AAPS PharmSciTech

, 20:305 | Cite as

Enhanced the Bioavailability of Sterile 20(S)-Protopanaxadiol Nanocrystalline Suspension Coated by Bovine Serum Albumin for Intramuscular Injection: In Vitro and In Vivo Evaluation

  • Hui Zhang
  • Hongbing Liu
  • Pan Qi
  • Silin Wang
  • Haibin Hu
  • Jingxin Gou
  • Yu Zhang
  • Haibing He
  • Xing Tang
  • Tian YinEmail author
  • Yue YuanEmail author
Research Article


The aim of this study was to prepare a 20(S)-protopanaxadiol nanocrystalline suspension and enhance the bioavailability of 20(S)-protopanaxadiol by intramuscular injection. 20(S)-Protopanaxadiol nanocrystalline suspension was prepared using an anti-solvent combined with ultrasonic approach, in which meglumine and bovine serum albumin were screened as the optimized stabilizer and the coating agent during spray drying process, respectively. The optimal nanocrystallines were nearly spherical with a uniform particle size distribution, the mean particle size, polydispersity index, and drug loading of which were 151.20 ± 2.54 nm, 0.11 ± 0.01, and 47.15% (w/w), respectively. Sterile 20(S)-protopanaxadiol nanocrystalline suspension was obtained by passing through a 0.22-μm membrane, and the average filtration efficiency (FE%) was 99.96%. The cumulative release percentage of 20(S)-protopanaxadiol nanocrystalline suspension was 92.36% 20(S)-protopanaxadiol within 60 min in vitro, which was relatively rapid compared with that of the physical mixture for 12.51% and the 20(S)-protopanaxadiol bulk powder for 9.71% during the same time interval. The sterile 20(S)-protopanaxadiol nanocrystalline suspension caused minimal irritation responses by histological examination, indicating a good biocompatibility between the 20(S)-protopanaxadiol nanocrystalline suspension and muscle tissues. In pharmacokinetic study, the absolute bioavailability of 20(S)-protopanaxadiol nanocrystalline suspension for intramuscular injection and for oral gavage was 5.99 and 0.03, respectively. In summary, the 20(S)-protopanaxadiol nanocrystalline via intramuscular injection is an efficient drug delivery system to improve its bioavailability.


nanocrystallines intramuscular injection oral gavage biocompatibility bioavailability 



We thank Amanda Pearce’s correction for the manuscript.

Compliance with Ethical Standards

All animal experiments were in accordance with the requirements of animal ethical.

Conflict of Interest

The authors declare that they have no conflicts of interest.


