Quantitative 4-hexylresorcinol determination in muscle tissue of broiler chickens receiving this compound in feeding diet and the influence of its residual concentrations on the microbiological characteristics of poultry products

Cover Page

Cite item

Full Text

Open Access Open Access
Restricted Access Access granted
Restricted Access Subscription or Fee Access

Abstract

Introduction. Plant-derived compounds with antimicrobial activity (phytobiotics) are a relevant alternative to traditional antibiotics in livestock and poultry farming. One of the promising phytobiotics are alkylresorcinols - a large group of phenolic lipids, among which 4-hexylresorcinol has the most pronounced antimicrobial activity.

Goal. To assess the possibility of 4-hexylresorcinol accumulation in muscle tissue of broiler chickens that received this compound as an additive to the basic diet, with an evaluation of the effect of its residual concentrations on microbiological indicators of meat quality.

Material and Methods. Chemically synthesized 4-hexylresorcinol (CAS 136-77-6) with a purity of 98% was used in the study. Experimental groups of Arbor Acres broiler chickens received this compound as an additive to the basic diet for 35 days at doses of 62.5, 125 and 250 mg per 1 kg of feed. The quantitative determination of 4-hexylresorcinol in the muscle tissue of broiler chickens was carried out by high-performance chromatography with triple-quadrupole mass spectrometric detection on a liquid chromatograph-mass spectrometer LCMS-8050 (Shimadzu Corporation, Japan). Microbial contamination of muscle tissue immediately after slaughter and after 5 days of storage at +4°C was analyzed in accordance with national quality standards.

Results. Validation of an HPLC-MS/MS method for the quantitative determination of 4-hexylresorcinol in broiler chicken muscle tissue established a linear range of 50 - 1000 ng/g with deviations no more than ±15% between the nominal and experimentally detected concentrations of this compound. Analysis of biosamples from broiler chickens fed with 4-hexylresorcinol supplement showed the possibility of its accumulation in the range from 50 to 200 ng per 1 gram of muscle tissue with maximum values of 124.4-194.5 ng/g in the experimental group receiving this compound at a dose of 125 mg per 1 kg of feed. The presence of 4-hexylresorcinol in broiler chicken muscles resulted in suppression of mesophilic aerobic and facultative-anaerobic microorganisms in post-slaughter products, which was most pronounced in samples with the maximum accumulation of this compound.

Conclusion. The study results indicate the possibility of 4-hexylresorcinol accumulation in poultry products due to its use as a feed supplement. This observation corresponds to the inclusion of 4-hexylresorcinol in the EAEU Customs Union list of chemical compounds permitted for use in the food industry, but requires further study of its migration along food chains, taking into account the multiple variants of biological activity of alkylresorcinols.

Full Text

Restricted Access

About the authors

D. G. Deryabin

FGBNU «Federal Scientific Center of Biological Systems and Agrotechnologies of the Russian Academy of Sciences»

Author for correspondence.
Email: dgderyabin@yandex.ru
ORCID iD: 0000-0002-9578-7947

Dr.Sc. (Med.), Professor, Head of Laboratory

 

Russian Federation, 29, 9 Janvarya St., Orenburg, 460000

G. K. Duskaev

FGBNU «Federal Scientific Center of Biological Systems and Agrotechnologies of the Russian Academy of Sciences»

Email: gduskaev@mail.ru
ORCID iD: 0000-0002-9015-8367

Dr.Sc. (Biol.), Chief Research Scientist

Russian Federation, 29, 9 Janvarya St., Orenburg, 460000

E. N. Goncharova

LLC SIE «Academy of Innovations»

Email: goncharova.e@vetmag.ru
ORCID iD: 0000-0002-2207-3257

Ph.D. (Chem.), Head of Laboratory

Russian Federation, Talalikhina str. 33, build. 5, Moscow, 125080

T. A. Klimova

FGBNU «Federal Scientific Center of Biological Systems and Agrotechnologies of the Russian Academy of Sciences»

Email: klimovat91@mail.ru
ORCID iD: 0000-0003-4298-1663

Research Scientist

Russian Federation, 29, 9 Janvarya St., Orenburg, 460000

K. S. Lazebnik

FGBNU «Federal Scientific Center of Biological Systems and Agrotechnologies of the Russian Academy of Sciences»

Email: christinakondrashova94@yandex.ru
ORCID iD: 0000-0003-4907-9656

Junior Research Scientist

Russian Federation, 29, 9 Janvarya St., Orenburg, 460000

K. S. Inchagova

FGBNU «Federal Scientific Center of Biological Systems and Agrotechnologies of the Russian Academy of Sciences»

