Category Archives: AMM 2019, Volume 65, Number 2

The Statistical Analysis of Pharmacokinetic Parameters in the Context of Bioequivalence Testing of Two Anthelmintic Formulas Based on Ivermectine and Triclabendazole in Sheep

DOI: 10.2478/amma-2019-0015

Conducting bioequivalence studies is an essential step during the market authorization process of generic pharmaceutical formulations, for both human or veterinary use. The aim of the present study was to evaluate the pharmacokinetics of triclabendazole sulphoxide, the main metabolite of triclabendazole, and ivermectin in order to evaluate the bioavailability and bioequivalence of a novel sheep anthelmintic formulation of oral suspension for sheep treatment containing triclabendazole 50 mg/mL and ivermectin 1 mg/mL compared to the reference product. In order to determine relative bioavailability of the test product with respect to the reference product the study was conducted on 36 clinically healthy sheep, following an unicentric, randomized, cross-over, two-sequence, two-treatment and 14-day wash-out study design. For the determination of triclabendazole sulphoxide and ivermectin sheep plasma concentrations, two rapid, selective high performance liquid chromatography coupled with mass spectrometry (LC-MS/MS) methods were developed and validated. The measured plasma concentrations of triclabendazole sulphoxide and ivermectin were used for the pharmacokinetic analysis and the determination of bioequivalence between the test product with regards to the reference product. The noncompartmental analysis of the pharmacokinetic data for both triclabendazole sulphoxide and ivermectin showed similarities between first-order kinetics of the test and reference product. The relevant pharmacokinetic parameters (Cmax, AUClast, AUCtot) were determined and the bioequivalence between the test and reference product could be concluded.

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The Complication Rates of Oral Anticoagulation Therapy in Deep Venous Thrombosis

DOI: 10.2478/amma-2019-0012

The objective of the current study is to evaluate the complication rates (embolic and hemorrhagic events) in deep venous thrombosis (DVT) patients on different types of oral anticoagulation therapy (OAC): direct oral anticoagulant therapy and vitamin K antagonist therapy.
Methods: A number of 62 DVT patients were included and divided in two groups, depending on the type of oral anticoagulation therapy administered. The first group was composed of patients treated with direct oral anticoagulant treatment (Dabigatran, Rivaroxaban) and the second group was composed of patients treated with vitamin K antagonist (Acenocumarol). General data, including BMI and comorbidities were noted. Embolic and hemorrhagic events were noticed.
Results: in the first group of patients (DOAC therapy), a number of 34 patients were included (14 of them with BMI higher than 25 kg/m2 and 14 with concomitant atrial fibrillation), while the second group comprised of 28 patients treated with VKA (21 of them with a high BMI and only 3 of them with atrial fibrillation). After a mean period of 36 months of anticoagulant therapy, complications were present in 17 patients, hematuria (8 episodes) and pulmonary embolism (4 cases) were the most frequent, with no difference regarding the treatment applied. Conclusion: No statistically significant difference was encountered regarding embolic and hemorrhagic event rates in our deep vein thrombosis patients.

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Development and Validation of an UV-Spectrophotometric Method for the Assay of Strontium Ranelate and HPLC Stability Testing from Bulk and Pharmaceutical Dosage Form

DOI: 10.2478/amma-2019-0014

Objective: The present work offers a fast, reliable and easy UV spectrophotometric method for the assay of strontium ranelate from bulk samples and pharmaceutical dosage form.
Methods: The proposed method uses 0.1% V/V trichloroacetic acid as dissolution medium for spectrophotometric analysis, by signal detection at 321 nm. The method was validated according to the currently in-force international guidelines for linearity, accuracy, precision, robustness, limit of detection and quantification.
Results: The method was found to be linear in the range of 5-100 µg mL-1 (R2 > 0.999). Method accuracy was found in-between 98.87-100.41%, showing good linear correlation as well (R2 = 0.9997). The concentrations for limit of detection and limit of quantitation were found 1.13 µg mL-1 and 3.77 µg mL-1, resp. The proposed method showed good intra- and interday precision, with low RSD values of 0.53-1.24% and 1.11%, resp.
Conclusions: Stability studies performed by both HPLC and UV spectrophotometric methods revealed that the active substance is highly susceptible to acidic hydrolysis, oxidation and exposure to high temperature.

