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The contamination of botanical dietary supplements with pesticide and fungicide ethylene oxide (EO) is well-recognized problem worldwide. Contamination of botanicals with pathogenic bacteria is attributed in part to growing conditions and environment, sanitation and hygiene practices among harvest workers, and lack of good agricultural and manufacturing practices within some developing countries. Processing methods for botanicals include fumigation with EO, irradiation, and vacuum-assisted steam. Moreover, capsule shells as the most used delivery system of botanical dietary supplements can be sterilized with EO by manufacturers before releasing to user enterprises. [1, 2] Due to its mutagenic and carcinogenic properties, herbal dietary supplements need to comply with the maximum residue level established by regulatory agencies. [3, 4] It is important to ensure that the levels of residual EO and its by-product 2-chloroethanol (2-CE) obtained by the reaction of acetaldehyde and chloride ion pose a minimal risk to the patient. The aim of this research was the assessment of EOTotal expressed as EO equivalents as the sum of native EO and 2-CE in 53 samples of dietary supplements used in the treatment of inflammatory bowel disease (IBD). 28 samples were obtained from local pharmacies and health food stores and 26 were purchased from the Internet. The dietary supplements analyzed in this work were in multiple oral dosage forms including tablets (6), hard capsules (25), soft capsules (4), powders (16) and liquid extracts (2). The main botanical ingredients included turmeric, Indian frankincense and green chiretta. The samples were analyzed using GC-MS/MS (model TQ8050, Shimadzu). In 25 samples (43%) EO and 2-CE were quantified in the range from 0.02 to 3.29 mg/kg. 2 samples (1 hard capsule and 1 tablet) contained levels higher than 1 mg/kg. This study provides a reference for regulatory bodies to routinely control the quality of botanical dietary supplements and to prevent EO residues from jeopardizing patients’ health.

As the number of patients with inflammatory bowel disease (IBD) that use dietary supplements (DS) continues to increase in many countries, the importance of ensuring product quality is of scientific matter. Regulators, health professionals, and manufacturers often disagree on how much quality testing is necessary for DS. This is highlighted by the World Health Organization’s Strategy on Traditional Medicines 2014–2023 [1] where quality is seen as a basis of the DS sector. The United States Pharmacopeia (USP) is an independent, nonprofit, science-based organization whose mission is to improve human health through quality standards and tests for DS. [2] The aim of this research was to perform oral dosage testing based on USP protocols of botanical DS used in the treatment of IBD. 53 samples tested were in multiple oral dosage forms: tablets, hard capsules, soft capsules, powders, and liquid extracts. The following test were performed: Tapped Density , Loss on Drying , Tablet Friability , Tablet Breaking Force , Weight Variation , and Disintegration . The tests were performed using tapped density testing instrument (PT-DT300), tablet friability test instrument (PTF 100), and tablet disintegration tester (PTZ-S) by Pharma Test (Germany), tablet hardness tester (TBH 125) by Erweka (Germany) and moisture analyzer (DBS 60-3) obtained by Kern & Sohn GmbH (Germany). The lowest tapped density was found for turmeric (CI 25%) and the highest for green chiretta (CI 45%). The tablet breaking force was in the range of 27.7 to 248.5 N. One sample containing black pepper as a botanical ingredient did not meet the requirements of loss on the drying test (5.2%). All investigated tablets met the requirements of the friability test expressed in terms of allowable weight loss (≤0.61%). Investigated tablets and capsules met the requirements of the weight variation test expressed in terms of the allowable deviation from the average weight of the sample (≤7.7% tablets, ≤9.0% capsule). Only one sample did not meet the requirements of the disintegration test as none of investigated 6 tablets had not disintegrated under specified conditions. The challenges in DS science and its regulation provide new opportunities for scientists and regulators to work together both nationally and internationally, and to cooperate to improve public health. Assessments of the quality of DS are needed to provide the scientific information that regulators need.

