Factors contributing to variability in metformin concentration in polycystic ovary syndrome.

AIM: To investigate the factors affecting metformin concentrations after chronic administration in patients with polycystic ovary syndrome (PCOS), focusing on the pharmacokinetic variability and its implications for personalized therapy. METHODS: This study enrolled 53 PCOS patients undergoing long-term metformin treatment at the Clinic for Gynecology and Obstetrics in Nis, Serbia, from February to December 2019. Pharmacokinetic parameters were measured from blood samples, and metformin concentrations were determined with validated analytical techniques. RESULTS: There was a significant variability in metformin concentrations among PCOS patients, with body mass index (BMI) identified as a major influencing factor. Higher BMI was associated with lower plasma metformin levels, a finding suggesting an altered pharmacokinetic profile in obese patients. CONCLUSIONS: This study highlights the critical role of BMI in influencing metformin pharmacokinetics in PCOS patients and underscores the need for personalized treatment strategies in patients with PCOS.

Sample pretreatment and HPLC determination of antiplatelet drug ticagrelor in blood plasma from patients with acute coronary syndromes.

A simple HPLC method was developed for the determination of antiplatelet drug ticagrelor (TCG) in blood. Sample preparation and extraction conditions were investigated and optimized. The preparation of blood plasma was investigated by protein precipitation using perchloric acid, methanol, acetonitrile (ACN), and trifluoroacetic acid. Protein precipitation using ACN was found to be the most suitable. Chromatographic separation of TCG was performed on a C18 column with a mobile phase consisting of ACN and 15 mM ammonium acetate buffered at pH 8.0. The method was applied to determine TCG in blood plasma of patients who had a heart attack. Blood samples were collected 1.5 h after the administration of the initial loading dose of the antiplatelet drug. The average concentration of TCG was found to be 0.97 ± 0.53 µg/ml. The developed method proved to be very selective, without interferences from other endogenous substances and the influences of possible coadministered drugs. The limits of detection and quantification estimated by the signal-to-noise ratio in real samples were 0.24 and 0.4 µg/ml, respectively. The developed method is simple and can be easily applied in clinics and emergency cardiac situations after the initial loading dose of TCG in the first few hours of a heart attack.

In vitro effect of beer, red and white wine on the morphology and surface roughness of human enamel.

BACKGROUND: Beer, red and white wine are acidic drinks whose frequent consumption can increase the risk of dental erosion. OBJECTIVES: To establish the effect of beer, red and white wine on the morphology and surface roughness (SR) of human enamel using different exposure times in a cyclic deand remineralization model in vitro. MATERIAL AND METHODS: The experiment included 33 surgically extracted impacted human third molars from patients aged 18-25 years. Enamel samples obtained by cutting crowns (n = 132) were submitted to alternate cycles of demineralization in (1) beer, (2) red wine, (3) white wine, (PC) positive control (orange juice), and remineralization in artificial saliva, which also represented a medium for negative control (NC). The experiment included cycles with different exposure times in alcoholic beverages and orange juice of 15, 30 and 60 min. Thus, 12 groups were formed (for each drink and each exposure time) containing 10 samples each, while the NC group consisted of 12 samples. Experiments were repeated 3x/day for 10 days. Enamel surface alterations were determined by stylus profilometry (average surface roughness (Ra)) and scanning electron microscopy (SEM). The Shapiro-Wilk test, independent samples Kruskal-Wallis test and multiple comparisons (all pairwise) were performed. RESULTS: With increasing exposure time, there was a positive correlation with Ra for white wineand orange juice-immersed samples (60 min compared to 15 min), which was also observed using SEM. There was no significant difference in the Ra between the other experimental samples for the same exposure time. CONCLUSIONS: This study confirms a certain erosive potential of beer, red and white wine, and a significant relationship with pH, titratable acidity (TA) and SR, but not with the exposure time for all tested alcoholic beverages. Moreover, differences among the ultrastructural patterns caused by alcoholic beverages over the enamel surface were observed.

Caffeine in Hair Care and Anticellulite Cosmetics: Sample Preparation, Solid-Phase Extraction, and HPLC Determination.

