raka dojke.Breast cancer represents a significant public health problem due to its impact on the lives of numerous women worldwide. In recent years, significant progress has been made in the treatment of hormone receptor-positive and HER2-negative breast cancer with PIK3CA mutation using a combination of alpelisib with other drugs such as fulvestrant or anastrozole. Alpelisib, a PI3K inhibitor, shows great potential in successfully treating this type of breast cancer, but optimal treatment outcomes require effective therapymonitoring. For the purpose of this study, an HPLC-DAD-FLD method was developed and validated for the analysis of alpelisib in human plasma. Plasma samples were prepared using a simple protein precipitation method with an organic solvent, ensuring a clean analytical matrix suitable for further analysis. A Phenomenex Gemini C18 column, dimensions 150 × 4.6 mm and particle size 5 µm, was used at a constant temperature of 25 °C. The developed method employed gradient elution with a mobile phase consisting of 0.1% formic acid in water and 0.1% formic acid in methanol. The flow rate was 1 mL/min. Both UV-Vis and fluorescence detectors were used. Method validation was carried out by examining parameters of linearity, accuracy, precision, and matrix effect. The method demonstrated satisfactory linearity within the concentration range of 300 to 3000 ng/mL (with an r value of 0.99755 for DAD
and 0.99664 for FLD), high accuracy (all obtained concentrations range from -1.8 to 11.3% of the true value) and precision (RSD values range from 1.1 to 4.3%), and minimal matrix effect (Meff ranges from 95.8 to 106.1%), confirming its reliability for the analysis of alpelisib in clinical samples.Finally, the developed and validated method was applied to analyze alpelisib in the plasma of three patients. The results for two patients were within the validated range of the method, while alpelisib could not be detected in the third patient's plasma. It is possible that the patient was not adherent to the therapy or that the plasma concentration was too low to be detected by this method. Additionally, inter-individual variability may affect the concentration of alpelisib in plasma.The developed HPLC-DAD-FLD method for analyzing alpelisib in human plasma represents an advancement in the capabilities of therapy monitoring for breast cancer patients. This method can be used for further pharmacokinetic studies of alpelisib and for the personalization of treatment based on inter-individual variability among patients. This approach can lead to better adjustment of therapy to the individual needs of patients, ultimately resulting in improved treatment outcomes and quality of life for breast cancer patients
Determination of non-essential elements in encapsulated dietary supplements for the treatment of inflammatory bowel diseases
No full textUpalne bolesti crijeva pogađaju sve veći broj pacijenata diljem svijeta. U današnje vrijeme brojni
pacijentu se okreću samoliječenju i posežu za brojnim dodacima prehrani. Provode se brojna
istraživanja kojima se dokazuje njihova korisnost. Međutim, jako je važno zaštiti pacijente i osigurati
kvalitetu dodataka prehrani propisanu smjernicama. Jedan od zahtjeva smjernica je i količina
elementarnih onečišćenja koja mogu imati toksične učinke na ljudsko zdravlje. Stoga je cilj ovoga
diplomskog rada bio odrediti sadržaj srebra, kositra i barija u dodacima prehrani za liječenje upalnih
bolesti crijeva. Sadržaj navedenih neesencijalnih elemenata određen je metodom grafitne atomske
apsorpcijske spektrometrije. Srednja vrijednost koncentracije srebra i kositra niža je u mekim
kapsulama, dok se sadržaj barija u tvrdim i mekim kapsulama nije statistički značajno razlikovao.
Količine elementarnih onečišćenja srebra, kositra i barija odgovaraju zahtjevima aktualnih smjernica
ICH Q3 o najvećem dopuštenom dnevnom unosu elementarnog onečišćenja.Inflammatory bowel diseases are affecting an increasing number of patients worldwide. Today, many
patients turn to self-medication and use various dietary supplements. Numerous studies are being
conducted to demonstrate their effectiveness. However, it is crucial to protect patients and ensure
the quality of dietary supplements as prescribed by the guidelines. One of the requirements of these
guidelines is the limitation of elemental impurities, which can have toxic effects on human health.
Therefore, this thesis aimed to determine the content of silver, tin and barium in dietary supplements
for the treatment of inflammatory bowel diseases. The content of the mentioned non-essential
elements was determined using the graphite furnace atomic absorption spectrometry method. The
average concentration of silver and tin is lower in soft capsules, while the barium content in hard
and soft capsules did not differ statistically significantly. The levels of silver, tin and barium
elemental impurities comply with the requirements of the current ICH Q3 guidelines regarding the
maximum permissible daily intake of elemental impurities
Phytochemical characterization of the Melissae folium extracts
No full textMatičnjakov list (Melissae folium) je ljekovita biljna droga koja se temeljem dugogodišnjeiskustvene primjene preporuča kod blagih simptoma stresa, kao pomoć pri spavanju i protiv blažihprobavnih tegoba. Povoljan utjecaj na ljudsko zdravlje pripisuje se sadržaju polifenolnih iterpenskih sastavnica te je stoga važno odrediti njihov sadržaj i sastav u ekstraktima spotencijalnom primjenom u prevenciji i fitoterapiji.
