Određivanje antocijanina i hidroksicinamske kiseline te njihova količina u različitim vrstama trešnje Prunus avium L. iz Nove Gorice (Slovenija)

The anthocyanins and hydroxycinnamic acids in 5 cultivars of dark coloured sweet cherries were characterised and quantified by means of HPLC and UV-VIS spectrophotometry. Phenolic components were extracted with pure methanol, without addition of acid and water. The samples were diluted in the mixture of methanol and formic acid just before the injection on the column and separated on Hypersil PEP 300 C18 chromatographic column using gradient solvent system consisting of formic acid, water and methanol. DAD detector was employed and two wavelengths were chosen for determination of different components; 320 nm for hydroxycinnamates and 520 nm for anthocyanins. We detected the presence of cyanidin-3-glucoside and cyanidin-3-rutinoside as major anthocyanins, while pelargonidin-3 rutinoside was identified among minor pigments. The major hydroxycinnamic acids were characterised as neochlorogenic acid and 3\u27-p-coumaroylquinic acid. Total anthocyanin content (expressed as cyanidin-3-glucoside) ranged from 29 to 62 mg/100 g of pitted cherry fresh weight (FW), with the highest content observed in Petrovka, a local cultivar. Concentrations of neochlorogenic acid and 3’-p coumaroylquinic acid ranged from 19.5 to 53.0 mg/100 g FW and from 7.5 to 50.6 mg/100 g FW, respectively. The relative amounts of these two phenolic acids varied widely between the cherry cultivars examined in this study.Antocijanini i hidroksicinamske kiseline određeni su i utvrđena je njihova količina HPLC- i UV-VIS-spektrofotometrijom u pet vrsta tamnocrvenih trešanja. Fenolni spojevi ekstrahirani su čistim etanolom bez dodatka kiseline i vode. Uzorci su bili razrijeđeni smjesom metanola i mravlje kiseline neposredno prije injekcije. Na kromatografskoj koloni Hypersil PEP 300 C18 razdvojeni su uzorci korištenjem gradijenta sustava otapala od mravlje kiseline, vode i metanola. Da bi se odredili pojedini sastojci, primijenjen je DAD-detektor i dvije valne duljine, i to: 320 nm za hidroksicinamate i 520 nm za antocijanine. Kao glavni sastojak antocijanina utvr|ena je prisutnost cijanidin-3-glukozida i cijanidin-3-rutinozida, dok je utvrđena manja količina pelargonidina-3-rutinozida. Glavne hidroksicinamske kiseline bile su neoklorogenska i 3´-p-kumaroil-kinska kiselina. Ukupni udjel antocijanina (izražen kao cijanidin-3-glukozid) iznosio je od 29 do 62 mg / 100 g mase svježih plodova bez koštica pri čemu je najveću vrijednost imala vrsta Petrovka. Koncentracija neoklorogenske kiseline iznosila je od 19,5 do 53,0 mg / 100 g svježih trešanja bez koštica, a 3´-p-kumaroil- kinske kiseline 7,5–50,6 mg / 100 g. Relativne količine tih dviju fenolnih kiselina znatno su se razlikovale među pojedinim ispitivanim vrstama trešanja

Determination of polyphenols in white grape berries cv.

Rebula white grapes polyphenols are determined for the first time. We analyzed grapes sampled from 13 different vineyards in Goriška Brda. Total polyphenols were determined according to the Folin-Ciocalteu method. For the individual polyphenols determination we used high performance liquid chromatography (HPLC) -diode array detection (DAD) technique. The separation of polyphenols from grapes was performed using narrow bore C18 Luna column (Phenomenex, 2 x 150 mm, 3µm) and monitored at 320 nm. The majority of white grapes polyphenols was represented by four hydroxycinnamic acids (HCAs). According to their UV-Vis spectra and chromatographic retention properties they corresponded to trans-caftaric, trans-coutaric, cis-coutaric and trans-fertaric acid. To confirm the identity of separated polyphenols, they were also directed to the ion trap mass spectrophotometer (Finnigan LCQ Deca) fitted with electrospray ionization (ESI) probe. Spectra were recorded in negative mode between m/z 100 and m/z 500. The mass spectrometer was programmed to do two consecutive scans: a full mass (MS) and an MS 2 scan of the most abundant ion in the full mass spectra. The results confirmed the assumptive identification of three HCAs; trans-caftaric, trans-coutaric and trans-fertaric acids in white grape berries cv. Rebula based on HPLC-DAD analysis

CLINICAL RECOMMENDATIONS FOR DIAGNOSING, TREATMENT AND MONITORING OF PATIENTS WITH UTERINE CERVICAL CANCER – CROATIAN ONCOLOGY SOCIETY AND CROATIAN SOCIETY FOR GYNECOLOGY AND OBSTETRICS AS CROATIAN MEDICAL ASSOCIATION UNITS AND CROATIAN SOCIETY OF GYNECOLOGICAL ONCOLOGY

