HPLC assay for determination of amphotericin B in biological samples

Espada, Raquel; Josa, J.M.; Valdespina, Suriñe; Dea, M. A.; Ballesteros, M.; Alunda, José María; Torrado, J. J.

doi:10.1002/bmc.947

Cited by 33 publications

(41 citation statements)

References 18 publications

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“…The release kinetics of AmB was determined in two different release media: phosphate-buffered saline (PBS), pH 7.4, and sodium acetate buffer, pH 5.5, each containing 1% (vol/vol) Tween 80 (19,20). The amount of drug released was determined by high-pressure liquid chromatography (HPLC), using acetonitrile-acetic acid (10%)-water (41:43:16) as the mobile phase, a flow rate of 1.2 ml/min, a UV detector at 405 nm, and a C 18 column (4 mm ϫ 150-mm inner diameter) (21,22). The following equations were used to determine the drug encapsulation and loading efficiencies: drug encapsulation efficiency (%) ϭ (D 1 Ϫ D 2 )/D 1 ϫ 100 and drug loading efficiency (%) ϭ (D 1 Ϫ D 2 )/M ϫ 100, where D 1 is the amount of drug taken initially, D 2 is the amount of unentrapped drug, and M is the amount of dendrimeric carrier (23).…”

Section: Methodsmentioning

confidence: 99%

Surface-Engineered Dendrimeric Nanoconjugates for Macrophage-Targeted Delivery of Amphotericin B: Formulation Development andIn VitroandIn VivoEvaluation

Jain

Verma

Mishra

et al. 2015

Antimicrob Agents Chemother

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The present study aimed to develop an optimized dendrimeric delivery system for amphotericin B (AmB). Fifth-generation (5.0G) poly(propylene imine) (PPI) dendrimers were synthesized, conjugated with mannose, and characterized by use of various analytical techniques, including Fourier transform infrared spectroscopy (FTIR), 1 H nuclear magnetic resonance ( 1 H-NMR) spectroscopic analysis, and atomic force microscopy (AFM). Mannose-conjugated 5.0G PPI (MPPI) dendrimers were loaded with AmB and evaluated for drug loading efficiency, in vitro drug release profile, stability, hemolytic toxicity to human erythrocytes, cytotoxicity to and cell uptake by J774A.1 macrophage cells, antiparasitic activity against intracellular Leishmania donovani amastigotes, in vivo pharmacokinetic and biodistribution profiles, drug localization index, toxicity, and antileishmanial activity. AFM showed the nanometric size of the MPPI dendrimers, with a nearly globular architecture. The conjugate showed a good entrapment efficiency for AmB, along with pHsensitive drug release. Highly significant reductions in toxicity toward human erythrocytes and macrophage cells, without compromising the antiparasitic activity of AmB, were observed. The dendrimeric formulation of AmB showed a significant enhancement of the parasiticidal activity of AmB toward intramacrophagic L. donovani amastigotes. In the in vitro cell uptake studies, the formulation showed selectivity toward macrophages, with significant intracellular uptake. Further pharmacokinetic and organ distribution studies elucidated the controlled delivery behavior of the formulation. The drug localization index was found to increase significantly in macrophage-rich organs. In vivo studies showed a biocompatible behavior of MPPIA, with negligible toxicity even at higher doses, and promising antileishmanial activity. From the results, we concluded that surface-engineered dendrimers may serve as optimized delivery vehicles for AmB with enhanced activity and low or negligible toxicity. Nanomedicine has immense potential in drug delivery because of nanoscale size-derived chemical, physical, and biological properties. Dendritic polymers, including dendrigrafts, dendrimers, and dendrons, are providing new directions in nanomaterialbased drug delivery due to their nanostructure, monodispersity, versatility, well-defined molecular size, tailor-made surface groups, and chemical stability. The unique physicochemical and biological properties of dendrimers are due to their stepwise controlled synthesis, which is accomplished by sequential addition of layers/generations surrounding the core (1-4).Scientists are continuously investigating approaches for targeted delivery of antiparasitic drugs to macrophages because parasites reside and multiply within host macrophages in infectious diseases such as salmonellosis, leishmaniasis, histoplasmosis, listeriosis, cryptococcosis, candidiasis, AIDS, etc. Targeted delivery to macrophages in cases of infectious disease is expected to improve the therapeutic index by...

