Development of a size exclusion chromatography cartridge-based analytical method for determination of free drug in nano-liposomal oncology drug formulations.

IF 2.6 3区 化学 Q2 CHEMISTRY, ANALYTICAL
Wei Zhang, Mark Paciolla, Lijun Duan, Elise Bradley, Aastha Chadha, Bhavesh Barot, Kaylee Worrell
{"title":"Development of a size exclusion chromatography cartridge-based analytical method for determination of free drug in nano-liposomal oncology drug formulations.","authors":"Wei Zhang, Mark Paciolla, Lijun Duan, Elise Bradley, Aastha Chadha, Bhavesh Barot, Kaylee Worrell","doi":"10.1039/d4ay02124j","DOIUrl":null,"url":null,"abstract":"<p><p>A reliable and stability-indicating size exclusion chromatography (SEC) cartridge-based free drug testing method was developed for active loading nanoliposome formulations through a systematic development approach. The SEC spin cartridge columns (7K MWCO, 2 mL) were used for developing the liposome free drug testing procedure. The SEC cartridge column retention capacity for a Mirati drug was determined (445 μg per cartridge). SEC testing conditions were studied to achieve a good separation between liposomes and the free drug, including cartridge conditioning, buffer wash steps for liposomal drug elution, organic media wash steps for free drug elution, and sample size effect. Qualification of this newly developed SEC cartridge method has demonstrated its specificity/selectivity without interference and excellent detection linearity (correlation coefficient (<i>R</i>) > 0.999) over a study concentration range (1.0 to 46.9 μg mL<sup>-1</sup>), sufficient LOQ (0.52 μg mL<sup>-1</sup> equivalent to 1.7% of free drug in a liposome formulation at 2.5 mg mL<sup>-1</sup>), acceptable accuracy/recovery of 81 to 89% for free drug in spiked samples at 4.5%, 9%, and 18% levels (sample loading by a regular pipet) and 88 to 97% at 10% spike level (sample loading by a positive displacement pipet), good method precision (RSD (<i>n</i> = 6) of 4% NMT) for free drug determination, and 3 days solution stability for both standard and sample solutions (2-8 °C). In comparative sample testing, the SEC free drug test results were in good agreement with the solid phase extraction (SPE) test results for active loading formulations. The new method's stability-indicating ability has been proved through monitoring free drug in a liposome sample stored at -20 °C and -80 °C.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":" ","pages":""},"PeriodicalIF":2.6000,"publicationDate":"2025-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Analytical Methods","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1039/d4ay02124j","RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
引用次数: 0

Abstract

A reliable and stability-indicating size exclusion chromatography (SEC) cartridge-based free drug testing method was developed for active loading nanoliposome formulations through a systematic development approach. The SEC spin cartridge columns (7K MWCO, 2 mL) were used for developing the liposome free drug testing procedure. The SEC cartridge column retention capacity for a Mirati drug was determined (445 μg per cartridge). SEC testing conditions were studied to achieve a good separation between liposomes and the free drug, including cartridge conditioning, buffer wash steps for liposomal drug elution, organic media wash steps for free drug elution, and sample size effect. Qualification of this newly developed SEC cartridge method has demonstrated its specificity/selectivity without interference and excellent detection linearity (correlation coefficient (R) > 0.999) over a study concentration range (1.0 to 46.9 μg mL-1), sufficient LOQ (0.52 μg mL-1 equivalent to 1.7% of free drug in a liposome formulation at 2.5 mg mL-1), acceptable accuracy/recovery of 81 to 89% for free drug in spiked samples at 4.5%, 9%, and 18% levels (sample loading by a regular pipet) and 88 to 97% at 10% spike level (sample loading by a positive displacement pipet), good method precision (RSD (n = 6) of 4% NMT) for free drug determination, and 3 days solution stability for both standard and sample solutions (2-8 °C). In comparative sample testing, the SEC free drug test results were in good agreement with the solid phase extraction (SPE) test results for active loading formulations. The new method's stability-indicating ability has been proved through monitoring free drug in a liposome sample stored at -20 °C and -80 °C.

纳米肿瘤药物脂质体制剂中游离药物含量测定方法的建立。
通过系统的开发方法,建立了一种可靠且具有稳定性的粒径排阻色谱(SEC)药筒游离药物检测方法,用于活性负载纳米脂质体制剂。SEC自旋筒柱(7K MWCO, 2 mL)用于开发无脂质体药物测试程序。测定了Mirati药物的SEC药筒柱保留容量(445 μg /药筒)。为了实现脂质体与游离药物的良好分离,研究了SEC测试条件,包括药筒调节、脂质体药物洗脱的缓冲液洗涤步骤、游离药物洗脱的有机介质洗涤步骤和样本量效应。该方法在研究浓度范围(1.0 ~ 46.9 μg mL-1)内具有良好的检测线性(相关系数(R) > 0.999),具有足够的定量限(0.52 μg mL-1,相当于2.5 mg mL-1脂质体制剂中游离药物的1.7%),加标样品中游离药物的可接受准确度/回收率为81 ~ 89%。在18%水平(常规移液器上样)和88%至97%的10%峰值水平(正位移移液器上样)下,游离药物测定的方法精密度良好(RSD (n = 6)为4% NMT),标准溶液和样品溶液(2-8°C)的溶液稳定性均为3天。在对比样品测试中,无SEC药物测试结果与固相萃取(SPE)测试结果吻合良好。在-20℃和-80℃条件下对脂质体样品中的游离药物进行监测,证明了该方法的稳定性指示能力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Analytical Methods
Analytical Methods CHEMISTRY, ANALYTICAL-FOOD SCIENCE & TECHNOLOGY
CiteScore
5.10
自引率
3.20%
发文量
569
审稿时长
1.8 months
期刊介绍: Early applied demonstrations of new analytical methods with clear societal impact
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信