U okviru ovog diplomskog rada provedena je fitokemijska karakterizacija različitih ekstrakatamatičnjakovog lista. U tu su svrhu u vodenim i vodenoetanolnim ekstraktimaspektrofotometrijskim metodama određeni sadržaji ukupnih flavonoida i fenolnih kiselina te jeHPLC-DAD metodom određen sadržaj ružmarinske kiseline kao najzastupljenije polifenolnebioaktivne sastavnice. GC-MS metodom provedena je analiza sastavnica eteričnog uljamatičnjakovog lista. Dobiveni rezultati doprinose znanstvenim spoznajama o bioaktivnimsastavnicama i biomedicinskom potencijalu ekstrakata matičnjaka hrvatskog porijekla.Lemon balm leaf (Melissae folium) is a herbal drug that is, due to experience with its longtraditional use, recommended as medicine for mild mental stress symptoms relief, to aid sleep andmild gastrointestinal complaints. Its beneficial effect on human health is related to the content ofpolyphenolic and terpene compounds and because of that it is important to determine their contentand composition in extracts with the potential of being used in prevention and phytotherapy.
In this thesis a phytochemical characterization of different lemon balm leaf extracts was done.Spectrophotometric methods were used to determine the content of total flavonoids and phenolicacids in aqueous and hydroalcoholic extracts and HPLC-DAD method was used to determine thecontent of rosmarinic acid as a main polyphenolic bioactive compund. GC-MS method was used toanalyse the compounds of the lemon balm leaf essential oil, The obtained results contribute to thescientific knowledges on the bioactive compounds and biomedicinal potential of the lemon balmextracts from Croatia
Application of TXRF-Mo and EDXRF techniques for elemental analysis of whey proteins
No full textProteini sirutke su praškasti oblik proteina dobivenih iz sirutke. Pokazuju brojne korisne učinke na ljudsko zdravlje: antikancerogeni, antioksidativni i antidijabetički učinak, kardioprotektivno i hipotenzivno djelovanje, pozitivno utječu na probavno zdravlje. Na tržištu su dostupni u obliku hidrolizata proteina sirutke, koncentrata proteina sirutke i izolata proteina sirutke. Danas su u širokoj upotrebi pogotovo u fitnes industriji te je zbog toga nužna kontrola elementarnog sastava kako bi se spriječio potencijalno negativan utjecaj na ljudsko zdravlje. Glavni cilj ovog istraživanja je odrediti sadržaj metala u različitim uzorcima proteina sirutke pomoću dvije analitičke tehnike (TXRF-Mo i EDXRF) kao i usporediti učinkovitost primijenjenih tehnika za određivanje sadržaja metala te njihova primjenjivost u kontroli kvalitete proizvoda. Uzorci su pripremljeni mikrovalnom digestijom za potrebe mjerenja TXRF-Mo dok su kod EDXRF tehnike uzorci izravno mjereni bez prethodne pripreme te je određen sadržaj sljedećih elemenata: željezo, bakar, cink, brom, rubidij i stroncij. Dobiveni rezultati pokazali su kako su obje tehnike prikladne za multielementarno određivanje metala te imaju potencijal za određivanje sadržaja broma u uzorcima. Uslijed primijenjenog načina pripreme uzoraka u ovom ispitivanju, EDXRF tehnika je ekološki prihvatljivija i više u skladu s načelima „zelene kemije“ dok je TXRF-Mo osjetljivija tehnika.Whey proteins are a powdered form of protein obtained from whey. They have numerous positive effects on human health: anti-cancer, antioxidant and anti-diabetic effects, cardioprotective and blood pressure lowering effects, positive effects on digestive health. They are available on the market in the form of whey protein hydrolysates, whey protein concentrates and whey protein isolates. Nowadays, they are widely used, especially in the fitness industry, which makes it necessary to control their elemental composition to avoid their potentially negative effects on human health. The main objective of this study is to determine the metal content in different whey protein samples using two analytical techniques (TXRF-Mo and EDXRF) and to compare the effectiveness of the techniques used to determine the metal content and their usefulness in controlling product quality. The samples for the TXRF-Mo measurements were prepared by microwave digestion, while the samples for the EDXRF technique were measured directly without prior preparation. The following elements were determined: iron, copper, zinc, bromine, rubidium and strontium. The results obtained showed that both techniques are suitable for the multi-element determination of metals and have the potential to determine the bromine content in samples. Due to the method of sample preparation used in this test, the EDXRF method proved to be more environmentally friendly and more in line with "green chemistry" principles," while TXRF-Mo is a more sensitive method