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Analytical Quality by Design with the Lifecycle Approach: A Modern Epitome for Analytical Method Development

DOI: 10.2478/amma-2019-0010

Quality by Design is the methodical method to development concept that starts with the predefined objects. The method put emphasis on the process of development of a product, the control process, which is built on risk management and comprehensive knowledge of science. The concept of QbD applied to analytical method development is known now as AQbD (Analytical Quality by Design). Comprehension of the Analytical Target Profile (ATP) and the risk assessment for the variables that can have an impact on the productivity of the developed analytical method can be the main principles of the AQbD. Inside the method operable design region (MODR), the AQbD permits the movements of the analytical methods. This paper has been produced to discuss various views of analytical scientists, the comparison with conventional methods, and the phases of the analytical techniques.

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Development and Validation of an UHPLC Method for Ostarine Determination in Dietary Supplements

DOI: 10.2478/amma-2019-0008

Objective: The purpose of this study was to develop a low-cost, yet sensitive and precise UHPLC method for the quantitative determination of ostarine from dietary supplements (DS) for athletes. The analytical performance of the method was verified on a DS legally acquired from a specialized website for athletes. The uniformity of mass and content of the ostarine DS was also verified.
Methods: For the quantitative determination of ostarine a UHPLC method was developed and validated. The separation was performed using a reversed-phase C18 column, using a mixture of 75% methanol: 25% formic acid 0.1% in isocratic elution, at a flow rate of 0.5 ml/min. The uniformity of mass and content of DS was performed following the methodology described in the European Pharmacopoeia 7th Edition.
Results: The validated method was specific and linear on the concentration range of 1-25 µg/ml and was precise and accurate at all concentration levels, according to the official guidelines for validating analytical methods. An average mass of 510 mg content was obtained for the ostarine capsules, with an RSD of 2.41%. Regarding the uniformity of the content, an average of 4.65 mg ostarine/capsule was obtained with an RSD of 1.05%.
Conclusions: The developed UHPLC method was suitable, rapid, sensitive and allowed quantitative determination of active substance content in a DS with ostarine (92.91% ostarine/capsule from 5 mg ostarine/capsule declared by the manufacturer).

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Urinary Sodium/Potassium Ratio in Acute Kidney Injury Accurately Differentiates Prerenal Azotemia from Acute Tubular Necrosis

DOI: 10.2478/amma-2019-0011

Objective: To develop a more accurate, cost effective, non-invasive test to differentiate between pre-renal renal failure (PRA) and acute tubular necrosis (ATN) in acute kidney injury (AKI).
Methods: Urine sodium/potassium (Na/K) ratios were compared with fractional excretion of sodium (FeNa) and renal failure index (RFI) as well as other commonly used indices to differentiate patients with PRA from ATN. Patients with a rise in serum creatinine > 0.5 mg/d identified from medical records for a six- to eighteen-month period, were reviewed and categorized either as PRA or ATN based on presenting findings, course in hospital or renal biopsy. All patients had urinary sodium and potassium, creatinine, and serum creatinine done.
Results: The Na/K was < 1 in PRA and > 1 in ATN, correctly identifying all 42 cases of PRA and all 28 patients with ATN. The FeNa was >1 and misdiagnosed 9 of 42 patients with PRA and was >1 and correctly diagnosed all patients with ATN. The RFI was >1 and misdiagnosed 11 of 42 patients with PRA but was >1 and correctly diagnosed all patients with ATN. The BUN/creatinine ratio, urine sodium concentration and U/P creatinine ratio all had a very poor correlation with the correct diagnosis.
Conclusion: The Na/K ratio correctly diagnosed all 42 cases of PRA and all 28 cases of ATN. It is easy to do, is cost effective, non-invasive, and is useful for following patients with PRA to see if and when they develop ATN.

