, Volume 79, Issue 13–14, pp 851–860 | Cite as

Simultaneous Detection of Glabridin, (−)-α-Bisabolol, and Ascorbyl Tetraisopalmitate in Whitening Cosmetic Creams Using HPLC-PAD

  • Jong-Sup JeonEmail author
  • Han-Taek Kim
  • Myeong-Gil Kim
  • Moon-Seog Oh
  • Se-Ra Hong
  • Mi-Hye Yoon
  • Ho-Chul Shin
  • Jae-Han Shim
  • Nehal Aly Afifi
  • Ahmet Hacımüftüoğlu
  • A. M. Abd El-AtyEmail author


A simultaneous analytical method was developed and validated to quantify three lipophilic compounds; namely glabridin (an isoflavonoid isolated from crude licorice), (−)-α-bisabolol (a sesquiterpene alcohol obtained from plant extracts), and ascorbyl tetraisopalmitate (a fat-soluble molecule derived from vitamin C) in functional cosmetic cream using high-performance liquid chromatography (HPLC) coupled with photodiode array detection (PAD). Cosmetic cream samples were extracted with a mixture of acetonitrile and isopropyl alcohol (45:55, v/v) and the target compounds were separated on a C18 column with a gradient mobile phase consisting of deionized water, acetonitrile, and isopropyl alcohol. The detector wavelengths were 228, 202, and 221 nm, for glabridin, (−)-α-bisabolol, and ascorbyl tetraisopalmitate, respectively. The calibration curves showed good linearity with determination coefficients (R 2) ≥ 0.999. The mean recoveries were ranged between 89.8 and 103.9 % with relative standard deviations (RSDs) <5 %. The intra- and inter-day precision was <2 %. The limits of detection (LODs) were 0.03, 0.4, and 4.02 μg mL−1 for glabridin, (−)-α-bisabolol, and ascorbyl tetraisopalmitate, respectively. The method was successfully applied for monitoring 11 market samples, in which glabridin was quantified in the range of 17.5–25 mg 100 g−1, (−)-α-bisabolol in the range of 25.1–677 mg 100 g−1, and 140.6–291.5 mg 100 g−1 for ascorbyl tetraisopalmitate. The proposed analytical method is simple, sensitive, and versatile and can be used for the quantification of lipophilic compounds in cosmetics in a single chromatographic run.


High-performance liquid chromatography Glabridin (−)-α-Bisabolol Ascorbyl tetraisopalmitate Functional cosmetic 



The authors gratefully acknowledge the financial support from the Gyeonggi Province Institute of Health and Environment (Suwon city, Republic of Korea).

Compliance with standards

Conflict of interest

The authors have declared no conflict of interest.