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Section: Methodsmentioning

confidence: 99%

Surface-Engineered Dendrimeric Nanoconjugates for Macrophage-Targeted Delivery of Amphotericin B: Formulation Development andIn VitroandIn VivoEvaluation

Jain

Verma

Mishra

et al. 2015

Antimicrob Agents Chemother

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“…The complex and AmBisome were administered in a 0.2 ml bolus by intravenous injection at an AMB equivalent dose of 1.25 mg/kg (body weight). AMB concentrations in plasma and organs were determined by HPLC by following an established method (21). Blood samples were obtained by cardiac puncture at 30 min and 2, 4, 6, and 24 h after drug injection.…”

Section: Methods (I) Preparation Of Amb-pga Complexmentioning

confidence: 99%

“…High-performance liquid chromatography (Jasco) analysis was conducted using a C 18 column (particle size, 5 m; length, 250 mm) (Fisher Scientific) by following an established analytical method (21). The mobile phase used was aceto-nitrile (52%), water (43.7%), and acetic acid (4.3%).…”

Section: Methods (I) Preparation Of Amb-pga Complexmentioning

confidence: 99%

Antileishmanial Activity, Uptake, and Biodistribution of an Amphotericin B and Poly(α-Glutamic Acid) Complex

Mohamed-Ahmed

Seifert

Yardley

et al. 2013

Antimicrob Agents Chemother

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bA noncovalent, water-soluble complex of amphotericin B (AMB) and poly(␣-glutamic acid) (PGA), with AMB loadings ranging from 25 to 55% (wt/wt) using PGA with a molecular weight range of 50,000 to 70,000, was prepared as a potential new treatment for visceral leishmaniasis (VL). The AMB-PGA complex was shown to be as active as Fungizone (AMB deoxycholate) against intracellular Leishmania donovani amastigotes in differentiated THP-1 cells. The in vitro uptake of the AMB-PGA complex by differentiated THP-1 cells was similar to that of Fungizone and higher than that of AmBisome (liposomal AMB). The AMB-PGA complex also displayed a dose-response profile similar to that of AmBisome in vivo in BALB/c mice against L. donovani, with 50% effective doses (ED 50 s) of 0.24 ؎ 0.03 mg/kg of body weight for the AMB-PGA complex and 0.24 ؎ 0.06 mg/kg for AmBisome. A biodistribution study with mice indicated that the AMB-PGA complex cleared more rapidly from plasma than AmBisome, with a comparable low level of distribution to the kidneys.

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“…Plasma was immediately harvested by centrifugation and analyzed for blood urea nitrogen (BUN) and plasma creatinine (PC) using commercially available kits (Leadman Biochemistry Co., Ltd, Beijing, China) (10,11).…”

Section: Nephrotoxicity Studymentioning

confidence: 99%

“…HPLC method was slightly modified for determination of AmB concentration (11,12). Briefly, 25 μl of the extracted sample was injected into a Phenomenex Luna C-18 column at 35°C.…”

Section: Hplc Analytical Assaymentioning

confidence: 99%

Development of Amphotericin B-Loaded Cubosomes Through the SolEmuls Technology for Enhancing the Oral Bioavailability

et al. 2012

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Abstract. The oral administration of amphotericin B (AmB) has the major drawback of poor bioavailability. The aim of this work was to evaluate the potential of AmB-loaded cubosomes as an oral formulation with improved bioavailability. This manuscript firstly developed AmB-loaded cubosomes by using the SolEmuls technology. The encapsulation efficiency, the in vitro release, and stability studies in simulated gastrointestinal fluid were used to evaluate AmB-loaded cubosomes. The acute nephrotoxicity, bioavailability, and tissue distribution study of AmB-loaded cubosomes were assayed upon oral administration to rats. SAXS and cryo-TEM exhibited AmB-loaded cubosomes as a bicontinuous cubic liquid crystalline phase with Pn3m geometry. The encapsulation efficiency and the results of in vitro release and stability studies in simulated gastrointestinal fluid further demonstrated that AmB was successfully encapsulated in cubosomes. AmB-loaded cubosomal formulation orally administrated in rats did not show nephrotoxicity and its relative bioavailability was approximately 285% as compared to Fungizone®. The AmB-loaded cubosomal formulation presented an effective potential approach for enhancing the oral bioavailability of AmB.

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HPLC assay for determination of amphotericin B in biological samples

Cited by 33 publications

References 18 publications

Surface-Engineered Dendrimeric Nanoconjugates for Macrophage-Targeted Delivery of Amphotericin B: Formulation Development andIn VitroandIn VivoEvaluation

Surface-Engineered Dendrimeric Nanoconjugates for Macrophage-Targeted Delivery of Amphotericin B: Formulation Development andIn VitroandIn VivoEvaluation

Antileishmanial Activity, Uptake, and Biodistribution of an Amphotericin B and Poly(α-Glutamic Acid) Complex

Development of Amphotericin B-Loaded Cubosomes Through the SolEmuls Technology for Enhancing the Oral Bioavailability

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