  1. 1.
    D-q L, Cheng Z-q, Q-j F, H-j L, S-f Y, Teng B. Polycaprolactone nanofibres loaded with 20 (S)-protopanaxadiol for in vitro and in vivo anti-tumour activity study. Royal Society Open Science. 2018;5(5):180137.CrossRefGoogle Scholar
  2. 2.
    Jin HR, Du CH, Wang CZ, Yuan CS, Du W. Ginseng metabolite protopanaxadiol interferes with lipid metabolism and induces endoplasmic reticulum stress and p53 activation to promote cancer cell death. Phytother Res. 2019;33(3):610–7.PubMedGoogle Scholar
  3. 3.
    Zhang Q, Pu Y, Wang B, Wang Y, Dong T, Guo T, et al. Characterization, molecular docking, and in vitro dissolution studies of solid dispersions of 20 (S)-protopanaxadiol. Molecules. 2017;22(2):274.CrossRefGoogle Scholar
  4. 4.
    Chen C, Wang L, Cao F, Miao X, Chen T, Chang Q, et al. Formulation of 20(S)-protopanaxadiol nanocrystals to improve oral bioavailability and brain delivery. Int J Pharm. 2016;497(1–2):239–47.CrossRefGoogle Scholar
  5. 5.
    Jin X, Zhang ZH, Sun E, Liu QY, Jia XB. Study on pharmacokinetics of 20(S)-protopanaxadiol lipid cubic nanoparticles. China Journal of Chinese Materia Medica. 2013;38(2):263–8.PubMedGoogle Scholar
  6. 6.
    Xin J, Zhang ZH, Li SL, Sun E, Tan XB, Jie S, et al. A nanostructured liquid crystalline formulation of 20(S)-protopanaxadiol with improved oral absorption. Fitoterapia. 2013;84(1):64–71.Google Scholar
  7. 7.
    Xia HJ, Zhang Z-h, Jin H, Chen J X-b. A novel drug-phospholipid complex enriched with micelles: preparation and evaluation in vitro and in vivo. Int J Nanomedicine. 2013;8(10):545–54.CrossRefGoogle Scholar
  8. 8.
    Han M, Chen J, Chen S, Wang X. Development of a UPLC-ESI-MS/MS assay for 20(S)-protopanaxadiol and pharmacokinetic application of its two formulations in rats. Analytical Sciences the International Journal of the Japan Society for Analytical Chemistry. 2010;26(7):749–53.CrossRefGoogle Scholar
  9. 9.
    Han M, Wang Y. Determination of 20(S)-protopanaxadiol in rat plasma by LC-MS/MS and its application to the pharmacokinetic study: a comparative study of its solution and two oral formulations. J Anal Chem. 2013;68(8):730–5.CrossRefGoogle Scholar
  10. 10.
    Han M, Jing C, Wang Y, Chen S, Wang X. Determination of 20(S)-protopanaxadiol in rat plasma by LC-MS/MS and its application to the pharmacokinetic study: a comparative study of its solution and two oral formulations. J Anal Chem. 2013;68(8):730–5.CrossRefGoogle Scholar
  11. 11.
    Waard HD, Frijlink HW, Hinrichs WLJ. Bottom-up preparation techniques for nanocrystals of lipophilic drugs. Pharm Res. 2011;28(5):1220–3.CrossRefGoogle Scholar
  12. 12.
    Hu X, Chen X, Zhang L, Lin X, Zhang Y, Tang X, et al. A combined bottom–up/top–down approach to prepare a sterile injectable nanosuspension. Int J Pharm. 2014;472(1–2):130–9.CrossRefGoogle Scholar
  13. 13.
    Dalvi SV, Dave RN. Controlling particle size of a poorly water-soluble drug using ultrasound and stabilizers in antisolvent precipitation. Ind Eng Chem Res. 2009;48(16):7581–93.CrossRefGoogle Scholar
  14. 14.
    Sommerfeld P, Schroeder U, Sabel BA. Sterilization of unloaded polybutylcyanoacrylate nanoparticles. International Journal of Pharmaceutics. 1998;164(s 1–2):113–8.CrossRefGoogle Scholar
  15. 15.
    El-Salamouni NS, Farid RM, El-Kamel AH, El-Gamal SS. Effect of sterilization on the physical stability of brimonidine-loaded solid lipid nanoparticles and nanostructured lipid carriers. Int J Pharm. 2015;496(2):976–83.CrossRefGoogle Scholar
  16. 16.
    Johnson NJ, Korinek A, Dong C, van Veggel FC. Self-focusing by Ostwald ripening: a strategy for layer-by-layer epitaxial growth on upconverting nanocrystals. J Am Chem Soc. 2012;134(27):11068–71.CrossRefGoogle Scholar