  1. 1.
    Korea Ministry of drug and food safety (2013) Cosmetic act, Seoul, Republic of KoreaGoogle Scholar
  2. 2.
    Gao XH, Zhang L, Wei H, Chen HD (2008) Efficacy and safety of innovative cosmeceuticals. Clin Dermatol 26:367–374CrossRefGoogle Scholar
  3. 3.
    Amer M, Maged M (2009) Cosmeceuticals versus pharmaceuticals. Clin Dermatol 27:428–430CrossRefGoogle Scholar
  4. 4.
    Jeon JS, Kim HT, Kim MG, Oh MS, Hong SR, Yoon MH, Cho SM, Shin HC, Shim JH, Ramadan A, Abd El-Aty AM (2016) Simultaneous determination of water-soluble whitening ingredients and adenosine in different cosmetic formulations by high-performance liquid chromatography coupled with photodiode array detection. Int J Cosmet Sci 38:286–293CrossRefGoogle Scholar
  5. 5.
    Chang ML, Chang CM (2003) Simultaneous HPLC determination of hydrophilic whitening agents in cosmetic products. J Pharm Biomed Anal 33:617–626CrossRefGoogle Scholar
  6. 6.
    Jain P, Sonti S, Garruto J, Mehta R, Banga AK (2012) Formulation optimization, skin irritation, and efficacy characterization of a novel skin-lightening agent. J Cosmet Dermatol 11:101–110CrossRefGoogle Scholar
  7. 7.
    Rendon M, Berneburg M, Arellano I, Picardo M (2006) Treatment of melasma. J Am Acad Dermatol 54:272–281CrossRefGoogle Scholar
  8. 8.
    Huang SC, Lin CC, Huang MC, Wen KC (2004) Simultaneous determination of magnesium ascorbyl phosphate, ascorbyl glucoside, kojic acid, arbutin and hydroquinone in skin whitening cosmetics by HPLC. J Food Drug Anal 12:13–18Google Scholar
  9. 9.
    Jin W, Wang WY, Zhang YL, Yang YJ, Chu QC, Ye JN (2013) Determination of phenolic whitening agents in cosmetics by micellar electrokinetic capillary chromatography with amperometric detection. Chin Chem Lett 24:636–638CrossRefGoogle Scholar
  10. 10.
    Jeon JS, Kim BH, Lee SH, Kwon HJ, Bae HJ, Kim SK, Park JA, Shim JH, Abd El-Aty AM, Shin HC (2015) Simultaneous determination of arbutin and its decomposed product hydroquinone in whitening creams using high-performance liquid chromatography with photodiode array detection: effect of temperature and pH on decomposition. Int J Cosmet Sci 37:567–573CrossRefGoogle Scholar
  11. 11.
    Korea Ministry of drug and food safety (2013) Korean Functional Cosmetics Codex, Seoul, Republic of KoreaGoogle Scholar
  12. 12.
    Jeon JS, Lee MJ, Yoon MH, Park JA, Yi H, Cho HJ, Shin HC (2014) Determination of arbutin, niacinamide, and adenosine in functional cosmetic products by high-performance liquid chromatography. Anal Lett 47:1650–1660CrossRefGoogle Scholar
  13. 13.
    Simmler C, Pauli GF, Chen S-N (2013) Phytochemistry and biological properties of glabridin. Fitoterapia 90:160–184CrossRefGoogle Scholar
  14. 14.
    Yokota T, Nishio H, Kubota Y, Mizoguchi M (1998) The inhibitory effect of glabridin from licorice extracts on melanogenesis and inflammation. Pigment Cell Res 11:355–361CrossRefGoogle Scholar
  15. 15.
    Kamal YT, Singh M, Tamboli ET, Parveen R, Zaidi SM, Ahmad S (2012) Rapid RP-HPLC method for the quantification of glabridin in crude drug and in polyherbal formulation. J Chromatogr Sci 50:779–784Google Scholar
  16. 16.
    Rosenblat M, Belinky P, Vaya J, Levy R, Hayek T, Coleman R, Merchav S, Aviram M (1999) Macrophage enrichment with the isoflavan glabridin inhibits NADPH oxidase-induced cell-mediated oxidation of low density lipoprotein a possible role for protein kinase c. J Biol Chem 274:13790–13799CrossRefGoogle Scholar
  17. 17.
    Waleczek KJ, Marques HMC, Hempel B, Schmidt PC (2003) Phase solubility studies of pure (−)-α-bisabolol and camomile essential oil with β-cyclodextrin. Eur J Pharm Biopharm 55:247–251CrossRefGoogle Scholar
  18. 18.
    Murugan R, Mallavarapu GR (2013) α-Bisabolol, the main constituent of the essential oil of Pogostemon speciosus. Ind Crops Prod 49:237–239CrossRefGoogle Scholar
  19. 19.
    Bhatia SP, McGinty D, Letizia CS, Api AM (2008) Fragrance material review on α-bisabolol. Food Chem Toxicol 46:72–76CrossRefGoogle Scholar
  20. 20.
    Vila R, Santana AI, Pérez-Rosés R, Valderrama A, Castelli MV, Mendonca S, Zacchino S, Gupta MP, Cañigueral S (2010) Composition and biological activity of the essential oil from leaves of Plinia cerrocampanensis, a new source of α-bisabolol. Bioresour Technol 101:2510–2514CrossRefGoogle Scholar
  21. 21.
    Kamatou GP, Viljoen A (2010) A review of the application and pharmacological properties of α-bisabolol and α-bisabolol-rich oils. J Am Oil Chem Soc 87:1–7CrossRefGoogle Scholar
  22. 22.