Inflammatory bowel disease is a complex disorder characterized by chronic gastrointestinal inflammation. Thus, patients prefer to use herbal dietary supplements containing turmeric, Indian frankincense, green chiretta, and black pepper in an attempt to cope better with their chronic condition. The dietary supplements’ dosage forms and herbal ingredients were assessed in terms of the products’ physicochemical parameters (weight uniformity, friability, disintegration, rupture test, tablet’s breaking force, and powder flowability) in view of the USP-NF requirements. In addition, contaminants such as organic solvents and ethylene oxide were evaluated using gas chromatography. Assessment of gluten via an Enzyme-Linked Immunosorbent Assay was also performed. Most of the products met USP requirements. The high average weight of one multicomponent tablet sample with a high breaking force value can explain the observed negative results of the disintegration test. A total of 26% of samples tested positive for gluten, but the most alarming fact is that the ethylene oxide levels found in two samples were up to 30 times higher than the EU limit. Accordingly, dietary supplement quality control is of fundamental importance.

U ovom preglednom radu opisana je tehnika ekstrakcije čvrstom fazom te je prikazana njena primjena u bioanalitici. Rad obuhvaća pregled bioanalitičkih metoda namijenjenih određivanju terapijske doze lijekova te ispitivanju njihove farmakokinetike. Nadalje, opisane su bioanalitičke metode kojima se određuju biomarkeri različitih bolesti, opijatne droge, sintetički stimulansi i štetne tvari koje se unose prehrambenim i kozmetičkim proizvodima. Navedeni su raznovrsni reaktivni sorbensi od široko komercijalno dostupnih modificiranih silikagelova i polimera do visoko selektivnih inovativnih sorbensa poput nanomaterijala, polimera s molekulskim otiskom, imunosorbensa itd. Osim seruma, plazme i urina, istaknute su i metode kojima su analizirane druge vrste uzoraka poput sline, bronhalnog sekreta, koštane srži, majčinog mlijeka, mekonija i kose. Automatizacija bioanalitičkih metoda je danas imperativ, posebice u kliničkim laboratorijima stoga su obrađene i metode koje koriste formate ekstrakcije čvrstom fazom namijenjene analizi velikog broja uzoraka poput 96-ekstrakcijskih pločica i on-line kolona.

Biomimetička kromatografija danas ima sve važniju ulogu u razvoju lijekova. !mobilizacija raznih bioloških struktura poput humanog serumskog albumina, al-kiselog glikoproteina, fosfolipida te raznih enzima i receptora na nepokretne faze kromatografske kolone dovela je do mogućnosti ispitivanja velikog broja spoj eva u kratkom vremenu. Biomimetičkom kromatografijom mogu se ispitati parametri poput vezanja za proteine plazme, lipofilnosti te volumena raspodjele, kao i afiniteta za određene mete ili receptore. Na taj način moguće je dobiti uvid u farmakokinetička i farmakodinamička svojstva velikog broja spoj eva kandidata za lijek te odabrati najpogodnije za nastavak istraživanja, time ubrzavajući proces razvoja lij ekova.

Sve veći broj ljudi boluje od upalnih bolesti crijeva (engl. Inflammatory Bowel Disease, IBD), kojima je jedan od najčešćih simptoma malapsorpcija hranjivih tvari poput folne kiseline (engl. folic acid, FA, vitamin B9) čiji je deficit povezan s povećanim rizikom karcinoma kolona. U svrhu povećanja adherencije pacijenata predlaže se dvokomponentna formulacija jednog od imunosupresiva korištenog u terapiji IBD: azatioprina (AZA), 6- merkaptopurina (6-MP) ili 6-tiogvanina (6-TG) te FA. Cilj istraživanja je razviti metodu koja bi predstavljala analitičku podršku predloženim formulacijama. Analiza uzoraka provedena je korištenjem tehnike tekućinske kromatografije visoke djelotvornosti spregnute s dvojnom spektrometrijom masa (LC- DAD-MSn). Korištena je Zorbax SB-C8 kolona te acetonitril i ultračista voda zakiseljeni mravljom kiselinom kao pokretna faza, uz gradijentni program. Razvojem metode optimirani su različiti parametri metode poput pokretne i nepokretne faze, ekstrakcijskog otapala (MeOH : 0, 1 M NaOH, 50 : 50, V : V), internog standarda (2- amino-6-metilmerkaptopurin) uz trajanje analize od 10 min. Metoda je validirana prema ICH smjernicama, pri čemu pokazuje izvrsnu linearnost, točnost i preciznost te je primijenjena na tabletama uz analitičke prinose od 90, 17 do 99, 66%. U ovom je istraživanju razvijena, validirana i primijenjena LC-DAD-MSn metoda za istovremeno određivanje AZA, 6-MP, 6- TG i FA u tabletama. S obzirom na kratko trajanje analitičkog postupka, metoda pokazuje potencijal korištenja u rutinskim analizama. Ovaj rad je sufinancirala Hrvatska zaklada za znanost projektom UIP-2017-05-3949.