Rak vrata maternice, u odnosu na malignome drugih ginekoloških sijela, jest bolest mlađih žena koja se može redovitim kontrolama i zdravstvenim odgojem prevenirati, a u slučaju pojave bolesti učinkovito liječiti. Metode liječenja uključuju kirurgiju, radioterapiju i kemoterapiju, ovisno o stadiju bolesti i općem stanju bolesnica. Odluku o liječenju donosi multidisciplinarni tim. S obzirom na važnost ove bolesti, potrebno je definirati i provoditi standardizirani pristup u dijagnostici, liječenju i praćenju ovih bolesnica. U tekstu koji slijedi iznesene su kliničke smjernice s ciljem implementacije standardiziranih postupaka u radu s bolesnicama s rakom vrata maternice u Republici Hrvatskoj.Cervical cancer, in comparison with other gynecological malignancies, mainly affects younger women. It can be prevented trough educational programs, screening and early detection. It also can be efficiently treated when it appears. Treatment modalities include surgery, chemotherapy and radiotherapy, according to the stage of the disease and patient condition. Treatment decisions should be made after multidisciplinary team discussion. Due to the significance of this disease it is important to define and implement standardized approach for diagnostic, treatment and monitoring algorithm as well. The following text presents the clinical guidelines in order to standardize the procedures and criteria for the diagnosis, management, treatment and monitoring of patients with uterine cervical cancer in the Republic of Croatia

Identification of sweet cherry anthocyanins and hydroxycinnamic acids using HPLC coupled with DAD and MS detector

In absence of standards, HPLC coupled with DAD offers identification of polyphenols by scanning UV-Vis spectra of individual components, which spectral characteristics are unique, but not selective. At the same time HPLC-DAD determination methods of polyphenols differ in mobile phase solutions resulting in DAD scanned spectra deviation between different studies, aggravating the precise identification based on agreement to UV-Vis data from literature. Mass spectrometry (MS) detection with molar weight determination of the individual components in the sample enables more precise identification of compounds eluted from the column. Sweet cherry Petrovka polyphenols were separated on C18 Hypersil PEP 300 column (250 x 4.6 mm, 5 [mu]m) using gradient solvent mixture consisting of methanol, water and formic acid. MS and UV-Vis spectra of eluted anthocyanins and hydroxycinnamic acid were recorded. HPLC-MS analyses were performed using a LCQ [sup]TM ion trap, Finnigan, MAT mass spectrometer by atmospheric pressure chemical ionisation (ACPI). Molecular and fragmented ion masses of sweet cherry hydroxycinnamic acids and anthocyanins were determined and with UV-Vis spectra, in the range of 190-600 nm, used for identification of compounds. Electro spray mass spectrum of two hydroxycinnamic acids produced ions with m/z ratios of 353.0 and 337.0, which corresponded to molecular weights of neochlorogenic acid and 3\u27-p-coumarylquinic acid. The molecular weights of 5 anthocyanins corresponded to cyanidin-3-glucoside (449.0), cyanidin-3-rutinoside (595.1), peonidin-3-glucoside (463.1), pelargonidin-3-rutinoside (579.1) and peonidin-3-rlltinoside (609.2)

Determination and Quantitation of Anthocyanins and Hydroxycinnamic Acids in Different Cultivars of Sweet Cherries (Prunus avium L.) from Nova Gorica Region (Slovenia)

The anthocyanins and hydroxycinnamic acids in 5 cultivars of dark coloured sweet cherries were characterised and quantified by means of HPLC and UV-VIS spectrophotometry. Phenolic components were extracted with pure methanol, without addition of acid and water. The samples were diluted in the mixture of methanol and formic acid just before the injection on the column and separated on Hypersil PEP 300 C18 chromatographic column using gradient solvent system consisting of formic acid, water and methanol. DAD detector was employed and two wavelengths were chosen for determination of different components; 320 nm for hydroxycinnamates and 520 nm for anthocyanins. We detected the presence of cyanidin-3-glucoside and cyanidin-3-rutinoside as major anthocyanins, while pelargonidin-3 rutinoside was identified among minor pigments. The major hydroxycinnamic acids were characterised as neochlorogenic acid and 3'-p-coumaroylquinic acid. Total anthocyanin content (expressed as cyanidin-3-glucoside) ranged from 29 to 62 mg/100 g of pitted cherry fresh weight (FW), with the highest content observed in Petrovka, a local cultivar. Concentrations of neochlorogenic acid and 3’-p coumaroylquinic acid ranged from 19.5 to 53.0 mg/100 g FW and from 7.5 to 50.6 mg/100 g FW, respectively. The relative amounts of these two phenolic acids varied widely between the cherry cultivars examined in this study