Pharmacy Practice and Education in Finland.

The Pharmacy Education in Europe (PHARMINE) project studies pharmacy practice and education in the European Union (EU) member states. The work was carried out using an electronic survey sent to chosen pharmacy representatives. The surveys of the individual member states are now being published as reference documents for students and staff interested in research on pharmacy education in the EU, and in mobility. This paper presents the results of the PHARMINE survey on pharmacy practice and education in Finland. Pharmacies have a monopoly on the dispensation of medicines. They can also provide diagnostic services. Proviisori act as pharmacy owners and managers. They follow a five-year (M.Sc. Pharm.) degree course with a six-month traineeship. Farmaseutti, who follow a three-year (B.Sc. Pharm.) degree course (also with a six-month traineeship), can dispense medicines and counsel patients in Finland. The B.Sc. and the first three years of the M.Sc. involve the same course. The current pharmacy curriculum (revised in 2014) is based on five strands: (1) pharmacy as a multidisciplinary science with numerous opportunities in the working life, (2) basics of pharmaceutical sciences, (3) patient and medication, (4) optional studies and selected study paths, and (5) drug development and use. The learning outcomes of the pharmacy graduates include (1) basics of natural sciences: chemistry, physics, technology, biosciences required for all the students (B.Sc. and M.Sc.), (2) medicine and medication: compounding of medicines, holism of medication, pharmacology and biopharmaceutics (side-effects and interactions), patient counseling, efficacy and safety of medicines and medication, (3) comprehensive and supportive interactions of the various disciplines of pharmacy education and research: the role and significance of pharmacy as a discipline in society, the necessary skills and knowledge in scientific thinking and pharmaceutical research, and (4) basics of economics and management, multidisciplinarity, hospital pharmacy, scientific writing skills, management skills. In addition, teaching and learning of "general skills", such as the pharmacist's professional identity and the role in society as a part of the healthcare system, critical and creative thinking, problem-solving skills, personal learning skills and life-long learning, attitude and sense of responsibility, and communication skills are developed in direct association with subject-specific courses. Professional specialization studies in industrial pharmacy, and community and hospital pharmacy are given at the post-graduate level at the University of Helsinki.

Competence-Based Pharmacy Education in the University of Helsinki.

In order to meet the expectations to act as an expert in the health care profession, it is of utmost importance that pharmacy education creates knowledge and skills needed in today's working life. Thus, the planning of the curriculum should be based on relevant and up-to-date learning outcomes. In the University of Helsinki, a university wide curriculum reform called 'the Big Wheel' was launched in 2015. After the reform, the basic degrees of the university are two-cycle (Bachelor-Master) and competence-based, where the learning outcomes form a solid basis for the curriculum goals and implementation. In the Faculty of Pharmacy, this curriculum reform was conducted in two phases during 2012-2016. The construction of the curriculum was based on the most relevant learning outcomes concerning working life via high quality first (Bachelor of Science in Pharmacy) and second (Master of Science in Pharmacy) cycle degree programs. The reform was kicked off by interviewing all the relevant stakeholders: students, teachers, and pharmacists/experts in all the working life sectors of pharmacy. Based on these interviews, the intended learning outcomes of the Pharmacy degree programs were defined including both subject/contents-related and generic skills. The curriculum design was based on the principles of constructive alignment and new structures and methods were applied in order to foster the implementation of the learning outcomes. During the process, it became evident that a competence-based curriculum can be created only in close co-operation with the stakeholders, including teachers and students. Well-structured and facilitated co-operation amongst the teachers enabled the development of many new and innovative teaching practices. The European Union funded PHAR-QA project provided, at the same time, a highly relevant framework to compare the curriculum development in Helsinki against Europe-wide definitions of competences and learning outcomes in pharmacy education.

Feasibility of SU-8-based capillary electrophoresis-electrospray ionization mass spectrometry microfluidic chips for the analysis of human cell lysates.

Monolithically integrated, polymer (SU-8) microchips comprising an electrophoretic separation unit, a sheath flow interface and an ESI emitter were developed to improve the speed and throughput of proteomics analyses. Validation of the microchip method was performed based on peptide mass fingerprinting and single peptide sequencing of selected protein standards. Rapid, yet reliable identification of four biologically important proteins (cytochrome C, ß-lactoglobulin, ovalbumin and BSA) confirmed the applicability of the SU-8 microchips to ambitious proteomic applications and allowed their use in the analysis of human muscle cell lysates. The characteristic tryptic peptides were easily separated with plate numbers approaching 10(6), and with peak widths at half height as low as 0.6 s. The on-chip sheath flow interface was also exploited to the introduction of an internal mass calibrant along with the sheath liquid which enabled accurate mass measurements by high-resolution Q-TOF MS. Additionally, peptide structural characterization and protein identification based on MS/MS fragmentation data of a single tryptic peptide was obtained using an ion trap instrument. Protein sequence coverages exceeding 50% were routinely obtained without any pretreatment of the proteolytic samples and a typical total analysis time from sampling to detection was well below ten minutes. In conclusion, monolithically integrated, dead-volume-free, SU-8 microchips proved to be a promising platform for fast and reliable analysis of complex proteomic samples. Good analytical performance of the microchips was shown by performing both peptide mass fingerprinting of complex cell lysates and protein identification based on single peptide sequencing.