Caffeine is extensively used in cellulite and hair growth cosmetic products. Regulations in the field of cosmetics require manufacturers to list caffeine in the ingredient list on product labels, but its exact content in these products is not declared. On the other hand, daily exposure to caffeine from all sources may approach health reference values. For that reason, it is important to know the exact caffeine content in products for skin and hair care. Cosmetics are often viscous or semisolid products of very complex chemical composition. To analyze caffeine in these complex sample matrices by liquid chromatographic methods, an extraction step is often necessary. This article presents the applicability of the solid-phase extraction (SPE) procedure for the caffeine extraction and high-performance liquid chromatography (HPLC) determination in anticellulite gels, shampoos, and hair balsam. The samples of gels were centrifuged after ammonia addition to precipitate carbomer. In cellulite reduction, gel caffeine content was found to be in the range of 0.7-1.7%, whereas in the hair-care products, it was about 1.0%.

Ultrastructural changes in the cemento-enamel junction caused by acidic beverages: An in vitro study.

The present in vitro study was aimed at evaluating the morphological changes in the cemento-enamel junction (CEJ) after exposure to acidic beverages using the scanning electron microscopy (SEM). The initial pH and titratable acidity (TA) was analyzed from follow groups: (I) Coca cola, (II) orange juice, (III) Cedevita, (IV) Red Bull, (V) Somersby cider, and (VI) white wine. The CEJ samples (n = 64), obtained from unerupted third molars, were allocated to one control (artificial saliva, n = 16) and six experimental groups (n = 8). The experimental samples were immersed in beverages (50 ml) for 15 min, three times daily, 10 days, and in artificial saliva between immersions. SEM analysis was performed in a blind manner, according to scoring scale. One-way ANOVA and Tukey's post hoc tests, as well as Kruskal-Wallis and Mann-Whitney U test used for statistical analysis. The pH values of the acidic beverages ranged from 2.65 (Coca cola) to 3.73 (orange juice), and TA ranged from 1.90 ml (Coca cola) to 5.70 ml (orange juice) of NaOH to reach pH 7.0. The SEM analysis indicated statistically significant differences between the control samples and those immersed in acidic beverages. The Groups IV, I, and II, showed the highest CEJ damage grade while those of the Group VI were the lowest. All the tested acidic beverages caused morphological changes in the CEJ with a smaller or larger exposure of dentine surface, and were not always related to the pH or TA of acidic beverages.

Maternal age has more pronounced effect on breast milk retinol and ß-carotene content than maternal dietary pattern.

OBJECTIVE: The effects on breast milk composition of advanced maternal age and maternal dietary habits during pregnancy and lactation have not, to our knowledge, been investigated in southeastern Europe and the Balkans. The aim of this study was to compare the content of retinol and ß-carotene in colostrum and mature milk samples obtained from different maternal age (MA) groups and to assess the potential relationship with maternal and demographic characteristics, dietary patterns, and lifestyle habits during pregnancy and lactation. METHODS: Forty-three nursing mothers were divided in two groups according to MA: ≥35 y of age (n = 22) and <35 y of age (n = 21). Total lipid concentrations were determined by gravimetric method, whereas retinol and ß-carotene contents were assessed by high-performance liquid chromatography method. Dietary patterns during pregnancy and lactation were assessed using food frequency questionnaires, and principal component analysis (PCA) statistical analysis was performed. RESULTS: Except for retinol levels in mature milk, significantly higher levels of total fats, retinol, and ß-carotene were found in the older group. Results of PCA analysis showed that MA was strongly correlated with fat content, retinol, and ß-carotene levels in colostrum samples, whereas in mature milk samples MA was highly correlated with fat content and moderately with ß-carotene. In terms of dietary patterns, retinol contents in milk samples from both groups were weakly to moderately correlated with consumption frequency of eggs, meat, milk and dairy products, whereas ß-carotene contents were weakly to strongly associated with consumption frequency of fruits and vegetables. CONCLUSIONS: PCA analysis used in the study clearly confirmed that MA, total lipids, retinol, and ß-carotene levels might serve as a good criterion for delimitation of breast milk samples collected in different stages of lactation from mothers of various ages. The present findings could represent key basis for further investigations.

