• ELIZA MIRANDA Department of Dermatology and Venereology, Faculty of Medicine, Universitas Indonesia, Depok, Indonesia
  • KUSMARINAH BRAMONO Department of Dermatology and Venereology, Faculty of Medicine, Universitas Indonesia, Depok, Indonesia
  • LUDDWI ACHMAD RIZKY Faculty of Medicine, Universitas Indonesia, Depok, Indonesia
  • HAYUN Faculty of Pharmacy, Universitas Indonesia, Depok, Indonesia



LL-37, Cream, High-performance liquid chromatograph, Stability


Objective: The present study aimed to prepare LL-37 in a cream formulation (O/W emulsion) and evaluate its stability by determining the physical changes in the cream and concentration of LL-37 using validated high-performance liquid chromatography.

Methods: The method was conducted at room temperature using a C18 column (5 µm × 250 mm × 4.6 mm) as a stationary phase, a mixture of 0.1% trifluoroacetic acid (TFA)/water (A) and 0.1% TFA/acetonitrile (B) (85:15) as the mobile phase, a flow rate of 1.0 mL/min, and photodiode array set at 228 nm as the detector. The method was validated in compliance with the Association of Official Analytical Chemists and International Conference on Harmonization guidelines. It demonstrated excellent linearity, accuracy, precision, specificity, the limit of detection, and limit of quantitation.

Results: The chromatographic analysis indicated minimal degradation of LL-37 during the 12-week, with a predicted expiry time of 99 and 75 mo stored at 4 °C and 28 °C, respectively.

Conclusion: LL-37 cream establishes good physical characteristics and stabilizes the active ingredient, especially at 4 °C and 28 °C storage. Therefore, the emulsion delivery system of LL-37 cream is harmless and stable as a novel alternative vehicle of LL-37.


Download data is not yet available.


Cho NH, Shaw JE, Karuranga S, Huang Y, da Rocha Fernandes JD, Ohlrogge AW, Malanda B. IDF Diabetes Atlas: global estimates of diabetes prevalence for 2017 and projections for 2045. Diabetes Res Clin Pract. 2018;138:271-81. doi: 10.1016/j.diabres.2018.02.023, PMID 29496507.

Alavi A, Sibbald RG, Mayer D, Goodman L, Botros M, Armstrong DG, Woo K, Boeni T, Ayello EA, Kirsner RS. Diabetic foot ulcers: Part II. Management. J Am Acad Dermatol. 2014;70(1):21.e1-24; quiz 45. doi: 10.1016/j.jaad.2013.07.048, PMID 24355276.

Mohd KS, Hassan MA, Azemin W, Dharmaraj S. A review of potential anticancers from antimicrobial peptides. Int J Pharm Pharm Sci. 2014;7:19-26.

Frohm Nilsson MF, Sandstedt B, Sørensen O, Weber G, Borregaard N, Stahle-Backdahl M. The human cationic antimicrobial protein (hCAP18), a peptide antibiotic, is widely expressed in human squamous epithelia and colocalizes with interleukin-6. Infect Immun. 1999;67(5):2561-6. doi: 10.1128/IAI.67.5.2561-2566.1999, PMID 10225921.

Dean SN, Bishop BM, van Hoek ML. Natural and synthetic cathelicidin peptides with anti-microbial and anti-biofilm activity against Staphylococcus aureus. BMC Microbiol. 2011;11:114. doi: 10.1186/1471-2180-11-114, PMID 21605457.

Heilborn JD, Nilsson MF, Kratz G, Weber G, Sorensen O, Borregaard N, Stahle-Backdahl M. The cathelicidin anti-microbial peptide LL-37 is involved in re-epithelialization of human skin wounds and is lacking in chronic ulcer epithelium. J Invest Dermatol. 2003;120(3):379-89. doi: 10.1046/j.1523-1747.2003.12069.x, PMID 12603850.

Ramos R, Silva JP, Rodrigues AC, Costa R, Guardão L, Schmitt F, Soares R, Vilanova M, Domingues L, Gama M. Wound healing activity of the human antimicrobial peptide LL37. Peptides. 2011;32(7):1469-76. doi: 10.1016/j.peptides.2011.06.005, PMID 21693141.

Steinstraesser L, Ring A, Bals R, Steinau HU, Langer S. The human host defense peptide LL37/hCAP accelerates angiogenesis in PEGT/PBT biopolymers. Ann Plast Surg. 2006;56(1):93-8. doi: 10.1097/, PMID 16374104.

Grönberg A, Mahlapuu M, Ståhle M, Whately-Smith C, Rollman O. Treatment with LL-37 is safe and effective in enhancing healing of hard-to-heal venous leg ulcers: a randomized, placebo-controlled clinical trial. Wound Repair Regen. 2014;22(5):613-21. doi: 10.1111/wrr.12211, PMID 25041740.

Grönberg A, Zettergren L, Agren MS. Stability of the cathelicidin peptide LL-37 in a non-healing wound environment. Acta Derm Venereol. 2011;91(5):511-5. doi: 10.2340/00015555-1102, PMID 21547341.

Krahulec J, Hyrsova M, Pepeliaev S, Jilkova J, Cerny Z, Machalkova J. High-level expression and purification of antimicrobial human cathelicidin LL-37 in Escherichia coli. Appl Microbiol Biotechnol. 2010;88(1):167-75. doi: 10.1007/s00253-010-2736-7, PMID 20625720.

Association of Official Agricultural Chemists. Official methods of analysis. 16th ed. Rockville: AOAC; 2002.

International Conference of Harmonisation. Validation of analytical procedures: text and methodology. Vol. Q2. Switzerland: ICH; 2005. p. R1.

Grimm W. Stability testing of clinical trial materials. In: Rhodes CT, Carstensen JT, editors. Drugs and the pharmaceutical sciences. 3rd ed. rev and expanded ed. New York: Marcel Dekker; 2000.

Hong DD, Shah M. Development and validation of HPLC stability-indicating assays. In: Rhodes CT, Carstensen JT, editors. Drugs and the pharmaceutical sciences. 3rd ed, rev and expanded ed. New York: Marcel Dekker; 2000.

Chen Y, Yang S, Ho EA. Development of an analytical method for the rapid quantitation of peptides used in microbicide formulations. Chromatographia. 2014;77(23-24):1713-20. doi: 10.1007/s10337-014-2777-7, PMID 25477555.

Smaoui S, Ben Hlima H, Ben Chobba I, Kadri A. Development and stability studies of sunscreen cream formulations containing three photo-protective filters. Arab J Chem. 2017;10:S1216-22. doi: 10.1016/j.arabjc.2013.02.020.

Anwar S, Jan SU, Gul R. Formulation and evaluation of celecoxib cream and its released study. Int J Curr Pharm Sci. 2020;12:15-9. doi: 10.22159/ijcpr.2020v12i5.39757.

Sirisa Ard P, Peerakam N, Huy NQ, On TV, Long PT, Intharuksa A. Development of anti-wrinkle cream from Pueraria candollei var. mirifica (airy shaw and suvat.) niyomdham, “Kwao krua kao” for menopausal women. Int J Pharm Pharm Sci. 2018;10(7):16-21. doi: 10.22159/ijpps.2018v10i7.24665.



How to Cite

MIRANDA, E., BRAMONO, K., RIZKY, L. A., & HAYUN. (2021). PREPARATION AND STABILITY EVALUATION OF LL-37 CREAM. International Journal of Applied Pharmaceutics, 13(6), 139–143.



Original Article(s)