Matrix Effects of the Hydroethanolic Extract of Calyces of Physalis peruviana L. on Rutin Pharmacokinetics in Wistar Rats Using Population Modeling.

Rutin is the rutinose conjugate of quercetin. It presents several biological activities and is the major flavonoid in the hydroalcoholic extract of the calyces of Physalis peruviana L. It also shows hypoglycemic activity after oral administration. The aim of this work was to study the matrix effects of the extract from P. peruviana calyces on the pharmacokinetics of rutin and its metabolites in Wistar rats, using non-compartmental and population pharmacokinetic analyses. A pharmacokinetic study was performed after intravenous and oral administration of different doses of pure rutin and the extract. In the non-compartmental analysis, it was found that rutin from the extract exhibited higher distribution and clearance, as well as an 11-fold increase in the bioavailability of its active metabolites. A population pharmacokinetic model was also carried out with two compartments, double absorption and linear elimination, in which the extract and the doses were the covariates involved. This model correctly described the differences observed between rutin as a pure compound and rutin from the extract, including the dose dependency.

Matrix effects of the hydroethanolic extract and the butanol fraction of calyces from Physalis peruviana L. on the biopharmaceutics classification of rutin.

OBJECTIVES: The Biopharmaceutics Classification System (BCS) categorizes active pharmaceutical ingredients according to their solubility and permeability properties, which are susceptible to matrix or formulation effects. The aim of this research was to evaluate the matrix effects of a hydroethanolic extract of calyces from Physalis peruviana L. (HEE) and its butanol fraction (BF), on the biopharmaceutics classification of their major compound, quercetin-3-O-rutinoside (rutin, RU). METHODS: Rutin was quantified by HPLC-UV, and Caco-2 cell monolayer transport studies were performed to obtain the apparent permeability values (Papp ). Aqueous solubility was determined at pH 6.8 and 7.4. KEY FINDINGS: The Papp values followed this order: BF > HEE > RU (1.77 ± 0.02 > 1.53 ± 0.07 > 0.90 ± 0.03 × 10-5  cm/s). The lowest solubility values followed this order: HEE > RU > BF (2.988 ± 0.07 > 0.205 ± 0.002 > 0.189 ± 0.005 mg/ml). CONCLUSIONS: According to these results, rutin could be classified as BCS classes III (high solubility/low permeability) and IV (low solubility/low permeability), depending on the plant matrix. Further work needs to be done in order to establish how apply the BCS for research and development of new botanical drugs or for bioequivalence purposes.

Pharmacokinetics of Botanical Drugs and Plant Extracts.

BACKGROUND: Botanical drugs contain plant extracts, which are complex mixtures of compounds. As with conventional drugs, it is necessary to validate their efficacy and safety through preclinical and clinical studies. However, pharmacokinetic studies for active constituents or characteristic markers in botanical drugs are rare. OBJECTIVE: The objective of this review was to investigate the global state of the art in pharmacokinetic studies of active ingredients present in plant extracts and botanical drugs. A review of pharmacokinetics studies of chemical constituents of plant extracts and botanical drugs was performed, with a total of 135 studies published between January 2004 and February 2015 available in recognized scientific databases. Botanical preparations were mainly found in the form of aqueous extracts of roots and rhizomes. The most widely studied species was Salvia miltiorrhiza Bunge, and the compound most frequently used as a pharmacokinetic marker was berberine. CONCLUSION: Most studies were performed using the Sprague Dawley rat model, and the preparations were mainly administered orally in a single dose. Quantification of plasma concentrations of pharmacokinetic markers was performed mainly by liquid-liquid extraction, followed by high performance liquid chromatography coupled to mass spectrometry detector. In conclusion, in recent years there has been an increasing interest among researchers worldwide in the study of pharmacokinetics of bioactive compounds in botanical drugs and plant extracts, especially those from the Traditional Chinese Medicine.

Validación de una metodología analítica USP por HPLC para la cuantificación de warfarina sódica en tabletas / Validation of an analytical HPLC method for the quantification of warfarin sodium in tablets

La metodología analítica para la cuantificación de warfarina sódica en tabletas de 5 mg fue estandarizada y validada mediante cromatografía líquida de alta eficiencia acoplada a detector de arreglo de diodo (HPLC-DAD). Se usó como fase móvil una mezcla compuesta por MeOH-AcOH-H2O (68:1:32), una columna cromatográfica C8e Phenomenex® de 150 x 4,6 mm y tamaño de partícula de 5 μm. Los resultados analíticos muestran que el método es preciso, exacto y selectivo para este fármaco. La curva de calibración se realizó en un intervalo del 80 al 120%, en una concentración nominal de 0,1mg/mL, demostrando ser lineal con un coeficiente de correlación a r²> 0,990; el porcentaje de recuperación fue de 98,1% ± 1,3, la precisión medida a través de la repetibilidad y la precisión intermedia fue adecuada (%RSD<2). En la prueba de disolución se encontró un porcentaje de 92,3% ± 3.0. Por lo tanto, la metodología desarrollada cumple con las especificaciones establecida por la USP 38/ NF33.