Physicochemical and biochemical parameters in milk of Serbian breastfeeding women.

BACKGROUND/AIM: This study was undertaken to determine the changes and relationships between some important milk constituents as well as physical, rheological, and biochemical parameters of milk obtained from Serbian breastfeeding mothers. MATERIALS AND METHODS: Physicochemical and biochemical parameters and the concentrations of vitamins, uric acid, and minerals were determined during the three periods of lactation covering colostrum, transitional, and mature milk collected from 67 mothers who had a term-pregnancy. RESULTS: Large interindividual variations regarding many parameters were found between mothers at the same period of lactation, but the average values were mostly in the expected and recommended ranges. For some parameters, our values are quite different in relation to the milk of women from other countries or data reported by other authors. CONCLUSION: Differences in vitamin and mineral contents and physicochemical and rheological characteristics of milk obtained by Serbian breastfeeding mothers compared to that of mothers from other parts of the world have been found. This paper presents the measured data of some physical parameters of human milk about which there is little information in the literature.

Erosive Effect of Different Soft Drinks on Enamel Surface in vitro: Application of Stylus Profilometry.

OBJECTIVE: To assess the erosive potential of various soft drinks by measuring initial pH and titratable acidity (TA) and to evaluate enamel surface roughness using different exposure times. MATERIALS AND METHODS: The initial pH of the soft drinks (group 1: Coca-Cola; group 2: orange juice; group 3: Cedevita; group 4: Guarana, and group 5: strawberry yoghurt) was measured using a pH meter, and TA was measured by titration with NaOH. Enamel samples (n = 96), cut from unerupted human third molars, were randomly assigned to 6 groups: experimental (groups 1-5) and control (filtered saliva). The samples were exposed to 50 ml of soft drinks for 15, 30 and 60 min, 3 times daily, during 10 days. Between immersions, the samples were kept in filtered saliva. Enamel surface roughness was measured by diamond stylus profilometer using the following roughness parameters: Ra, Rq, Rz, and Ry. Data were analyzed by one-way ANOVA, Tukey's post hoc and Student-Newman-Keuls post hoc tests. RESULTS: The pH values of the soft drinks ranged from 2.52 (Guarana) to 4.21 (strawberry yoghurt). Orange juice had the highest TA, requiring 5.70 ml of NaOH to reach pH 7.0, whereas Coca-Cola required only 1.87 ml. Roughness parameters indicated that Coca-Cola had the strongest erosion potential during the 15 min of exposure, while Coca-Cola and orange juice were similar during 30- and 60-min exposures. There were no significant differences related to all exposure times between Guarana and Cedevita. Strawberry yoghurt did not erode the enamel surface regardless of the exposure time. CONCLUSION: All of the tested soft drinks except yoghurt were erosive. Erosion of the enamel surfaces exposed to Coca-Cola, orange juice, Cedevita, and Guarana was directly proportional to the exposure time.

HPLC method development for determination of doxycycline in human seminal fluid.

The present paper reports the development and validation of an analytical method for doxycycline quantification in human seminal fluid by HPLC with UV detection. The separation of doxycycline was achieved at 40°C on a reversed-phase C18 column using isocratic elution. The mobile phase consisted of acetonitrile (A) and water buffered at pH 2.5 with a concentrated orthophosphoric acid (B) in the volume ratio of 20:80 (v/v), respectively. The detection was performed at 350nm. As an internal standard (IS), tetracycline was used. The proposed method involves the extraction of doxycycline from seminal fluid based on acidic precipitation of the proteins using perchloric acid. The method showed good intra- and inter-day precisions (RSD<7.0%), good accuracy (recovery for doxycycline>80%), and high correlation coefficient (r=0.998) for standards subjected to the entire procedure. The detection and quantification limits were 0.087µg/ml and 0.264µg/ml. The developed method was used to analyze doxycycline in the seminal fluids obtained from male subjects who were treated with doxycycline-hyclate. The mean doxycycline concentrations of 0.89±0.07µg/ml and 0.45±0.26µg/ml were detected in seminal fluid after 6h and 12h, respectively. This is the first study reporting extraction and HPLC determination of doxycycline in this complex sample and can be very useful in support of clinical and pharmacokinetic studies on this antibiotic.