    Kim S, Lee J, Jung E, Huh S, Park J-O, J-w Lee, Byun SY, Park D (2008) Mechanisms of depigmentation by α-bisabolol. J Dermatol Sci 52:219–222CrossRefGoogle Scholar
  23. 23.
    Hakozaki T, Takiwaki H, Miyamoto K, Sato Y, Arase S (2006) Ultrasound enhanced skin-lightening effect of vitamin C and niacinamide. Skin Res Technol 12:105–113CrossRefGoogle Scholar
  24. 24.
    Manela-Azulay M, Bagatin E (2009) Cosmeceuticals vitamins. Clin Dermatol 27:469–474CrossRefGoogle Scholar
  25. 25.
    Almeida MM, Alves JMP, Patto DCS, Lima CRRC, Quenca-Guillen JS, Santoro MIRM, Kedor-Hackmann ERM (2009) Determination of tocopheryl acetate and ascorbyl tetraisopalmitate in cosmetic formulations by HPLC. Int J Cosmet Sci 31:445–450CrossRefGoogle Scholar
  26. 26.
    Maia Campos PMBG, Gonçalves GMS, Gaspar LR (2008) In vitro antioxidant activity and in vivo efficacy of topical formulations containing vitamin C and its derivatives studied by non-invasive methods. Skin Res Technol 14:376–380CrossRefGoogle Scholar
  27. 27.
    Gaspar LR, Campos PMBGM (2007) Photostability and efficacy studies of topical formulations containing UV-filters combination and vitamins A, C and E. Int J Pharm 343:181–189CrossRefGoogle Scholar
  28. 28.
    Pedro AS, Detoni C, Ferreira D, Cabral-Albuquerque E, Sarmento B (2009) Validation of a high-performance liquid chromatography method for the determination of (−)-α-bisabolol from particulate systems. Biomed Chromatogr 23:966–972CrossRefGoogle Scholar
  29. 29.
    Perbellini L, Gottardo R, Caprini A, Bortolotti F, Mariotto S, Tagliaro F (2004) Determination of alpha-bisabolol in human blood by micro-HPLC–ion trap MS and head space-GC–MS methods. J Chromatogr B 812:373–377CrossRefGoogle Scholar
  30. 30.
    Scalia S, Giuffreda L, Pallado P (1999) Analytical and preparative supercritical fluid extraction of chamomile flowers and its comparison with conventional methods. J Pharm Biomed Anal 21:549–558CrossRefGoogle Scholar
  31. 31.
    Weber B, Herrmann M, Hartmann B, Joppe H, Schmidt CO, Bertram H-J (2008) HPLC/MS and HPLC/NMR as hyphenated techniques for accelerated characterization of the main constituents in Chamomile (Chamomilla recutita [L.] Rauschert). Eur Food Res Technol 226:755–760CrossRefGoogle Scholar
  32. 32.
    Aoki F, Nakagawa K, Tanaka A, Matsuzaki K, Arai N, Mae T (2005) Determination of glabridin in human plasma by solid-phase extraction and LC-MS/MS. J Chromatogr B Analyt Technol Biomed Life Sci 828:70–74CrossRefGoogle Scholar
  33. 33.
    US Pharmacopeia (2009) General Chapters <1225> Validation of compendial methods. United States Pharmacopeal Convention, Inc., Rockville, Maryland, USAGoogle Scholar
  34. 34.
    Karageorgou E, Samanidou V (2014) Youden test application in robustness assays during method validation. J Chromatogr A 1353:131–139CrossRefGoogle Scholar
  35. 35.
    Commission Decision 2002/657/EC implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Off J Eur Commun L221 8Google Scholar
  36. 36.
    Scortichini G, Annunziata L, Haouet MN, Benedetti F, Krusteva I, Galarini R (2005) ELISA qualitative screening of chloramphenicol in muscle, eggs, honey and milk: method validation according to the Commission Decision 2002/657/EC criteria. Anal Chim Acta 535:43–48CrossRefGoogle Scholar
  37. 37.
    Furman WB, Dorsey JG, Snyder LR (1998) System suitability tests in regulatory liquid and gas chromatographic methods: adjustments versus modifications. Pharm Technol 22:58–65Google Scholar
  38. 38.
    US Food and Drug Administration (1994) Reviewer Guidance, validation of chromatographic methods. Centre for Drug Evaluation and Research, WashingtonGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Jong-Sup Jeon
    • 1
    Email author
  • Han-Taek Kim
    • 1
  • Myeong-Gil Kim
    • 1
  • Moon-Seog Oh
    • 1
  • Se-Ra Hong
    • 1
  • Mi-Hye Yoon
    • 1
  • Ho-Chul Shin
    • 2
  • Jae-Han Shim
    • 3
  • Nehal Aly Afifi
    • 4
  • Ahmet Hacımüftüoğlu
    • 5
  • A. M. Abd El-Aty
    • 2
    • 4
    Email author
  1. 1.Public Health Research DivisionGyeonggi Province Institute of Health and EnvironmentSuwon-SiRepublic of Korea
  2. 2.Department of Veterinary Pharmacology and Toxicology, College of Veterinary MedicineKonkuk UniversitySeoulRepublic of Korea
  3. 3.Biotechnology Research InstituteCollege of Agriculture and Life Sciences, Chonnam National UniversityGwangjuRepublic of Korea
  4. 4.Department of Pharmacology, Faculty of Veterinary MedicineCairo UniversityGizaEgypt
  5. 5.Department of Medical Pharmacology, Medical FacultyAtaturk UniversityErzurumTurkey

Personalised recommendations