Effect of water and protic solvents on polysaccharide-based column efficiency.

In the present study, polysaccharide-based columns were used to evaluate the efficiency of columns in response to the introduction of water and protic solvents (methanol and ethanol) into the mobile phase, replacing acetonitrile. While increasing water content frequently enhances enantiomer resolution, the inclusion of water, particularly when combined with methanol and ethanol in the mobile phase, has an adverse impact on mass transfer, thus influencing the column plate height. These effects are more pronounced with ethanol, and in many cases, van Deemter plots exhibit the absence of a minimum point optimal in the explored range. Consequently, acetonitrile and its water mixtures are the preferred choices to mitigate these effects for situations in which the chiral column is operated at a relatively high flow rate (> 1 mL/min in a 4.6 mm column).

Blood Bag Plasticizers Influence Red Blood Cell Vesiculation Rate without Altering the Lipid Composition of the Vesicles.

BACKGROUND: Polyvinyl chloride (PVC) plasticized with di(2-ethylhexyl) phthalate (DEHP) is commonly used for blood collection and storage. DEHP has protective effects on RBC membranes, but is also a toxin. METHODS: A paired study was conducted to investigate the influence of DEHP and two alternative plasticizers, 1,2-cyclohexane-dicarboxylic acid diisononyl ester (DINCH) and n-butyryl-tri-n-hexyl citrate (BTHC), on the preservation of RBCs stored for 42 days in PVC pediatric bags. The RBC membrane was evaluated for supernatant hemoglobin (Hb), release of extracellular microvesicles (EVs), osmotic fragility, deformability, and lipid composition. RESULTS: In BTHC-plasticized bags, the supernatant Hb increase during storage was 2 times greater than in DINCH- and DEHP-plasticized bags. By day 21, EV concentrations had doubled from day-5 levels in DINCH- and DEHP-, and trebled in BTHC-plasticized bags. RBC mean cell volumes were greater in BTHC- than in DINCH- or DEHP-plasticized bags (p < 0.001). Osmotic fragility differed significantly among plasticizers (p < 0.01). After day 21, RBC deformability decreased in all, but to a greater extent in the bags with BTHC. Phospholipid composition of RBCs and EVs was not different among plasticizers. CONCLUSION: Membrane stabilization capacity differed among the plasticizers. RBC in BTHC bags stored more poorly, while DEHP and DINCH bags offered better protection against vesiculation, osmotic stress, and Hb loss.

A Phase I clinical trial of lodenafil carbonate, a new phosphodiesterase Type 5 (PDE5) inhibitor, in healthy male volunteers.

UNLABELLED: Lodenafil carbonate is a new phosphodiesterase Type 5 (PDE5) inhibitor used in treatment of erectile dysfunction. OBJECTIVE: The present study was conducted to evaluate the safety, tolerability, and pharmacokinetics of lodenafil carbonate after administering ascending (1 - 100 mg) single oral doses to healthy male volunteers (n = 33). METHODS: The study was an open label, dose-escalation, Phase I clinical trial involving the administration of single oral doses of lodenafil carbonate. Lodenafil carbonate was administered sequentially, escalating in single doses of 1 mg - 100 mg with a washout period of at least 1 week between each dose. The progression to the next dose was allowed after clinical and laboratory exams, Ambulatory Monitoring of Arterial Pressure (AMAP) without relevant clinical modifications and adverse events without clinical relevancy. Blood samples were collected at pre-dose, 0.25, 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 10, 12, 14, 16, 20 and 24 h post-dosing. Plasma samples for measurement of lodenafil carbonate and lodenafil were analyzed by liquid chromatography coupled to tandem mass spectrometry. RESULTS: No serious adverse events were observed, and none of the subjects discontinued the study due to intolerance. The AMAP measurements, clinical and laboratory exams and ECG revealed no significant changes even at higher doses. Lodenafil carbonate was not detected in any samples, indicating that it acts as a prodrug. The mean lodenafil pharmacokinetic parameters for tmax and t1/2 were 1.6 ( ± 0.4) h and 3.3 ( ± 1.1) h, respectively. This study demonstrated that lodenafil carbonate was well tolerated and showed a good safety profile in healthy male volunteers.

Efficiency of the same neat silica column in hydrophilic interaction chromatography and per aqueous liquid chromatography.