In recent years, in most developed countries, the use of dietary supplements in general, and especially those intended for children is increasing. Despite the fact that most dietary supplements, especially those intended for children, can have side effects, legislation puts them in a group of foods, not drugs. Therefore there are not subject to such strict requirements of safety, quality and efficiency as medicines are. Because of higher availability, but also because of many other reasons, such as growing increase of self-care in general and growing concern of modern man for their own health, the opinion is prevailing that dietary supplements are safe. It is undisputed that the majority of food supplements for children can have very beneficial effects on their overall health. However, not enough attention is given to the fact that dietary supplements may have a positive effect only if these products are used for the right reason at the right time, in the right dose and in the right way. Do to the fact that children are a special group that are particularly vulnerable, one must keep in mind the issue of production and distribution practice, and the use of food supplements should be approached with caution. In order to avoid a number of potential risks that the use of dietary supplements in general especially those intended for children brings, dietary supplements should be subject to continuous scientific research, pharmaceutical expert supervision and appropriate legal controls. It is extremely important that the parents themselves acquire more knowledge about dietary supplements used by their children. Only if all above mentioned criteria are met, dietary supplements can achieve their optimum performance. The aim of this survey was to determine how much parents are familiar with dietary supplements intended for their children. In addition to evaluating the level of their knowledge on the composition, purpose and method of use of dietary supplements, in the present study we investigated the attitudes and opinions that parents generally have about dietary supplements. Based on the data of the survey conducted on a sample of parents (N = 415) it was found that parents perceive dietary supplements as safe. The research results unequivocally confirm the disturbingly low level of awareness of parents about dietary supplements for children. Such insight is ultimately a claim for an important advisory role of a pharmacist. In fact, the pharmacist should play a key role in advising parents to make an informed decision on the selection and manner of use of the food supplement for their child. Based on their knowledge, experience and expert assessment, pharmacists will decide whether there is a need for a certain dietary supplement, depending on the intended use and indications. If necessary they will recommend a different dietary supplement, explain in what cases to use it, and for how long, as well as how to use it. The pharmacist must raise parents’ awareness that dietary supplements contain active substance and that they should not be taken lightly and that the recommended daily dose should not be exceed. It is also necessary to point out that dietary supplements for children, especially those that contain plant extracts, may contain alcohol. The pharmacist, when dispensing medicines, should make a recommendation what dietary supplement may contribute to faster recovery, prevent or mitigate side effects and complications of pharmacotherapy. It is also important that the pharmacist emphasizes that certain dietary supplements should not be used with a certain drug because of possible interactions. In the end, it is necessary to point out that the pharmacist should play a key role in advising parents to make an informed decision on the selection of the dietary supplement for their child as well as the correct way of using it.

