| Abstract | Upalne bolesti crijeva spadaju u kronične idiopatske upalne bolesti gastrointestinalnog sustava karakterizirane upalama segmenata probavnog trakta s raznolikim kliničkim manifestacijama poput krvavih stolica, proljeva, tenezma, urgencije te grčevitih bolova. Razlikujemo dva entiteta: Crohnovu bolest i ulcerozni kolitis. U pozadini Crohnove bolesti je stanično posredovani Th1 tip imunog odgovora, a kod ulceroznog kolitisa Th2 tip koji generira humoralni imuni odgovor. Od lijekova koji se koriste za liječenje upalnih bolesti crijeva prevladavaju aminosalicilati, kortikosteroidi, imunosupresivi i biološka terapija, no u posljednje vrijeme sve se više pacijenata uključuje uz terapiju i biljne dodake prehrani. Neki od korištenih dodataka prehrani su ekstrakt andrografisa i smola tamjanovca, no svakako najvažnija je kurkuma u čijim podancima se nalazi spoj kurkumin uz koji se povezuje snažno protuupalno, antioksidativno i antimutageno djelovanje.
S obzirom na to da od 1990-ih postoji jak uzlazni trend u korištenju dodataka prehrani, jasno je da se povećao i interes pacijenata i zdravstvenih radnika za njihovu sigurnost i učinkovitost. U usporedbi s lijekovima, dodaci prehrani nisu podvrgnuti istim znanstvenim ispitivanjima i nisu tako striktno regulirani i kontrolirani, stoga se mogu naći proizvodi kontaminirani s teškim metalima, pesticidima i dodanim farmakološkim tvarima. Kod biljnih dodataka prehrani, najveći problem je što često sadržavaju značajnu količinu zaostalih organskih otapala, posebice etanola koji je metabolički aktivan i kao takav predstavlja rizik za pojedine skupine pacijenata. Dakle, potreban je razvoj jedinstvene analitičke metode koja bi mogla brzo i pouzdano odrediti ostatna otapala prisutna u biljnim dodacima prehrani.
Plinska kromatografija je metoda odjeljivanja u kojoj uzorak tijekom unošenja u kolonu trenutno ispari, nakon čega plinovita pokretna faza eluira sastavnice uzorka prolazeći kolonom sa nepokretnom fazom, a odijeljene se sastavnice najčešće dokazuju plamenoionizacijskim detektorom. Iz uzorka je potrebno prvo odvojiti hlapljiva organska otapala u plinovitu fazu, stoga se plinska kromatografija povezuje s metodom headspace uzorkovanja. S obzirom na hlapljivost organskih otapala i značajnu separacijsku sposobnost kapilarnih kolona, plinska kromatografija vodeća je tehnika za njihovo određivanje.
U ovom radu validirana je sHSS-GC-FID metode za određivanje metanola, etanola, acetona, izopropanola, diklormetana, etil acetata i kloroforma u krutim biljnim dodacima prehrani koji se koriste uz terapiju upalnih bolesti crijeva. Ispitivanje parametara selektivnosti, linearnosti, granica dokazivanja i određivanja, preciznosti, točnosti i izdržljivosti dalo je zadovoljavajuće rezultate, te se metoda pokazala prikladnom za određivanje navedenih ostatnih otapala. Validirana metoda primijenjena je za analizu dva uzorka neobrađene usitnjene kurkume u kojima nije pronađeno nijedno ostatno otapalo, što čini ove biljne dodatke prehrani sigurnima za uporabu u liječenju upalnih bolesti crijeva. |
| Abstract (english) | Inflammatory bowel diseases, which include Crohn's disease and ulcerative colitis, are idiopathic chronical inflammatory diseases of the gastrointestinal tract, characterized by recurrent inflammations of segments of gastrointestinal tract with various clinical manifestations such as blood in the stool, diarrhea, urging and abdominal pain. The pathogenesis of Crohn's disease involves the cell-mediated Th1 immune response, while pathogenesis of ulcerative colitis involves Th2 response which generates humoral immunity. Current drug treatment consists of aminosalicylates, corticosteroids, immunosuppressants and biological therapy, but nowadays more and more patients are turning to herbal therapy because of drug side effects. Some of herbal dietary supplements that are used are extract from green chireta and resin from olibanum-tree, but the most important one is turmeric with its main component curcumin. Numerous pharmacological activities have been reported for curcumin, including anti-inflammatory, antioxidant and anticancer properties.
There is a sharp upward trend in the use of dietary supplements since 1990s, which resulted in widespread interest of health care practitioners and patients in their safety and quality. Comparing to medications, dietary supplements are not subjected to the same scientific scrutiny and are not as strictly regulated and controlled, so they are products sold contaminated with heavy metals, pesticides or prescription drugs. In addition, herbal dietary supplements may contain significant levels of organic volatile compounds, especially ethanol which is metabolically active, so it represents high risk for certain groups of patients. Therefore, development of rapid and reliable method for determination of organic volatile compounds in herbal dietary supplements is much needed.
Gas chromatography is an analytical method in which the sample is loaded onto the head of the column via a heated injection port, where it evaporates, and then a gaseous mobile phase elutes its components flowing through a heated tube coated with a liquid stationary phase. Separated components are usually detected with the flame ionization detector. Gas chromatography may be connected to headspace sampling technique to analyze the samples with complex matrices that also contain non-volatile substances. Due to the volatility of organic solvents and the substantial separating capability of capillary columns, gas chromatography is dominated technique for organic solvents determination.
This work focuses upon validation of sHSS-GC-FID method for simultaneous determination of methanol, ethanol, acetone, isopropanol, dichloromethane, ethyl acetate and chloroform in solid herbal dietary supplements for inflammatory bowel diseases. The method was validated for selectivity, linearity, limits of determination and quantitation, precision, accuracy and robustness. All the results were satisfactory; therefore, the method has proven to be reliable for determination
of these organic volatile compounds. Validated method was used in analysing two different turmeric food products, where none of the organic volatile compounds were found, which has proven their safety in therapeutical use in the inflammatory bowel diseases. |