The dependencies on the mobile phase flow velocity of the efficiency of a column packed with shell particles of neat porous silica (Halo) was measured under two different sets of experimental conditions. These conditions corresponded to the retention mechanisms of per aqueous liquid chromatography (PALC) at low acetonitrile concentrations and of hydrophilic interaction chromatography (HILIC) at high acetonitrile concentrations. The results are compared. Small amounts of a diluted solution of caffeine were injected in order to record the chromatograms under strictly linear conditions. These efficiencies were measured in both water-rich (PALC retention mechanism) and acetonitrile-rich (HILIC mechanism) mobile phases for the same retention factors, between 0.25 and 2.5. The mobile phases were mixtures of acetonitrile and water containing neither supporting salt nor buffer component. At low retention factors, the efficiency of caffeine is better in the PALC than in the HILIC mode. For k(')=0.5, the minimum reduced height equivalent to a theoretical plate (HETP) is close to 2.5 in PALC while it exceeds 5 in HILIC. The converse is true for high retention factors. For k(')>2.5, the HETP is lower in HILIC than in PALC, because the major contribution to band broadening and peak tailing in this latter mode originates from the heterogeneous thermodynamics of retention and eventually restricts column performance in PALC. Most interestingly, the reduced HETP measured in HILIC for caffeine never falls below 4. This suggests that the mass transfer of caffeine between the multilayer adsorbed phase (due to the interactions of the strong solvent and the silanol groups) and the acetonitrile-rich bulk eluent is slow.

Evaluation of a new polymeric stationary phase with reversed-phase properties for high temperature liquid chromatography.

The performance of a polymeric stationary phase with reversed-phase properties (ET-RP1) was evaluated for LC separations at elevated temperature. The most significant observation was that the reduced plate height (h) decreased from 3.4 at 25 degrees C (optimal flow 0.5 mL/min) to 2.4 at 150 degrees C (optimal flow 2.5 mL/min) which is comparable to the efficiency obtained with silica-based reversed-phase columns of 4.6mm ID operated at 0.8 mL/min. The phase showed no deterioration after long use at 150 degrees C within the pH range 1-9. Catalytic activity originating from the stationary phase material, e.g. as experienced on zirconium columns operated at elevated temperature, was absent. The performance of ET-RP1 is illustrated with the analysis of some pharmaceutical samples by LC and LC-MS. Operation at elevated temperature also allows to reduce the amount of organic modifier or to replace acetonitrile and methanol by the biodegradable ethanol.

The acetonitrile shortage: is reversed HILIC with water an alternative for the analysis of highly polar ionizable solutes?

In hydrophilic interaction chromatography (HILIC), best results are obtained with high concentrations of ACN. In the framework of green chromatography and the present shortage and very high price of this hazardous solvent, reversing the stationary phase to apolar and the mobile phase to aqueous can be of interest for several applications. The features of the aqueous RP technique called per aqueous LC (PALC) are illustrated with the analysis of catecholamines, nucleobases, acids, and amino acids. The ca. three-fold higher viscosity of water compared to ACN has consequences on the shape of the Van Deemter plot. For dopamine (N = 26.450 on a 25 cm x 4.6 mm id, 5 microm bare silica column), a reduced plate height of 1.9 at an u(opt) of 0.3 mm/s was calculated. The plate number, however, strongly depends on pH and ionic strength. As in RP separations, retention is shortened by adding an organic modifier. In the framework of green chromatography, the biodegradable ethanol was used. On the other hand, retention increased by lengthening the carbon chain of ion-pair reagents supporting the RP mechanism as well.

Distribution of quinic acid derivatives and other phenolic compounds in Brazilian propolis.

The quinic acid derivatives (including 4-feruoyl quinic and 5-ferruoyl quinic acids characterized for first time in propolis samples) and other phenolic compounds were quantified in thirteen Brazilian propolis samples by HPLC analysis. For chemometrical analysis, the distribution of quinic acid derivatives and other phenolic compounds were considered. The results suggest that the Brazilian propolis with floral origin from Citrus sp. have the highest concentration of the quinic acid derivatives (between 11.0 to 58.4 mg/mg of the dried crude hydroalcoholic extract) and therefore would probably show a more effective hepatoprotective activity.

Chemical composition of Tipuana tipu, a source for tropical honey bee products.

Tipuana tipu (Benth.) Kuntze is a tree from the leguminosae family (Papilionoideae) indigenous in Argentina and extensively used in urbanism, mainly in Southern Brazil. The epicuticular waxes of leaves and branch, and flower surface were studied by high temperature high resolution gas chromatography. Several compounds were characterized, among which the aliphatic alcohols were predominant in branch, leaves and receptacle. Alkanes were predominant only in the petals and the aliphatic acids were predominant in stamen. In branches and leaf epicuticular surfaces, six long chain wax esters series were characterized, as well as lupeol and b-amyrin hexadecanoates.