A cough syrup is a medicinal product used in an attempt to treat coughing and related conditions. Traditional herbs are used for preparation of cough syrup. There are two main categories of herbs that are used in preparation of cough remedies: expectorants and antitussives. For productive coughs, treatment with expectorants may be attempted to loosen mucus from the respiratory tract. For dry coughs antitussives are used to suppress the body’s urge to cough. Herbal medicinal products may contain significant levels of ethanol arising from its use as an extraction solvent in liquid extracts and tinctures. Although the use of ethanol is necessary for extraction of pharmacologically active compounds, it was found that some herbal medicinal products contain large amounts (up to 60 %) of ethanol with even less clinical need or benefit. In young children ingestion of even modest amounts of ethanol may lead to permanent neurological damage as result of hypoglycemia. Therefore, special attention should be given to the ethanol content of herbal medicinal products for pediatric use. At the moment the safety evaluation of the ethanol content of herbal medicinal products for pediatric use is not harmonized between different European Union Member States. Moreover, the lack of guidelines relating to safe limits of ethanol as part of herbal medicinal products for pediatric use has also led to different national labeling practices. Therefore, the aim of our work was to develop and validate a new HSS-GC-FID method for quantification of ethanol and its main impurity, methanol, present in children’s cough syrup.

Red fermented rice, also known as red yeast rice, Hongqu, Anka or red Koji, is the fermented product of rice on which red yeast (Monascus purpureus) has been grown. For a thousand of years it has been used in China to make rice wine, as a food preservative for maintaining the color and taste of fish or meat and as a traditional medicine for digestive and vascular function. A complete and detailed description of its manufacture and medicinal properties are found in the ancient Chinese pharmacopoeia, published during the Ming Dynasty (1368–1644 a.d.). Over the past decade, there has been increased scientific interest in red yeast rice, as an alternative therapy for hyper cholesterolemia. Many studies have demonstrated that red yeast rice may be especially useful for treating patients who refuse to take statin drugs because of philosophical reasons or patients who are unable to tolerate statin therapy due to statin-associated myalgias. The major components found in red yeast rice, known as monacolins (polyketides), are reported to exhibit a cholesterol lowering action by inhibiting the 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase. Still, it is believed that the hypolipidemic effect of red yeast rice cannot be accounted for only by the monacolins, but by a synergy of all other components present in red yeast rice: plant sterols, isoflavones and isoflavone glycosides, selenium and zinc. The most important toxic component in red fermented rice is citrinin, a mycotoxin produced by red yeast (Monascus purpureus). It has been known to cause kidney and liver damage in humans. Therefore, Japan has issued an advisory limit of 200 ppb while the European Union has a recommended limit of 100 ppb. The aim of our work was to develop a new LC/MS/MS method for identification of active components, monacolin K (lovastatin) and monacolin K acid and toxic component, citrinin, present in red yeast rice capsules.

Artichoke (Cynara scolymus), belonging to the family of Asteraceae, is an herbaceous perennial crop, widely cultivated in the Mediterranean area. The heads of the artichoke are edible and used worldwide, while the leaves are an herbal medicine recognized far a long time far their beneficial and therapeutic effects, including induction of choleresis, LDL oxidation and significant antioxidant, antifungal, antibacterial as well as strong hepatoprotective activity. Artichoke contains very little fat and high levels of minerals, vitamins, fibre, inuline and polyphenols. The most abundant phenolic compounds reported in artichoke leaf are flavonoides such as apigenin and luteolin (both glycosides and rutinosides), as well as phenolic acids, particularly derivatives of mono- and di-caffeoylquinic acid. Although all polyphenols are included in cholesterol lowering effect of artichoke leaf, the most responsible one is cynarin, derivative of dicaffeoylquinic acid. As dietary supplements have an important position in health care systems worldwide, their current assessment and quality control are a major bottleneck. Over the last years, major steps were taken to improve the quality of the dietary supplements and also to develop analytical methods ensuring their quality. High performance liquid chromatography has become the most popular analytical technique far analyzing dietary supplements. The main advantages of the technique are high resolution, selectivity, sensitivity and possibility to couple the technique to different detectors. Therefore, the aim of the present work was to develop a new HPLC! DADIMS/MS method far identification of cynarin present in artichoke leaf extract tablets.

