Skip to main content

High-Performance Liquid Chromatography Coupled with Tandem Mass Spectrometry Method for the Identification and Quantification of Lipids in Liposomes

  • Protocol
  • First Online:
Liposomes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2622))

Abstract

Liposomes are spherical, closed vesicles consisting of at least one lipid bilayer with a water chamber and are widely used to encapsulate bioactive molecules. Lipid membranes, composed of different types of lipids or lipophilic components, determine whether liposomes can achieve the desired purpose and determine the overall quality of liposomes. Thus, the quantification of lipid components and encapsulated molecules is essential to characterize and control the quality of liposomes. Moreover, multicomponent simultaneous determination is the preferred method for lipid component analysis in liposomes. Therefore, the present work describes an analytical methodology for the simultaneous determination of commonly used lipids in liposome formulations, using high-performance liquid chromatography coupled with a tandem mass spectrometry (MS) detector (HPLC-MS/MS). HPLC-MS/MS consists of a rapid and highly efficient chromatographic separation of the liposomal components with a C18 column and the subsequent detection of the ingredients through an MS detector, along with an accurate mass fragmentation pattern. The analytical process mainly includes lipid extraction, solution preparation, the optimization of chromatographic conditions, and method validation. We hope this analytical methodology is valuable and efficient and can be applied to the analysis of multiple types of lipids in liposomes, such as raw material quality analysis, formulation study, overall quality control, etc.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 249.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Mehdi Hasan MH, Mondal JC, Al Hasan M, Talukder S, Rashid HA (2017) Liposomes: an advance tools for novel drug delivery system. Pharma Innov J 6:304–311

    Google Scholar 

  2. Petros RA, DeSimone JM (2010) Strategies in the design of nanoparticles for therapeutic applications. Nat Rev Drug Discov 9:615–627

    Article  CAS  Google Scholar 

  3. Akbarzadeh A, Rezaei-Sadabady R, Davaran S, Joo SW, Zarghami N, Hanifehpour Y et al (2013) Liposome: classification, preparation, and applications. Nanoscale Res Lett 8(1):102

    Article  Google Scholar 

  4. Soema PC, Willems GJ, Jiskoot W, Amorij JP, Kersten GF (2015) Predicting the influence of liposomal lipid composition on liposome size, zeta potential and liposome-induced dendritic cell maturation using a design of experiments approach. Eur J Pharm Biopharm 94:427–435

    Article  CAS  Google Scholar 

  5. Anderson M, Omri A (2004) The effect of different lipid components on the in vitro stability and release kinetics of liposome formulations. Drug Deliv 11:33–39

    Article  CAS  Google Scholar 

  6. He H, Lu Y, Qi J, Zhu Q, Chen Z, Wu W (2019) Adapting liposomes for oral drug delivery. Acta Pharm Sin B 9:36–48

    Article  Google Scholar 

  7. Yandrapati RK (2012) Effect of lipid composition on the physical properties of liposomes: a light scattering study. Masters Theses 6864

    Google Scholar 

  8. Poelma DL, Ju MR, Bakker SC, Zimmermann LJ, Lachmann BF, van Iwaarden JF (2004) A common pathway for the uptake of surfactant lipids by alveolar cells. Am J Respir Cell Mol Biol 30:751–758

    Article  CAS  Google Scholar 

  9. Ciani L, Casini A, Gabbiani C, Ristori S, Messori L, Martini G (2007) DOTAP/DOPE and DC-Chol/DOPE lipoplexes for gene delivery studied by circular dichroism and other biophysical techniques. Biophys Chem 127:213–220

    Article  CAS  Google Scholar 

  10. Briuglia ML, Rotella C, McFarlane A, Lamprou DA (2015) Influence of cholesterol on liposome stability and on in vitro drug release. Drug Deliv Transl Res 5:231–242

    Article  CAS  Google Scholar 

  11. Kaddah S, Khreich N, Kaddah F, Charcosset C, Greige-Gerges H (2018) Cholesterol modulates the liposome membrane fluidity and permeability for a hydrophilic molecule. Food Chem Toxicol 113:40–48

    Article  CAS  Google Scholar 

  12. Haeri A, Alinaghian B, Daeihamed M, Dadashzadeh S (2014) Preparation and characterization of stable nanoliposomal formulation of fluoxetine as a potential adjuvant therapy for drug-resistant tumors. Iran J Pharm Res 13:3–14

    CAS  Google Scholar 

  13. Caracciolo G, Pozzi D, Capriotti AL, Cavaliere C, Piovesana S, Amenitsch H et al (2015) Lipid composition: a “key factor” for the rational manipulation of the liposome–protein corona by liposome design. RSC Adv 5:5967–5975