  17. 17.
    Adler M, Unger M, Lee G. Surface composition of spray-dried particles of bovine serum albumin/trehalose/surfactant. Pharm Res. 2000;17(7):863–70.CrossRefGoogle Scholar
  18. 18.
    Li FQ, Hu JH, Lu B, Yao H, Zhang WGF. Ciprofloxacin-loaded bovine serum albumin microspheres: preparation and drug-release in vitro. J Microencapsul. 2001;18(6):825–9.CrossRefGoogle Scholar
  19. 19.
    Cai Q, He L, Wang S, Chu W, Zhou L, Pan W, et al. Process control and in vitro/in vivo evaluation of aripiprazole sustained-release microcrystals for intramuscular injection. Eur J Pharm Sci. 125:193–204.CrossRefGoogle Scholar
  20. 20.
    Liu Z, Yang L. Antisolvent precipitation for the preparation of high polymeric procyanidin nanoparticles under ultrasonication and evaluation of their antioxidant activity in vitro. Ultrason Sonochem. 2018;43:208–18.CrossRefGoogle Scholar
  21. 21.
    Igawa T, Kameoka D. Stabilizer for protein preparation comprising meglumine and use thereof. Google Patents; 2015.Google Scholar
  22. 22.
    Aminzadeh S, Lauberts M, Dobele G, Ponomarenko J, Mattsson T, Lindström ME, et al. Membrane filtration of kraft lignin: structural charactristics and antioxidant activity of the low-molecular-weight fraction. Ind Crop Prod. 2018;112:200–9.CrossRefGoogle Scholar
  23. 23.
    Vieira GN, Olazar M, Freire JT, Freire FB. Real-time monitoring of milk powder moisture content during drying in a spouted bed dryer using a hybrid neural soft sensor. Dry Technol 2018;37(9):1–7.CrossRefGoogle Scholar
  24. 24.
    Razak N, Hamid N, Shaari A, editors. Effect of storage temperature on moisture content of encapsulated Orthosiphon stamineus spray-dried powder. AIP Conference Proceedings; 2018: AIP Publishing.Google Scholar
  25. 25.
    Fischer W, Wieland G, Krenn KD. Karl Fischer reagent and method for the determination of water using this reagent 1985.Google Scholar
  26. 26.
    Hoo CM, Starostin N, West P, Mecartney ML. A comparison of atomic force microscopy (AFM) and dynamic light scattering (DLS) methods to characterize nanoparticle size distributions. J Nanopart Res. 2008;10(1):89–96.CrossRefGoogle Scholar
  27. 27.
    Leng D, Chen H, Li G, Guo M, Zhu Z, Xu L, et al. Development and comparison of intramuscularly long-acting paliperidone palmitate nanosuspensions with different particle size. Int J Pharm. 2014;472(1–2):380–5.CrossRefGoogle Scholar
  28. 28.
    Ali HS, York PN. Preparation of hydrocortisone nanosuspension through a bottom-up nanoprecipitation technique using microfluidic reactors. Int J Pharm. 2009;375(1):107–13.CrossRefGoogle Scholar
  29. 29.
    Botha N, Wessels G, Botha N, van Eden B, editors. Image processing towards the automated identification of nanoparticles in SEM images. 2019 Southern African Universities Power Engineering Conference/Robotics and Mechatronics/Pattern Recognition Association of South Africa (SAUPEC/RobMech/PRASA); 2019: IEEE.Google Scholar
  30. 30.
    Majoinen J, Kontturi E, Ikkala O, Gray DG. SEM imaging of chiral nematic films cast from cellulose nanocrystal suspensions. Cellulose. 2012;19(5):1599–605.CrossRefGoogle Scholar
  31. 31.
    Panthani MG, Hessel CM, Reid D, Casillas G, Korgel BA. Graphene-supported high-resolution TEM and STEM imaging of silicon nanocrystals and their capping ligands. J Phys Chem C. 2012;116(116):22463–8.CrossRefGoogle Scholar
  32. 32.
    Li L, Li W, Sun J, Zhang H, Gao J, Guo F, et al. Preparation and evaluation of progesterone nanocrystals to decrease muscle irritation and improve bioavailability. AAPS PharmSciTech. 2018;19(3):1254–63.CrossRefGoogle Scholar
  33. 33.