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A Prospective Study about the Influence of Selenium Based Supplements on the Autoimmune Process Evolution and the Health-Related Quality of Life in Patients with Chronic Autoimmune Thyroiditis

DOI: 10.2478/amma-2019-0009

Objective: The purpose of this study was to investigate the benefits of two different Selenium based supplements on patients with chronic autoimmune thyroiditis.
Methods: We conducted a prospective study on 50 patients with chronic autoimmune thyroiditis, who were divided into three different treatment groups, one group taking Selenium 100 µg, one Procor T (a combination of Selenium 100 µg and other elements like copper, Zinc and Q10 Conenzyme) and one control group taking Placebo pills. We measured on two follow up visits the antibody levels (anti-thyroidperoxidase- TPO Ab) and offered each patient a standardized questionnaire regarding the thyroid-related quality of life (THYPROro).
Results: At the 6 months follow up visit there was a statistically significant decrease in the antibody levels for each treatment group compared to the base levels. The THYPROro questionnaire scores showed an improvement in most aspects regarding the quality of life as well, but there was no significant difference between the placebo and the treated groups in the magnitude of this improvement.
Conclusions: Based on our results, we could not identify a certain benefit in improving quality of life with the supplementation of Selenium, as the improvements were at a similar level for the patients who took Placebo pills. Further studies with more patients, as well as taking the Selenium deficiency in consideration (by measuring the basal serum level of Selenium for each patient) would be required to find the target group of patients who could have most benefits of Selenium-based supplementation.

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New UHPLC Method for Cannabidiol Determination in Hard Capsules

DOI: 10.2478/amma-2019-0007

Objectives: The aim of the study was to propose a new UHPLC method for the determination of cannabidiol (CBD) from supplements and drugs available on the Romanian market. Materials and methods: The HPLC assay of CBD was achieved by using a Phenomenex Gemini NX-C18 column. The mobile phase consisted of 70% acetonitrile and 30% water. Elution was performed in isocratic mode and the detection was done at 208 nm. The method was tested on hard capsules containing 150 mg of CBD.
Results and discussions: The retention time of CBD was 2.8 minutes. Regression analysis showed good linearity over the 1-100 ug/ml concentration range. The lowest limit of quantification was established at 1 µg/ml. The method was developed by using reconstituted capsules. The substance proved low stability in solution at room temperature and stability at temperatures between 2-8ºC. The recovery of reconstituted samples was 96.77%. The commercial capsules had a very low content of 15-20% from declared content.
Conclusions: The proposed method can be used for CBD determination in different pharmaceutical formulations – hard and soft capsules with coconut oil as excipient.

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Dental Students’ Tobacco Smoking Habits, Second-hand Smoke Exposure, and Training in Cessation Counselling at the University of Medicine Pharmacy Sciences and Technology of Târgu Mureș

DOI: 10.2478/amma-2019-0006

Objectives: To describe tobacco smoking habits, attitudes, second-hand smoke exposure, and training in cessation counseling at the University of Medicine Pharmacy, Sciences and Technology of Târgu-Mureș (UMPSTTM), as baseline data for the first Romanian university to implement a Smoke-Free University Project.
Methods: A cross-sectional survey was administered in 2014 among dental students at UMPSTTM to explore their smoking habits, attitudes toward smoking and tobacco control policies, exposure to second-hand smoke, interest in quitting, and their knowledge about cessation counseling. We used core questions of the Global Health Professions Student Survey (GHPSS) and added specific items related to the Smoke-Free University Project. Data were analyzed by SPSS v22 software. We compared our results with those of the GHPSS Survey.
Results: 581 dental students, 73.1% of the target population (n=795), completed the questionnaire. 38.7% were current smokers. Approximately 1 in 5 (22.6%) current smokers admitted smoking inside university buildings, although 80.7% were aware of the smoking ban. 44.2% of current smokers plan to quit smoking. Nearly half of the students (48.9%) were exposed to second-hand smoke in their current homes, 78.1% in public places and 33.3% inside the university buildings. Only 21.0% of all participants received any formal training on how to help future patients quit.
Conclusions: Tobacco use prevalence was higher among future dentists than in the majority of respondents to the GHPSS. Changes in dental school education are needed to promote personal smoking cessation, as well as to educate dentists on how to support their future patients quitting.

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