Analysis of cationic liposomes by reversed-phase HPLC with evaporative light-scattering detection

Zhong, Zhenming; Ji, Qin; Zhang, J. Allen

doi:10.1016/j.jpba.2009.10.001

Cited by 48 publications

(23 citation statements)

References 31 publications

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“…Probe sonication of the liposomes for 10 min was shown to reduce the DMPC liposomes to ~90–100 nm with a PDI of 0.291 ± 0.011 ( Figure 6 ), and HPLC-ELSD analysis did not detect any degradation products resulting from sonication ( Figure 6 A). However, degradation products from DMPC were detected with HPLC after forced degradation ( Figure 6 B), as represented by a peak at 2.4 min equating to degradation products, as previously shown by Zhong and Zhang [ 29 ]. Zhong et al were able to show that dipalmitoyl phosphatidylcholine (DPPC) can degrade into palmitic acid and the lyso forms of DPPC.…”

Section: Resultssupporting

confidence: 74%

“…HPLC-ELSD settings were kept constant as outlined in Section 2.3 using the Phenomenex Luna 5 µm C8(2) 4.60 mm inner diameter and 150 mm length with 100 Ă pore size. The specificity of the method was tested by conducting forced degradation of a liposome formulation using 0.1N HCl at room temperature for four days, as previously described by Zhong and Zhang [ 29 ].…”

Section: Methodsmentioning

confidence: 99%

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Rapid Quantification and Validation of Lipid Concentrations within Liposomes

et al. 2016

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Quantification of the lipid content in liposomal adjuvants for subunit vaccine formulation is of extreme importance, since this concentration impacts both efficacy and stability. In this paper, we outline a high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) method that allows for the rapid and simultaneous quantification of lipid concentrations within liposomal systems prepared by three liposomal manufacturing techniques (lipid film hydration, high shear mixing, and microfluidics). The ELSD system was used to quantify four lipids: 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), cholesterol, dimethyldioctadecylammonium (DDA) bromide, and d-(+)-trehalose 6,6′-dibehenate (TDB). The developed method offers rapidity, high sensitivity, direct linearity, and a good consistency on the responses (R2 > 0.993 for the four lipids tested). The corresponding limit of detection (LOD) and limit of quantification (LOQ) were 0.11 and 0.36 mg/mL (DMPC), 0.02 and 0.80 mg/mL (cholesterol), 0.06 and 0.20 mg/mL (DDA), and 0.05 and 0.16 mg/mL (TDB), respectively. HPLC-ELSD was shown to be a rapid and effective method for the quantification of lipids within liposome formulations without the need for lipid extraction processes.

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Section: Resultssupporting

confidence: 74%

Section: Methodsmentioning

confidence: 99%

Rapid Quantification and Validation of Lipid Concentrations within Liposomes

et al. 2016

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“…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%

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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“…Thus, 1-acyl-2-lysoforms were analyzed. Consistent with previous reports describing reversed-phase HPLC methods for lipids (18), the HPLC separation was optimized using a C18 column with TFA or ammonium acetate in methanol. However, the mobile phase with TFA did not provide a reproducible retention time for DSPE-PEG, and thus the mobile phase with ammonium acetate was selected.…”

Section: Optimization Of the Separation Conditionsmentioning

confidence: 99%

“…Several HPLC-ELSD methods for the analysis of lipids or the hydrolysis products of phospholipids have been previously reported; however, the normal-phase separation was often used, which resulted in the large consumption of chloroform (14)(15)(16)(17). While the reversed-phase separation method for the analysis of lipids in cationic liposomes have been reported (18), the reversed-phase separation methods for the simultaneous analysis of PEG-conjugated lipid and the hydrolysis products in PEGylated liposomes have not been well studied. Thus, in this study, we attempted to develop a reversed-phase HPLC-ELSD system that could simultaneously analyze the lipid components and hydrolysis products in PEGylated liposomes.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Determination of Polyethylene Glycol-Conjugated Liposome Components by Using Reversed-Phase High-Performance Liquid Chromatography with UV and Evaporative Light Scattering Detection

2013

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Abstract. Liposomes incorporating polyethylene glycol (PEG)-conjugated lipids (PEGylated liposomes) have attracted attention as drug delivery carriers because they show good in vivo stability. The lipid component of PEGylated liposomal formulations needs to be quantified for quality control. In this study, a simple reversed-phase high-performance liquid chromatography (HPLC) method with an evaporative light-scattering detector (ELSD) was established for simultaneous determination of hydrogenated soy phosphatidylcholine, cholesterol, PEG-conjugated lipid, and hydrolysis products of phospholipid in PEGylated liposomal formulations. These lipids were separated using a C18 column with a gradient mobile phase consisting of ammonium acetate buffer and ammonium acetate in methanol at a flow rate of 1.0 ml/min. This method provided sufficient repeatability, linearity, and recovery rate for all lipids. However, the linearity and recovery rates of cholesterol achieved using a ultraviolet (UV) detector were better than those achieved using an ELSD. This validated method can be applied to assess the composition change during the preparation process of liposomes and to quantify lipid components and hydrolysis products contained in a commercially available liposomal formulation DOXIL®. Taken together, this reversed-phase HPLC-UV/ELSD method may be useful for the rapid or routine analysis of liposomal lipid components in process development and quality control.

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Analysis of cationic liposomes by reversed-phase HPLC with evaporative light-scattering detection

Cited by 48 publications

References 31 publications

Rapid Quantification and Validation of Lipid Concentrations within Liposomes

Rapid Quantification and Validation of Lipid Concentrations within Liposomes

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

Simultaneous Determination of Polyethylene Glycol-Conjugated Liposome Components by Using Reversed-Phase High-Performance Liquid Chromatography with UV and Evaporative Light Scattering Detection

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