A Simple Spectrophotometric Test to Identify Skin Sensitizers and Non-Sensitizers in Chemico (분광광도법을 이용한 간단한 in Chemico 피부감작성 시험법의 개발)

Dong Ho Cha (차동호); 네팔마헤시; Geon Ho Kim (김건호); Kang， Mi Jeong (강미정); Jeong, Tae Cheon (정태천)

doi:10.23032/jaae.2019.13.1.005

A Simple Spectrophotometric Test to Identify Skin Sensitizers and Non-Sensitizers in Chemico

Journal of Alternatives to Animal Experiments
Abbr : J Altern Anim Exp
2019, 13(1), pp.39-47
DOI : 10.23032/jaae.2019.13.1.005
Publisher : Korean Society for Alternatives to Animal Experiments
Research Area : Medicine and Pharmacy > Veterinary > Basic Veterinary Medicine > Laboratory Animal Medicine
Received : November 11, 2019
Accepted : December 13, 2019
Published : December 30, 2019

Dong Ho Cha ¹, 네팔마헤시 ¹, Geon Ho Kim ¹, Kang， Mi Jeong ¹, Jeong, Tae Cheon ¹

¹영남대학교

Candidate

ABSTRACT

For the key event No. 1 in skin sensitization, a hapten should react with endogenous proteins for the initiation of immune reaction. Although Direct Peptide Reactivity Assay (DPRA) has been developed as an alternative method for key event 1, artificially synthesized peptides and HPLC should be employed in performance, which are somewhat inconvenient. In the present study, a simple spectrophotometrical method to identify skin sensitizers in chemico was developed by using cysteamine, an endogenous substance that contains amino and thiol groups. To quantitate remaining cysteamine following the reaction with skin sensitizers, 5,5’-dithiobis-(2-nitrobenzoic acid) (DTNB) was selected for the derivatization. The conditions optimized included: incubation time of cysteamine with test substances, molar ratios of cysteamine to test substances, stability of derivatized products of test substances with optimal concentration and incubation time of DTNB, and spectrophotometrical characteristics of the derivatized product. With the optimized conditions, 11 skin sensitizers and non-sensitizers were tested to know whether the method is feasible or not. Although further optimization is required, it would be a useful screening tool for determining skin sensitization potential of small molecules, because the present method employs simple endogenous cysteamine as an acceptor for sensitizers with a spectrophotometric detection system.

KEYWORDS

skin sensitization, cysteamine, DPRA, spectrophotometry, in chemico

Citation status

* References for papers published after 2023 are currently being built.

[journal] Ahlfors SR / 2003 / Reactivity of contact allergenic haptens to amino acid residues in a model carrier peptide, and characterization of formed peptide-hapten adducts / Skin Pharmacology and Physiology 16 : 59~68

[journal] Anderson SE / 2011 / The LLNA : A brief review of recent advances and limitations / Journal of Allergy 2011 : 1~10

[journal] Autelitano A / 2017 / The reaction of cinnamaldehyde and cinnam(o)yl derivatives with thiols / Acta Pharmaceutica Sinica B 7 : 523~526

[journal] Basketter DA / 2000 / Use of the local lymph node assay for the estimation of relative contact allergenic potency / Contact Dermatitis 42 : 344~348

[journal] Basketter DA / 1999 / A comparison of statistical approaches to the derivation of EC3 values from local lymph node assay dose responses / Journal of Applied Toxicology 19 : 261~266

[journal] Basketter DA / 1992 / Comparison of the local lymph node assay with the guinea-pig maximization test for the detection of a range of contact allergens / Food and Chemical Toxicology 30 : 65~69

[journal] Cho SA / 2019 / High-throughput screening (HTS)-based spectrophotometric direct peptide reactivity assay (Spectro-DPRA) to predict human skin sensitization potential / Toxicology Letters 314 : 27~36

[journal] Cockshott A / 2006 / The local lymph node assay in practice: a current regulatory perspective / Human & Experimental Toxicology 25 : 387~394

[journal] Dimitrov S / 2016 / Accounting for data variability, a key factor in in vivo/in vitro relationships: application to the skin sensitization potency (in vivo LLNA versus in vitro DPRA) example / Journal of Applied Toxicology 36 : 1568~1578

[journal] Gerberick GF / 2004 / Development of a peptide reactivity assay for screening contact allergens / Toxicological Sciences 81 : 332~343

[journal] Gerberick GF / 2007 / Local lymph node assay (LLNA) for detection of sensitization capacity of chemicals / Methods 41 : 54~60

[journal] Gerberick GF / 2007 / Quantification of chemical peptide reactivity for screening contact allergens: A classification tree model approach / Toxicological Sciences 97 : 417~427

[journal] Gerberick GF / 2009 / Investigation of peptide reactivity of pro-hapten skin sensitizers using a peroxidase-peroxide oxidation system / Toxicological Sciences 112(1) : 164~174

[journal] Jowsey IR / 2006 / A future approach to measuring relative skin sensitising potency: a proposal / Journal of Applied Toxicology 26 : 341~350

[journal] Kimber I / 2002 / The local lymph node assay: past, present and future / Contact Dermatitis 47 : 315~328

[journal] Kimber I / 2000 / Cytokines and chemokines in the initiation and regulation of epidermal Langerhans cell mobilization / British Journal of Dermatology 142 : 401~412

[journal] Kimber I / 2010 / Assessment of the skin sensitization potential of topical medicaments using the local lymph node assay: An interlaboratory evaluation / Journal of Toxicology and Environmental Health 53 : 563~579

[journal] Lepoittevin JP / 1991 / Allergic contact dermatitis caused by naturally occurring quinones / Pharmaceutisch Weekblad 13(3) : 119~122

[journal] Nepal MR / 2019 / Identification of pre- and pro-haptens with a β-galactosidase-expressing E. coli cell culture system for skin sensitization / Toxicology Letters 305 : 81~93

[journal] Nepal MR / 2018 / A simple in chemico method for testing skin sensitizing potential of chemicals using small endogenous molecules / Toxicology Letters 289 : 75~85

[journal] Patlewicz G / 2016 / Can currently available non-animal methods detect pre- and pro-haptens relevant for skin sensitization? / Regulatory Toxicology and Pharmacology 82 : 147~155

[journal] Ryan CA / 2001 / Approaches for the development of cell-based in vitro methods for contact sensitization / Toxicology in Vitro 15 : 43~55

[journal] Ryan CA / 2005 / Interactions of contact allergens with dendritic cells: Opportunities and challenges for the development of novel approaches to hazard assessment / Toxicological Sciences 88 : 4~11

[journal] Urbisch D / 2016 / Peptide reactivity associated with skin sensitization: The QSAR Toolbox and TIMES compared to the DPRA / Toxicology in Vitro 34 : 194~203

This paper was written with support from the National Research Foundation of Korea.

KJCKorea
Journal Central

Journal of Alternatives to Animal Experiments 2023 KCI Impact Factor : 0.07

A Simple Spectrophotometric Test to Identify Skin Sensitizers and Non-Sensitizers in Chemico

ABSTRACT

KEYWORDS

Citation status

* References for papers published after 2023 are currently being built.

Journal of Alternatives to Animal Experiments 2023 KCI Impact Factor : 0.07

A Simple Spectrophotometric Test to Identify Skin Sensitizers and Non-Sensitizers in Chemico

ABSTRACT

KEYWORDS

Statistics

Tools

Issue List

Citation status

KCI Citation Counts (0)

REFERENCES (24) * References for papers published after 2023 are currently being built.

Search PDF

Citation

* References for papers published after 2023 are currently being built.