Determination of ibuprofen in combined dosage forms and cream by direct UV spectrophotometry after solid-phase extraction.

Solid-phase extraction method followed by direct UV spectrophotometry at 264 nm was developed and applied for the selective ibuprofen determination in two-component formulation of ibuprofen and pseudoephedrine-HCl, combined powder which contains ibuprofen in the form of salt with L-arginine and 10% ibuprofen cream. Procedures for ibuprofen determination in complex pharmaceutical preparations by direct UV spectrophotometry lack selectivity because of interferences of other active substances and fat components. A limited number of spectrophotometric methods applicable to these samples are based on derivative (first and second-order) UV spectroscopy. Common HPLC procedures are more selective but more expensive and for creams also require some type of extraction because the large amount of oily excipients would clog up the column. The proposed solid-phase extraction method proved to be suitable for analysis of ibuprofen in combined tablets, powders and creams by direct UV spectrophotometry. Also the method provides an effective clean-up of the cream and allows ibuprofen determination by HPLC analysis. For the extraction three different commercial sorbents were tested: anion exchange Oasis MAX, hydrophilic-lipophilic balanced Oasis HLB and reverse-phase Chromabond C18ec. The optimization of the SPE method was first done on standard ibuprofen solutions and then the suitability of the method was checked on solutions of commercial pharmaceutical samples. The method yields good results for all three types of commercial preparations on the anion-exchange Oasis MAX cartridges, with recoveries of 90-100.2%. The interferences in UV analysis were not registered and good precision (RSD < 6%) was obtained. The present method has been verified as accurate as the reference HPLC with the great advantage of less expensive instrumentation. For this reason, the method would be suitable for a routine and rapid drug quality control.

RP-HPLC assay of doxycycline in human saliva and gingival crevicular fluid in patients with chronic periodontal disease.

A reversed-phased HPLC method with fluorescence detection was optimized and validated for determination of DOXY in human saliva and gingival crevicular fluid (GCF) with tetracycline as internal standard. Single step extraction with acetonitrile for both types of samples was performed. The separation was achieved at Zorbax Extend-C18 analytical column at 30°C. Mobile phase was consisted of an aqueous phase containing magnesium acetate, ammonium acetate, Na2EDTA, triethyl-ammonium acetate buffered to pH 7.5 with ammonium hydroxide solution and acetonitrile. The volume ratio of the buffered water mixture of salts and acetonitrile was 86:14. Fluorescence detector was set at λex=380 nm and λem=520 nm. Under the optimized experimental conditions, good linearity was found in the range of 5.0-250.0 ng/mL for GCF with LOD of 1.63 ng/mL and LOQ of 4.93 ng/mL and 20.0-500.0 ng/mL for saliva with LOD of 6.36 ng/mL and LOQ of 19.28 ng/mL. This method was successfully applied for determination of DOXY in saliva and GCF obtained from patients with chronic periodontal disease.

Determination of streptomycin in a pharmaceutical sample based on its degradation by hydrogen peroxide in the presence of copper(II).

The kinetics of streptomycin degradation by hydrogen peroxide at pH 7.4 was investigated. The reaction was catalyzed by traces of Cu(II) ions, and it was followed spectrophotometrically at 322 nm by applying the initial-rate method. The kinetic parameters of the reaction are reported, and a rate equation is suggested. From the dependence of the relative rate constants on the temperature, the activation energy was calculated to be 57.5 kJ/mol. Based on this reaction, a kinetic method for streptomycin determination is proposed. The calibration graph is linear in the ranges 1.94 - 15.48 microg/cm(3) and 0.15 - 1.94 microg/cm(3). The influence of foreign ions and molecules on the accuracy of the determinations was investigated. The proposed kinetic method has high selectivity and good sensitivity, and enabled to determine streptomycin in pharmaceutical samples.