The analytic methodology for the quantification of sodium warfarin tablets of 5 mg was standardized and validated by High Efficiency Liquid Chromatography coupled to Diode Array Detector (HPLC-DAD). A mixture of MeOH-AcOH-H2O (68:1:32) was used like mobil phase, a chromatographic column C8e Phenomenex® of 150x4.6 mm and a particle size of 5 μm. The analytic results show that the method is precise, accurate and selective for this drug. The calibration curve was performed in the range of 80 to 120% at a nominal concentration of 0.1 mg/mL, demonstrating to be linear with a correlation coefficient at r²> 0.990; recovery rate was 98.1% ± 1.3 and the precision measured through repeatability and intermediate accuracy was adequate (% RSD <2). A percent of 92.3% ± 3.0 was found in dissolution test, thus, the methodology developed complies with the specifications established by USP 38/NF33.

Determinación de la mutación K76T de la proteína transportadora resistente a Cloroquina de Plasmodium falciparum (PFCRT) en individuos con malaria en una zona de transmisión moderada del Caribe colombiano / Determination of K76T mutation in the Plasmodium falciparum Chloroquine resistance transporter (PFCRT) in people with malaria in a zone of moderate transmission of the Colombian Caribbean

Introducción: La resistencia de Plasmodium falciparum a los antimalaricos, está relacionada con mutaciones puntuales en proteínas esenciales en la biología del parásito. La mutación K76T en la proteína transportadora resistente a Cloroquina (CRT) es un marcador molecular de resistencia a este fármaco. En Colombia se ha reportado una frecuencia del 100% a esta mutación. Objetivo: Determinar la frecuencia de la mutación K76T de CRT en Plasmodium falciparum, infectando sangre de individuos con malaria adquirida en una zona del Caribe Colombiano con transmisión moderada. Materiales y métodos: El ADN del parásito se extrajo de láminas diagnósticas usando Chelex-100. Un fragmento de 148 pb de Pfcrt se amplificó por PCR, la mutación K76T fue determinada por análisis de Polimorfismos en Longitud de Fragmentos de Restricción (RFLP) usando la endonucleasa Apo I, los productos fueron separados por electroforesis en geles de poliacrilamida (PAGE). Resultados: Se estudiaron 66 muestras de individuos con malaria adquirida en seis municipios del Magdalena. Se logró amplificación de Pfcrt en 56 y la mutación T76 fue detectada en 55 (98,2%). La presencia de una muestra con el alelo K76 silvestre indicaría la existencia de reversión de la mutación K76T en Colombia, este fenómeno se ha observado en algunos países del continente africano, donde se reportó recuperación de la sensibilidad a Cloroquina. Conclusión: Este estudio es un primer acercamiento hacia el conocimiento de la resistencia a los antimaláricos en zonas de transmisión moderada en Colombia.

Introduction: The antimalarial drugs resistance from Plasmodium falciparum is related to point mutations in the essential proteins to the parasite biology. The K76T mutation in the Plasmodium falciparum Chloroquine resistance transporter (PFCRT) is a molecular marker resistance to Chloroquine. In Colombia the frequency of this mutation is 100%. Objective: To determine the frequency of K76T mutation in the Plasmodium falciparum CRT, infecting blood from people with malaria in the Colombia Caribbean zone with moderate transmission. Materials and methods: The DNA of the parasite was extracted from diagnostic slide using Chelex-100. A fragment of 148 pb of Pfcrt was amplified by PCR, the K76T mutation was evaluated by Restriction Fragment Length Polymorphism (RFLP) using endonuclease Apo I, the products were separated by polyacrylamide gel electrophoresis (PAGE). Results: Sixty six samples from people with malaria of six municipalities of Magdalena were studied. Fifty six DNA samples amplified of Pfcrt gene, T76 mutation was detected in 55 (98.2%). Occurrence of one sample carried the wild allele K76 would indicated the reversion of K76T mutation in Colombia, this phenomenon has been observed in some countries of the African continent, where recovery of sensitivity to Chloroquine was reported. Conclusions: This study is the first approach towards the knowledge of the antimalarials resistance in zones with moderate transmission in Colombia.