Email: ksenia.inchagova@mail.ru
ORCID iD: 0000-0003-1111-8293

Ph.D. (Biol.), Research Scientist

Russian Federation, 29, 9 Janvarya St., Orenburg, 460000

E. A. Bukareva

FGBNU «Federal Scientific Center of Biological Systems and Agrotechnologies of the Russian Academy of Sciences»

Email: elenka_rs@mail.ru
ORCID iD: 0000-0002-1622-1284

Ph.D. (Biol.), Acting Head of Laboratory

Russian Federation, 29, 9 Janvarya St., Orenburg, 460000

I. E. Laryushina

FGBNU «Federal Scientific Center of Biological Systems and Agrotechnologies of the Russian Academy of Sciences»

Email: inhip@mail.ru

Ph.D. (Med.), Senior Research Scientis

Russian Federation, 29, 9 Janvarya St., Orenburg, 460000

References

  1. Тимофеев Н.П. Фитобиотики в мировой практике: виды растений и действующие вещества, эффективность и ограничения, перспективы (обзор). Аграрная наука Евро-Северо-Востока. 2021;22(6):804–825. [Timofeev N.P. Phytobiotics in world practice: plant species and active substances, efficiency and limitations, perspectives (review). Agricultural Science Euro-North-East. 2021;22(6):804-825. (In Russ.)]. doi: 10.30766/2072-9081.2021.22.6.804-825.
  2. Подобед Л. Фитобиотики в кормлении животных. Животноводство России. 2018;7:57–58. [Podobed L. Phytobiotics in animal feeding. Animal husbandry of Russia. 2018;7:57–58. (In Russ.)].
  3. Kozubek A., Tyman J.H. Resorcinolic Lipids, the Natural Non-isoprenoid Phenolic Amphiphiles and Their Biological Activity. Chemical Reviews. 1999;99(1):1–26. doi: 10.1021/cr970464o.
  4. Zarnowski R., Kozubek А. Alkylresorcinol Homologs in Pisum sativum L. Varieties. Zeitschrift für Naturforschung C. 1999;54(1-2):44–48. doi: 10.1515/znc-1999-1-208.
  5. Żarnowska Е.D., Żarnowski R., Kozubek A. Alkylresorcinols in Fruit Pulp and Leaves of Ginkgo biloba L. Zeitschrift für Naturforschung C. 2000;55(11-12):881–885. doi: 10.1515/znc-2000-11-1206.
  6. Knödler M., Reisenhauer K., Schieber A. et al. Quantitative determination of allergenic 5-alk(en)ylresorcinols in mango (Mangifera indica L.) peel, pulp, and fruit products by high-performance liquid chromatography. Journal of Agricultural and Food Chemistry. 2009;57(9):3639–3644. doi: 10.1021/jf803934p.
  7. Deryabin D.G., Tolmacheva, A.A. Antibacterial and Anti-Quorum Sensing Molecular Composition Derived from Quercus cortex (Oak bark) Extract. Molecules. 2015;20(9):17093–17108. doi: 10.3390/molecules200917093.
  8. Nikolaev Y.A., Tutel'yan A.V., Loiko N.G. et al. The use of 4-Hexylresorcinol as antibiotic adjuvant. PLoS One. 2020;15(9):e0239147. doi: 10.1371/journal.pone.0239147.
  9. Инчагова К.С., Русакова Е.А. Исследование регуляторных эффектов малых молекул растительного происхождения в отношении бактериальных биосенсоров с различными системами «quorum sensing» LuxI/LuxR-типа. Вопросы биологической, медицинской и фармацевтической химии. 2023;26(11):40−46. [Inchagova K.S., Rusakova E.A. Investigation of the regulatory effects of small molecules of plant origin in relation to bacterial biosensors with various LUXI/LuxR-type "quorum sensing" systems. Problems of Biological, Medical and Pharmaceutical Chemistry. 2023;26(11):40-46. (In Russ.)]. doi: 10.29296/25877313-2023-11-07.
  10. Дерябин Д. Г., Камаева А. А., Толмачева А. А. и др. Влияние алкилоксибензолов на индуцируемые гомосеринлактонами проявления кворум сенсинга у бактерий. Прикладная биохимия и микробиология. 2014;50(4):391-397. [Deryabin D.G., Kamayeva A.A., Tolmacheva A.A. et al. The effects of alkylhydroxybenzenes on homoserine lactone-induced manifestations of quorum sensing in bacteria. Applied Biochemistry and Microbiology. 2014;50(4):391-397. (In Russ.)]. doi: 10.7868/S0555109914040199.
  11. Zabolotneva A.A., Gaponov A.M., Roumiantsev S.A. et al. Alkylresorcinols as New Modulators of the Metabolic Activity of the Gut Microbiota. International Journal of Molecular Sciences. 2023;24(18):14206. doi: 10.3390/ijms241814206.
  12. Iyengar R., Bohmont C.W., Mcevily A.J. 4-Hexylresorcinol and prevention of shrimp blackspot: Residual analyses. Journal of Food Composition and Analysis. 1991;4(2):148-157. doi: 10.1016/0889-1575(91)90008-t.