Glucuronidation of racemic O-desmethyltramadol, the active metabolite of tramadol.

O-Desmethyltramadol, the active metabolite of analgesic tramadol, is metabolised through glucuronidation. The present study was conducted to identify the human UDP-glucuronosyltransferases (UGTs) that catalyse the glucuronidation of O-desmethyltramadol, a racemic mixture of 1R,2R- and 1S,2S-enantiomers. We developed a fast and selective liquid chromatography-mass spectrometry method to separate, analyse and quantify the diastereomeric phenolic O-glucuronides of O-desmethyltramadol. To quantify O-desmethyltramadol glucuronidation, we biosynthesised both phenolic O-glucuronides of O-desmethyltramadol and verified their structure by mass spectrometry and nuclear magnetic resonance spectroscopy. Subsequently, the 16 human UGTs of subfamilies 1A and 2B were screened for O-desmethyltramadol glucuronidation activity. UGTs 1A7-1A10 exhibited a strict stereoselectivity, exclusively glucuroniding the 1R,2R-enantiomer. Similar though not strict enantioselectivity was exhibited by UGT2B15. UGT2B7, on the other hand, glucuronidated both O-desmethyltramadol enantiomers, with slight preference for 1S,2S-O-desmethyltramadol. Enzyme kinetic parameters were determined for the most active UGTs, 1A8 and 2B7. The apparent K(m) or S(50) values were high: 1.2mM±0.23 for 1R,2R-O-desmethyltramadol with UGT1A8 and 1.84±1.2 and 4.6±2.0mM for 1S,2S- and 1R,2R-O-desmethyltramadol enantiomers with UGT2B7, respectively. Glucuronidation analyses of O-desmethyltramadol with human liver microsomes exhibited stereoselectivity, favouring the 1S,2S-O-desmethyltramadol over 1R,2R-O-desmethyltramadol and yielding 62.4 and 24.6pmol/mg/min, respectively. In intestinal microsomes, on the other hand, the two enantiomers were glucuronidated at similar rates, about 6pmol/mg/min. The results shed new light on both tramadol metabolism and the substrate selectivity of the human UGTs.

Simple coupling of gas chromatography to electrospray ionization mass spectrometry.

A simple method for direct coupling of gas chromatography (GC) with electrospray ionization mass spectrometry (ESI/MS) has been developed. The outlet of the GC capillary column was placed between the ESI needle and the atmospheric pressure ionization (API) source of a mass spectrometer. The ionization occurs via dissolution of neutral compounds into the charged ESI droplet followed by ion evaporation or via a gas-phase proton transfer reaction between a protonated solvent molecule and an analyte. The mass spectra of organic volatile compounds showed abundant protonated molecules with little fragmentation, being very similar to those produced by normal liquid ESI. The quantitative performance of the system was evaluated by determining the limit of detection (LOD), linearity ( r (2)), and repeatability (RSD). The GC-ESI/MS method was shown to be stable, providing high sensitivity and good quantitative performance.

Determination of dopamine and methoxycatecholamines in patient urine by liquid chromatography with electrochemical detection and by capillary electrophoresis coupled with spectrophotometry and mass spectrometry.

The applicability of capillary electrophoresis (CE) with UV and mass spectrometric (MS) detection for the determination of dopamine and methoxycatecholamines in urine was evaluated in comparison with the liquid chromatography-electrochemical detection (LC-EC) method widely used in catecholamine analysis. The catecholamines in urine were deconjugated with acid or enzyme hydrolysis, purified by cation exchange (CEX) or solid-phase extraction (SPE) with a copolymer of N-divinylpyrrolidone and divinylbenzene and analyzed by LC-EC, CE-UV, and CE-MS. Acid hydrolysis was more effective in the deconjugation than enzymatic hydrolysis with Helix pomatia. However, the recoveries of HMBA, DA and NMN from spiked samples were less than 30% after acid hydrolysis and SPE purification. The CEX purification was more efficient than SPE in removing matrix compounds from the urine samples. The limits of detection were lower in LC-EC analysis than in CE-UV or CE-MS. Many factors in the analytical procedure caused deviations in the concentrations measured for urinary dopamine and methoxycatecholamines. The recovery of HMBA, which was used as the internal standard, was poor after acid hydrolysis and SPE purification. The purification methods were validated in conjunction with the analytical methods and therefore cross analysis was unsuccessful. The LC-EC method was the most sensitive, but CE-UV and CE-MS were sensitive enough for the determination of dopamine and methoxycatecholamines even in healthy patient urine. The EC and MS detections were superior to the UV detection in specificity since, after acid hydrolysis, some matrix compounds were migrating close to I.S., DA and 3MT.

Analysis of catecholamines by capillary electrophoresis and capillary electrophoresis-nanospray mass spectrometry. Use of aqueous and non-aqueous solutions compared with physical parameters.

Catecholamines were analysed in aqueous and alcoholic non-aqueous solutions by capillary electrophoresis and capillary electrophoresis-mass spectrometry using sheathless nanospray coupling. Decreases in the electrophoretic mobilities of the catecholamines and in the electroosmotic mobilities were observed from water to 1-propanol. Separations were more efficient in all non-aqueous media than in water. The diffusion coefficients of the catecholamines in the different media were determined. The solvent had little effect on the sensitivity of the UV or MS detection. Both methods were successfully applied to the analysis of urine samples.