    Yu X. A nanoparticulate drug-delivery system for 20(S)-protopanaxadiol: formulation, characterization, increased oral bioavailability and anti-tumor efficacy. Drug Delivery. 2016;23(7):1–9.Google Scholar
  34. 34.
    Ben-Eltriki M, Hassona M, Meckling G, Adomat H, Deb S, Guns EST. Pharmacokinetic interaction of calcitriol with 20 (S)-protopanaxadiol in mice: determined by LC/MS analysis. Eur J Pharm Sci. 2019;130:173–80.CrossRefGoogle Scholar
  35. 35.
    Luo Y, Xu L, Xu M, Tao X, Ai R, Tang X. Improvement of dissolution and bioavailability of Ginsenosides by hot melt extrusion and cogrinding. Drug Development & Industrial Pharmacy. 2013;39(1):109–16.CrossRefGoogle Scholar
  36. 36.
    Darniadi S, Ho P, Murray BS. Comparison of blueberry powder produced via foam-mat freeze-drying versus spray-drying: evaluation of foam and powder properties. J Sci Food Agric. 2018;98(5):2002–10.CrossRefGoogle Scholar
  37. 37.
    Hu L, Yang C, Kong D, Hu Q, Na G, Zhai F. Development of a long-acting intramuscularly injectable formulation with nanosuspension of andrographolide. Journal of Drug Delivery Science & Technology. 2016;35:327–32.CrossRefGoogle Scholar
  38. 38.
    Jiazhuo W, Mackenzie JD, Rageshree R, Chen DZ. Identifying neutrophils in H&E staining histology tissue images. 2014;8673:73–80.Google Scholar
  39. 39.
    Sosnik A, Seremeta KP. Advantages and challenges of the spray-drying technology for the production of pure drug particles and drug-loaded polymeric carriers. Advances in Colloid & Interface Science. 2015;223:40–54.CrossRefGoogle Scholar
  40. 40.
    Fejzo J, Lepre CA, Peng JW, Bemis GW, Ajay, Murcko MA, et al. The SHAPES strategy: an NMR-based approach for lead generation in drug discovery. Chem Biol. 1999;6(10):755–69.CrossRefGoogle Scholar
  41. 41.
    Céline H, Sophie L, Muller RN, Luce VE. HR-MAS NMR spectroscopy: an innovative tool for the characterization of iron oxide nanoparticles tracers for molecular imaging. Anal Chem. 2015;87(3):1701–10.CrossRefGoogle Scholar
  42. 42.
    Kojima T, Karashima M, Yamamoto K, Ikeda Y. A combination of NMR methods to reveal the interfacial structure of a pharmaceutical nanocrystal and nanococrystal in the suspended state. Mol Pharm. 2018;15(9):3901–8.CrossRefGoogle Scholar
  43. 43.
    Darville N, Heerden MV, Mariën D, Meulder MD, Rossenu S, An V, et al. The effect of macrophage and angiogenesis inhibition on the drug release and absorption from an intramuscular sustained-release paliperidone palmitate suspension. J Control Release. 2016;230:95–108.CrossRefGoogle Scholar
  44. 44.
    Fernández-Varón E, Bovaira MJ, Espuny A, Escudero E, Vancraeynest D, Cárceles CM. Pharmacokinetic-pharmacodynamic integration of moxifloxacin in rabbits after intravenous, intramuscular and oral administration. J Veterinary Pharmacol Ther. 2010;28(4):343–8.CrossRefGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2019

Authors and Affiliations

  • Hui Zhang
    • 1
  • Hongbing Liu
    • 1
  • Pan Qi
    • 1
  • Silin Wang
    • 2
  • Haibin Hu
    • 3
  • Jingxin Gou
    • 1
  • Yu Zhang
    • 1
  • Haibing He
    • 1
  • Xing Tang
    • 1
  • Tian Yin
    • 4
    • 5
    Email author
  • Yue Yuan
    • 1
    Email author
  1. 1.School of PharmacyShenyang Pharmaceutical UniversityShenyangPeople’s Republic of China
  2. 2.School of Traditional Chinese Materia MedicaShenyang Pharmaceutical UniversityShenyangPeople’s Republic of China
  3. 3.Wuya College of InnovationShenyang Pharmaceutical UniversityShenyangPeople’s Republic of China
  4. 4.School of Functional Food and WineShenyang Pharmaceutical UniversityShenyangPeople’s Republic of China
  5. 5.Jiangsu Kanion Pharmaceutical Co., Ltd.LianyungangPeople’s Republic of China

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