HPLC analysis as a tool for assessing targeted liposome composition

Oswald, Mira; Platscher, Michael; Geißler, Simon; Goepferich, Achim

doi:10.1016/j.ijpharm.2015.11.014

Cited by 24 publications

(11 citation statements)

References 18 publications

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“…We note that ELSD is a powerful tool for detecting lipids that have no strong chromophore (49,50). Based on our experience, the sensitivity of ELSD is markedly superior, by at least ϳ100-fold, to that of a UV-visible absorption detector (e.g.…”

Section: Methodsmentioning

confidence: 92%

Mechanism for Remodeling of the Acyl Chain Composition of Cardiolipin Catalyzed by Saccharomyces cerevisiae Tafazzin

Abe

Hasegawa

Oku

et al. 2016

Journal of Biological Chemistry

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Remodeling of the acyl chains of cardiolipin (CL) is responsible for final molecular composition of mature CL after de novo CL synthesis in mitochondria. Yeast Saccharomyces cerevisiae undergoes tafazzin-mediated CL remodeling, in which tafazzin serves as a transacylase from phospholipids to monolyso-CL (MLCL). In light of the diversity of the acyl compositions of mature CL between different organisms, the mechanism underlying tafazzin-mediated transacylation remains to be elucidated. We investigated the mechanism responsible for transacylation using purified S. cerevisiae tafazzin with liposomes composed of various sets of acyl donors and acceptors. The results revealed that tafazzin efficiently catalyzes transacylation in liposomal membranes with highly ordered lipid bilayer structure. Tafazzin elicited unique acyl chain specificity against phosphatidylcholine (PC) as follows: linoleoyl (18:2) > oleoyl (18:1) ‫؍‬ palmitoleoyl (16:1) > > palmitoyl (16:0). In these reactions, tafazzin selectively removed the sn-2 acyl chain of PC and transferred it into the sn-1 and sn-2 positions of MLCL isomers at equivalent rates. We demonstrated for the first time that MLCL and dilyso-CL have inherent abilities to function as an acyl donor to monolyso-PC and acyl acceptor from PC, respectively. Furthermore, a Barth syndrome-associated tafazzin mutant (H77Q) was shown to completely lack the catalytic activity in our assay. It is difficult to reconcile the present results with the so-called thermodynamic remodeling hypothesis, which premises that tafazzin reacylates MLCL by unsaturated acyl chains only in disordered non-bilayer lipid domain. The acyl specificity of tafazzin may be one of the factors that determine the acyl composition of mature CL in S. cerevisiae mitochondria.

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Section: Methodsmentioning

confidence: 92%

Mechanism for Remodeling of the Acyl Chain Composition of Cardiolipin Catalyzed by Saccharomyces cerevisiae Tafazzin

Abe

Hasegawa

Oku

et al. 2016

Journal of Biological Chemistry

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“…S9) due to the dependence of the efficiency of prevalent light-scattering processes, namely Rayleigh scattering, Mie scattering, and reflection-refraction, on the average particle size [37,78,79]. As previously reported for the analysis of lipids using ELSD, a second-order polynomial (quadratic) of the following form was used as a fitting model: Y = a 0 X 2 + a 1 X + a 2 , where X represents the concentration in µg/mL, a 0 , a 1 , and a 2 are constants and Y is the peak area [21,38,[80][81][82]. This model produced R 2 (coefficient of determination) values > 0.9977 indicating that the fitting was statistically valid (Table 3).…”

Section: Calibrationmentioning

confidence: 99%

“…In a similar timeframe, colloidal stability was rapidly lost under accelerated thermal conditions (results not shown). As previously reported for liposomes, the hydrolysis of phospholipids by hydrolytic cleavage of the ester functionalities in presence of water generates lysophospholipids and free fatty acids that prevent them from acting as surfactants and strongly affects the physical stability of the O/W nanoemulsion by promoting fusion of the nanoparticles, mainly by Ostwald ripening [20,21,86]. Hydrolysis of PE, PC and SB was assessed by plotting lipid excipient concentration against storage time on a semi-logarithmic scale.…”

Section: Stability Of Lipid Excipients During Long-term Storage and Amentioning

confidence: 99%

“…From an analytical point of view, the quality of these pharmaceuticals must be assured through the development of appropriate methods to characterize and quantify their lipid components and degradation products [18,19]. For this purpose, high-performance liquid chromatography (HPLC) is the technique of choice because of its ability to separate lipids into different classes or species based on their alkyl chain lengths and/or head group polarity [20][21][22][23]. For instance, reversed-phase (RP)-HPLC has become the most popular method for separating and analyzing triglyceride species in oils and fats, because it operates on the principle of both chain length and degree of unsaturation of fatty acids, thus enabling separation of individual triglyceride molecules [24,25].…”

Section: Introductionmentioning

confidence: 99%

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Development and validation of a novel UPLC-ELSD method for the assessment of lipid composition of nanomedicine formulation

Varache

Ciancone

Couffin

2019

International Journal of Pharmaceutics

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This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Development and validation of a novel UPLC-ELSD method for the assessment of lipid composition of nanomedicine formulation.

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“…Although several HPLC methods have been reported to separate and quantify lipids using evaporative light scattering detection (ELSDs) (Jeschek et al, 2016;Oswald et al, 2016;Wöll et al, 2018). However, this detection method is not specific, and cannot distinguish liposomal formulation lipids from biological medium lipids which appears as a major limitation in using HPLC-ELSDs for our application.…”

Section: Quantitative Assessment By Hplc Equipped With Fluorescence Dmentioning

confidence: 99%

Qualitative and quantitative analysis of the uptake of lipoplexes by villous placenta explants

Alhareth

Valero

Mohamed

et al. 2019

International Journal of Pharmaceutics

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Controlled distribution of a drug by its association to a nanocarrier is a promising approach for the treatment of pregnancy disorders such as preeclampsia. For this application, tracking both the nanocarrier and the drug is necessary to ensure the safety of both the mother and the foetus. This study reports a method to visualize and quantify the uptake of liposomal formulations in placental tissue using florescent labelling and appropriate analytical tools. Lipoplexes were labelled with a fluorescent lipid, DOPE-NBD while the encapsulated siRNA was fluorescently labelled with rhodamine. Lipoplexes were incubated with villous placenta explants, explants were imaged with confocal microscopy, then DOPE-NBD was extracted from the explant and quantified by HPLC. Qualitative evaluation by confocal microscopy showed the presence of lipoplexes and siRNA into the outer layer of the placental explants, the syncytiotrophoblast. For quantitative evaluation, an HPLC method for the quantification of fluorescent lipid DOPE-NBD in placental tissue was developed and validated. The developed method was applied to quantify the DOPE-NBD uptake in the placental tissue. Increased amounts of DOPE-NBD were detected in placental explants when increasing the incubation concentration of lipoplexes. This study provides a method to evaluate the interactions between liposomal formulation and the placental barrier.

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HPLC analysis as a tool for assessing targeted liposome composition

Cited by 24 publications

References 18 publications

Mechanism for Remodeling of the Acyl Chain Composition of Cardiolipin Catalyzed by Saccharomyces cerevisiae Tafazzin

Mechanism for Remodeling of the Acyl Chain Composition of Cardiolipin Catalyzed by Saccharomyces cerevisiae Tafazzin

Development and validation of a novel UPLC-ELSD method for the assessment of lipid composition of nanomedicine formulation

Qualitative and quantitative analysis of the uptake of lipoplexes by villous placenta explants

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