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Razvoj robustne metode na osnovi tekočinske kromatografije visoke ločljivosti z masno spektrometrijo za hitro identifikacijo in kvantifikacijo kortikosteroidov v kremah in mazilih
ID Mlakar, Maja (Author), ID Ilaš, Janez (Mentor) More about this mentor... This link opens in a new window

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Abstract
Kortikosteroidi so hormoni, prisotni v telesu, ki zmanjšujejo škodljiv vnetni odgovor organizma. S sintetičnimi kortikosteroidnimi molekulami posnemamo njihovo delovanje in jih uporabljamo za sistemsko ali lokalno zdravljenje kroničnih ali akutnih bolezni, povezanih z vnetjem in bolečino. Za lokalno delovanje na koži vgrajujemo različno aktivne kortikosteroidne molekule v kreme, mazila, losjone ali gele. Topikalne kortikosteroide z nižjo jakostjo uporabljamo za zdravljenje opeklin in alergijskih kožni bolezni, visoko jakostne pa za zdravljenje npr. zadebeljenjih keratotičnih dermatoz. Pri dolgotrajni in prepogosti uporabi takšnih zdravilnih učinkovin in oblik na površino kože pride do reverzibilne ali celo ireverzibilne lokalne atrofije prizadetega dela ter neželenih sistemskih učinkov na hipotalamično-hipofizno-nadledvično os. V magistrski nalogi sem se ukvarjala s perečim problemom ilegalnih proizvodov na trgu, ki naj bi bili izdelani na rastlinski bazi, a vendar vsebujejo sintetične kortikosteroide. Tukaj so predvsem izpostavljeni otroci z večjo površino kože in starostniki, s tanjšo kožno bariero, ki zlahka absorbira vase različne snovi. Nezaveden in tudi nenadzorovan nanos kortikosteroidov na kožo lahko povzroči poslabšanje kožne bolezni zaradi nastanka nepovratnih kožnih atrofij in zavrtja obnove kožnih celic ter oslabljen imunski odziv in posledično razvoj številnih glivičnih okužb. V analizo smo dobili vzorec neznane kreme, ki naj bi bila povsem naravnega, rastlinska izvora, brez prisotnosti kortikosteroidov. Razvili smo enostavno, hitro in robustno HPLC/MS metodo za analizo kreme, s katero smo analite tudi kvantificirali. Pri HPLC-ju smo uporabili reverznofazno kolono in optimizirali metodo za ločbo kortikosteroidnih molekul, ki smo jih imeli na voljo. Z enojnim kvadrupolnim masnim spektrometrom smo ionizirane analite detektirali in posneli masne spektre za posamezne vzorce ter poleg identifikacije z metodo umeritvene krivulje poskušali vsebnost analitov tudi ovrednotiti. Med samim praktičnim delom smo se soočali s problematiko ustrezne priprave vzorca krem predvsem zaradi netopnosti podlage, sam analit pa je bil topen v zmesi acetonitrila in vode. Izvedbo HPLC smo morali časovno podaljšati, da smo dobili ustrezno ločbo vseh predlaganih kortikosteridnih molekul, za kvantifikacijo pa smo izbrali v večini primerov detekcijo adukta molekul z natrijevim ionom. V kremi smo našli kortikosteroid betametazon dipropionat v približno desetkrat nižjih koncentracijah kot ga vsebujejo lekarniške kreme, kar je že zadosti velika količina za neželene učinke molekule s tako močnim delovanjem.

Language:Slovenian
Keywords:kortikosteroidi, kreme in mazila, tekočinska kromatografija visoke ločljivosti, masna speektrometrija
Work type:Master's thesis/paper
Organization:FFA - Faculty of Pharmacy
Year:2019
PID:20.500.12556/RUL-109279 This link opens in a new window
Publication date in RUL:29.08.2019
Views:1353
Downloads:277
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Secondary language

Language:English
Title:Development of a robust method for fast identification and quantification of corticosteroids in creams and ointments based on high-performance liquid chromatography with mass spectrometry
Abstract:
Corticosteroids are hormones present in the body reducing the harmful inflammatory response of the organism. Synthetic corticosteroid molecules are used for the treatment of systemic or local chronic or acute diseases associated with inflammation and pain. For the local application on the skin, we incorporate different corticosteroids of various potency into creams, ointments, lotions or gels. Less potent topical corticosteroids can be used for treating burns and allergic skin diseases, and highly potent ones on the other hand for the treatment of e.g. thickened keratotic dermatoses. If we are applying them on the skin for too long or too frequent, it can lead to reversible or even irreversible local atrophy of the skin and unwanted systemic effects on the hypothalamic-pituitary-adrenal axis. In my master's thesis, I addressed a problem of illegal products on the market, which are supposed to be based on herbs, but actually contain synthetic corticosteroids. More exposed are children who have greater skin area and elderly people, with a thinner, easily absorbing skin barrier. Daily uncontrolled, without supervision application of corticosteroids on the skin can cause a worsening of the skin disease due to the formation of irreversible skin atrophy and the inhibition of skin cell renewal, in addition a weakened immune response can lead to the development of many fungal infections. We have received a sample of unknown cream, which was supposed to be of natural, plant origin, without the presence of corticosteroids. For the analysis of creams we developed a simple, fast and robust method with the HPLC / MS system, with this method we also quantified the analytes. We used a reverse phase column in and optimized the HPLC method for separating the chosen corticosteroid molecules. For individual samples we were detecting the ionized analytes and recorded the mass spectra with a single quadrupole mass spectrometer, and in addition we also evaluated analytes content with the calibration curve method. During the practical work we were faced with the problem of the sample preparation particularly due to the insolubility of the cream base, but the analyte itself was soluble in the mixture of acetonitrile and water. We had to extend HPLC method in order to obtain an appropriate separation of all the proposed corticosteroid molecules, for the quantification we have chosen adduct of sodium ion molecules in most cases. We found a betamethasone dipropionate in tested cream at approximately ten times lower concentrations than are the contents in regular pharmacy creams, which is already sufficient for the adverse effects of such a potent molecule.?

Keywords:corticosteroids, creams and ointments, high resolution liquid chromatography, mass spectrometry

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