    Article  CAS  Google Scholar 

  14. Liposome drug products – chemistry, manufacturing, and controls; Human pharmacokinetics and bioavailability; and labeling documentation – guidance for industry. 2015

    Google Scholar 

  15. Sophia Hatziantoniou CD (2010) Method of simultaneous analysis of liposome components using HPTLC/FID. Methods Mol Biol 606:363–368

    Article  Google Scholar 

  16. Sophia Hatziantoniou CD (2017) Method of simultaneous analysis of liposome components using HPTLC/FID. Methods Mol Biol 1522:49–54

    Article  Google Scholar 

  17. Hatziantoniou S, Demetzos C (2006) Qualitative and quantitative one-step analysis of lipids and encapsulated bioactive molecules in liposome preparations by HPTLC/FID (IATROSCAN). J Liposome Res 16:321–330

    Article  CAS  Google Scholar 

  18. Suchocka Z, Gronostajska D, Suchocki P, Pachecka J (2003) New HPLC method for separation of blood plasma phospholipids. J Pharm Biomed Anal 32:859–865

    Article  CAS  Google Scholar 

  19. Singh R, Ajagbe M, Bhamidipati S, Ahmad Z, Ahmad I (2005) A rapid isocratic high-performance liquid chromatography method for determination of cholesterol and 1,2-dioleoyl-sn-glycero-3-phosphocholine in liposome-based drug formulations. J Chromatogr A 1073:347–353

    Article  CAS  Google Scholar 

  20. Hvattum E, Uran S, Sandbaek AG, Karlsson AA, Skotland T (2006) Quantification of phosphatidylserine, phosphatidic acid and free fatty acids in an ultrasound contrast agent by normal-phase high-performance liquid chromatography with evaporative light scattering detection. J Pharm Biomed Anal 42:506–512

    Article  CAS  Google Scholar 

  21. Alsaadi MM, Christine Carter K, Mullen AB (2013) High performance liquid chromatography with evaporative light scattering detection for the characterisation of a vesicular delivery system during stability studies. J Chromatogr A 1320:80–85

    Article  CAS  Google Scholar 

  22. Kothalawala N, Mudalige TK, Sisco P, Linder SW (2018) Novel analytical methods to assess the chemical and physical properties of liposomes. J Chromatogr B 1091:14–20

    Article  CAS  Google Scholar 

  23. Lesnefsky EJ, Stoll MS, Minkler PE, Hoppel CL (2000) Separation and quantitation of phospholipids and lysophospholipids by high-performance liquid chromatography. Anal Biochem 285:246–254

    Article  CAS  Google Scholar 

  24. Siriwardane DA, Wang C, Jiang W, Mudalige T (2020) Quantification of phospholipid degradation products in liposomal pharmaceutical formulations by ultra performance liquid chromatography-mass spectrometry (UPLC-MS). Int J Pharm 578:119077

    Article  CAS  Google Scholar 

  25. Wang C, Siriwardane DA, Jiang W, Mudalige T (2019) Quantitative analysis of cholesterol oxidation products and desmosterol in parenteral liposomal pharmaceutical formulations. Int J Pharm 569:118576

    Article  CAS  Google Scholar 

  26. Jeschek D, Lhota G, Wallner J, Vorauer-Uhl K (2016) A versatile, quantitative analytical method for pharmaceutical relevant lipids in drug delivery systems. J Pharm Biomed Anal 119:37–44

    Article  CAS  Google Scholar 

  27. Meyer O, Roch O, Elmlinger D, Kolbe HVJ (2000) Direct lipid quantitation of cationic liposomes by reversed-phase HPLC in lipoplex preparation process. Eur J Pharm Biopharm 50:353–356

    Article  CAS  Google Scholar 

  28. Simonzadeh N (2009) An isocratic HPLC method for the simultaneous determination of cholesterol, Cardiolipin, and DOPC in lyophilized lipids and liposomal formulations. J Chromatogr Sci 47:304–308

    Google Scholar 

  29. Oswald M, Platscher M, Geissler S, Goepferich A (2016) HPLC analysis as a tool for assessing targeted liposome composition. Int J Pharm 497:293–300

    Article  CAS  Google Scholar 

  30. Zhou Q, Liu L, Zhang D, Fan X (2012) Analysis of gemcitabine liposome injection by HPLC with evaporative light scattering detection. J Liposome Res 22:263–269

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yujie Shi .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Shi, Y., Li, X. (2023). High-Performance Liquid Chromatography Coupled with Tandem Mass Spectrometry Method for the Identification and Quantification of Lipids in Liposomes. In: D'Souza, G.G., Zhang, H. (eds) Liposomes. Methods in Molecular Biology, vol 2622. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2954-3_20

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-2954-3_20

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2953-6

  • Online ISBN: 978-1-0716-2954-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics