Determination of amphotericin B in PLA-PEG blend nanoparticles by HPLC-PDA

Abstract

In this work, we developed and validated an effective reversed-phase HPLC method with photodiode array (PDA) detection for the quantitative analysis of amphotericin B (AmB) in poly(lactide)-poly(ethylene glycol) (PLA-PEG) blend nanoparticles. Chromatographic runs were performed on a reverse phase C18 column using a mobile phase comprising a 9% acetic acid and acetonitrile mixture (40:60, v/v) under isocratic elution with a flow rate of 1 mL/min. AmB was detected at a wavelength of 408 nm. The validation process was performed considering the selectivity, linearity, precision, accuracy, robustness, limit of detection (LOD) and limit of quantitation (LOQ) of the method. A concentration range of 1-20 µg/mL was used to construct a linear calibration curve. The LOQ and LOD were 55 and 18 ng/mL, respectively. The mean recovery of AmB from the samples was 99.92% (relative standard deviation (RSD) = 0.34%, n=9), and the method was robust for changes in the flow rate of the mobile phase (maximum RSD=4.82%). The intra-and inter-assay coefficients of variation were less than 0.59%. The method was successfully used to determine the entrapment efficiency of AmB in PLA-PEG blend nanoparticles.Uniterms: Amphotericin B/determination. Amphotericin B/encapsulation. High performance liquid chromatography/reverse phase/quantitative analysis. Nanoparticles.Neste trabalho desenvolveu-se e validou-se um efetivo método por cromatografia líquida de alta eficiência (CLAE) em fase reversa com detecção por fotodiodos para a análise quantitativa de anfotericina B (AmB) em nanopartículas compostas por blendas de poli(ácido lático)-polietilenoglicol (PLA-PEG). Corridas cromatográficas foram realizadas sob coluna C18 de fase reversa com fase móvel consistindo de ácido acético 9% e acetonitrila (40:60, v/v), em eluição isocrática com fluxo de 1 mL/min. A AmB foi detectada no comprimento de onda de 408 nm. O processo de validação foi realizado considerando a seletividade, linearidade, precisão, exatidão, robustez, limite de detecção (LD) e limite de quantificação (LQ) do método. Uma faixa de concentração entre 1-20 µg/mL foi usada para obter a curva-padrão linear. Os valores de LD e LQ foram 55 e 18 ng/mL, respectivamente. A recuperação média da AmB a partir das amostras foi de 99,92% (desvio padrão relativo = 0,34%, n=9) e o método foi robusto, considerando alterações no fluxo da fase móvel (desvio padrão relativo máximo=4,82%). Os coeficientes de variação intra e inter dia foram inferiores a 0,59%. O método foi utilizado com sucesso para a determinação da eficiência de encapsulação da AmB em nanopartículas de PLA-PEG.Unitermos: Anfotericina B/determinação. Anfotericina B/encapsulação em nanopartículas. Cromatografia líquida de alta eficiência/fase reversa. Nanopartículas.

Ultradeformable Lipid Vesicles Localize Amphotericin B in the Dermis for the Treatment of Infectious Skin Diseases

“…The absorbance at 406 nm was recorded. EE was calculated using the following equation (eq )­ where A is the absorbance at 406 nm of the formulation after probe sonication but before centrifugation and B is the absorbance at 406 nm of the formulation after centrifugation.…”

“…1 The absorbance at 406 nm was recorded. EE was calculated using the following equation 46 (eq 4) Table 3. Selected Factors and Levels to Implement the Box-Behnken Design of Experiments for the Optimization of the AmB-TFs Formulation…”

“…Animals were arranged as follows: group 1 was treated with AmB-TFs (0.5 mg/mL, 20 mg of formulation/day, topically applied), group 2 was treated with Glucantime (50 mg/kg, 25 μL intralesionally injected), group 3 was the infected control group with no treatment, and group 4 consisted of untreated and uninfected animals (healthy control group). Lesion size was monitored at days 4,21,28,31,35,42,46,51, and 56 postinfection. Skin parasitism was quantified 10 days after the end of the treatment (at 56 days postinfection).…”

Nanoparticles as a Tool for Broadening Antifungal Activities

“…Different polymers have been used to make amphotericin B polymeric nanoparticles. Among the polymers, poly lactic-co-glycolic acid (PLGA) [207], poly (lactic acid) (PLA) [208], polyethylene glycol (PEG), polycaprolactone (PCL) [209], and chitosan [141] are the most common, being developed by different techniques [210,200]. The main advantage of using these systems is their biocompatibility and biodegradability [211].…”