Blackberries are grown in continental Croatia on small farms using traditional techniques of cultivation and harvesting by hand in summers. The most common blackberry cultivars in Croatia are Thornless Lagan, Thornfree, Black Satin and Tayberry. A significant proportion of fresh fruits is directly processed into blackberry products · jam, juice or wine. Blackberry wine is a product of yeast fermentation of natural sugars present in blackberry juice. In addition to being a rich source of vitamins and dietary fibre, blackberry wine is rich in phenolic compounds such as anthocyanins, flavonols, flavanols, proanthocyanidins, and phenolic acids. During fermentation of blackberries, yeast interact with sugars in the juice to create ethanol, while abundance of other alcohols should be significantly lower. Methanol is a highly toxic alcohol whose ingestion or inhalation can cause damage of central nervous system, blindness and even death. In the view of this, it is extremely important to regularly control methanol content in the blackberry wines. Therefore, the aim of our work was to develop a new headspace gas chromatographic method with flame ionization detection (HSS-GC-FID) far the determination of methanol as the main volatile impurity present in blackberry wines.

Molecular lipophilicity was studied using salicylamide as a model drug. Log P value for the target compound was experimentally determined by "shake-flask" method and calculated using nine different computer programs: based on atom contributions, based on atom/fragment contributions, based on 13 structural parameters, based on fragmental contributions, based on atom-type electrotopological-state indices and neural network modeling, and based on topological structure descriptors. Our analysis demostrates perfect agreement between experimentally observed log P value of salicylamide and calculated value by CSLogP program, based on topological structure descriptors and electrotopological indices.

Flavonoids are one of the largest groups of natural compounds known. They are reputated to have numerous physiological activities. There are many food supplies which contain flavonoids, but one of the most important source is propolis. In propolis, main active substances except flavonoids are phenolic acids. Nowdays, the use and list of preparations of propolis became nearly endless, and demands an accurate analytical method to define the substances in this natural product. It is easily analysed by chromatographic methods, but before testing some new type of propolis it is better to optimize chromatographic conditions. The aim of this study is to optimize chromatographic conditions in TLC of flavonoids and phenolic acids as standard compounds that may be present in Croatian propolis. We used 9 different mobile phases and using information theory and numerical taxonomy methods compared by application of computer search program KT1, we found the most appropriate mobile phase, the optimal combination of two and three mobile phases for separation of standards.

In our work log P value for 3, 3´-(2-metoxybenzilidene)bis[4-hydroxycoumarin] (MBbisHC) was experimentally determinated (˝shake-flask˝ method) and calculated using six computer programs: HyperChem 7.0 (based on atom contributions), XLOGP (based on atom contributions), KowWin (based on atom/fragment contributions), CLOGP (based on fragmental contributions), ALOGPS 2.1 (based on atom-type electrotopological-state indices and neural network modeling), and IA logP (based on atom-type electrotopological-state indices and neural network modeling). The experimental and calculated log P values were correlated. There is nearly perfect agreement between experimentally observed and calculated log P (HyperChem 7.0) value.

In this work, we described the LogKow program that estimafe-s the log octanol/water partition coefficient (log P) of organic chemicals using a atorn/fragmental method developed at SRC. Chemical structures can be brought into LogKow estimation program using SMILES notation, CAS RN or commercial drawing programs that allow a user to draw a chemical structure and then directly write the SMILES notation. SMILES is an acronym for Simplified Molecular Input Line Entry System. It is a chemical notation system used to represent a molecular structure by a linear string of symbols. A two-dimensional drawing of a single chemical structure is possible in many different forms. That is, a single structure can be depicted correctly by many different SMILES notations. SMILES notations are comprised of atoms (designated by atomic symbols), bonds, parentheses (used to show branching), and numers (used to designate ring opening and closing positions). The SMILECAS Database is very helpful and time-efficient in obtaining SMILES notations and log P values; all that you need is CAS Reigistry Number. This database contains the SMILES notations for 103 000 compounds. In order to evaluate this method estimated log P values were compered with these experimental obtained from the SMILECAS Database. Statistical analysis has been performed using the StatSoft Inc.1992. Microsoft Corp. program. Results show that there was no statistical significant difference, p