Detail publikace

Colorimetric enzyme-coupled assay for hyaluronic acid determination in complex samples

PEPELIAEV, S. HRUDÍKOVÁ, R. SMIRNOU, D. ČERNÝ, Z.

Originální název

Colorimetric enzyme-coupled assay for hyaluronic acid determination in complex samples

Typ

článek v časopise ve Web of Science, Jimp

Jazyk

angličtina

Originální abstrakt

The present study describes the development of a fast, affordable and reliable method for hyaluronic acid detection in complex samples. The method involves three principle steps. The first is the separation of hyaluronic acid (HA) from interfering glycosaminoglycans as well as mono- and oligosaccharides by cetyltrimethylammonium bromide fractioning. The second is subsequent digestion of HA with Streptococcus pneumoniae hyaluronate lyase to 4,5-unsaturated disaccharides (Delta HA2). The third is the reaction of Delta HA2 with 3-methyl-2-benothiazolinonehydrazone (MBTH) resulting in an intense blue-colored product. The extinction coefficient of Delta HA2-MBTH product is 34,735 mol(-1) at 654 nm. The theoretical sensitivity of the assay is 0.07-0.09 mg/1 HA. The practical sensitivity is 0.3 mg/1; the highest repeatability was achieved in the range of 3-2000 mg/1 HA (r(2) = 0.9994). The analysis took 25-60 min depending on sample complexity. The described method was evaluated in an industrial setting for online monitoring of HA losses during downstream processes and for HA determination in veterinary products. It was positively rated by users and was introduced to routine laboratory practices.

Klíčová slova

Hyaluronic acid assay; Colorimetry; Glycosaminoglycan precipitation; Streptococcus pneumoniae hyaluronan lyase

Autoři

PEPELIAEV, S.; HRUDÍKOVÁ, R.; SMIRNOU, D.; ČERNÝ, Z.

Vydáno

14. 9. 2017

Nakladatel

PERGAMON-ELSEVIER SCIENCE LTD

Místo

OXFORD

ISSN

0014-3057

Periodikum

European Polymer Journal

Ročník

94

Číslo

94

Stát

Spojené království Velké Británie a Severního Irska

Strany od

460

Strany do

470

Strany počet

11

URL

https://www.sciencedirect.com/science/article/abs/pii/S0014305717300551

BibTex

@article{BUT165012,
  author="Stanislav {Pepeliaev} and Radka {Hrudíková} and Dzianis {Smirnou} and Zbyněk {Černý}",
  title="Colorimetric enzyme-coupled assay for hyaluronic acid determination in complex samples",
  journal="European Polymer Journal",
  year="2017",
  volume="94",
  number="94",
  pages="460--470",
  doi="10.1016/j.eurpolymj.2017.07.036",
  issn="0014-3057",
  url="https://www.sciencedirect.com/science/article/abs/pii/S0014305717300551"
}