  13. Zavyalov O., Duskaev G., Kurilkina M. Effect of feeding bioactive compounds identified from plant extracts (4-hexylresorcinol, 7-hydroxycoumarin, and gamma-octalactone) on the productivity and quality of broiler meat. Veterinary World. 2022;15(12): 2986-2996. doi: 10.14202/vetworld.2022.2986-2996.
  14. Шестопалов А.В., Гапонов А.М., Заболотнева А.А. и др. Алкилрезорцинолы – новые потенциальные биорегуляторы в системе суперорганизма (человек-микробиота). Известия Российской академии наук. Серия биологическая. 2022;3:246-257. [Shestopalov A.V., Gaponov A.M., Zabolotneva A.A. et al. Alkylresorcinols: new potential bioregulators in the superorganism system (human–microbiota). Biology Bulletin. 2022;3:246-257. (In Russ.)]. doi: 10.31857/S1026347022030155.
  15. Технический регламент Таможенного союза ТР ТС 029/2012 «Требования безопасности пищевых добавок, ароматизаторов и технологических вспомогательных средств» (с изменениями на 18 сентября 2014 года). [Technical Regulation of the Customs Union TR TS 029/2012 “Safety requirements for food additives, flavorings and technological aids” (as amended on September 18, 2014). (In Russ.)]; https://ucsol.ru/images/docs/TR/029_2012_tr_ts.pdf.
  16. ГОСТ 10444.15-94. Продукты пищевые. Методы определения количества мезофильных аэробных и факультативно-анаэробных микроорганизмов. [GOST 10444.15-94. Food products. Methods for determining the number of mesophilic aerobic and facultative-anaerobic microorganisms. (In Russ.)]; https://internet-law.ru/gosts/gost/18812/.
  17. ГОСТ 31747-2012. Продукты пищевые. Методы выявления и определения количества бактерий группы кишечных палочек (колиформных бактерий). [GOST 31747-2012. Food products. Methods of detection and determination of the number of Escherichia coli bacteria (coliform bacteria) (In Russ.)]; https://internet-law.ru/gosts/gost/52791/.
  18. ГОСТ 31468-2012. Мясо птицы, субпродукты и полуфабрикаты из мяса птицы. Метод выявления сальмонелл. [GOST 31468-2012 Poultry meat, by-products and semi-finished products from poultry meat. Method for the detection of salmonellae (In Russ.)]; https://internet-law.ru/gosts/gost/52573.
  19. VICH topic GL49: Studies to evaluate the metabolism and residues kinetics of veterinary drugs in human food-producing animals: validation of analytical methods used in residue depletion studies; https://www.ema.europa.eu/en/documents/scientific-guideline/vich-gl49-studies-evaluate-metabolism-and-residue-kinetics-veterinary-drugs-food-producing-animals-validation-analytical-methods-used-residue-depletion-studies_en.pdf.
  20. Hao Y., Gao X., Xia W. Determination of 4-Hexylresorcinol in Shrimp Samples by Solid Phase Extraction Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry. Molecules. 2018;23(9):2173. doi: 10.3390/molecules23092173.
  21. Guandalini E., Ioppolo A., Mantovani A., Stacchini P., Giovannini C. 4-Hexylresorcinol as inhibitor of shrimp melanosis: efficacy and residues studies; evaluation of possible toxic effect in a human intestinal in vitro model (Caco-2); preliminary safety assessment. Food Additives and Contaminants. 1998;15(2):171–180. doi: 10.1080/02652039809374627.
  22. NTP Toxicology and Carcinogenesis Studies of 4-Hexylre-sorcinol (CAS No. 136-77-6) in F344/N Rats and B6C3F1 Mice (Gavage Studies). Natl Toxicol Program Tech Rep Ser. 1988 May:330:1–166; https://pubmed.ncbi.nlm.nih.gov/12732906/.
  23. Жаринов А.И., Кузнецова О.В., Жукова А.Ю., Костенко Ю.Г. К вопросу контаминации мяса птицы бактериями Serratia marcescens. Мясные технологии. 2019;4(196):56-59. [Zharinov A.I., Kuznetsova O.V., Zhukova A.Y., Kostenko Y.G. To the question of contamination of poultry meat with Serratia marcescens bacteria. Meat Technologies. 2019;4(196):56–59. (In Russ.)]. doi: 10.33465/2308-2941-2019-4-56-58.
  24. Сейтжанова М.Е. Оценка качества различных видов мясного сырья и готовой продукции. European Student Scientific Journal. 2018; 3:12–12. [Seitzhanova M.E. Quality assessment of different types of meat raw materials and finished products. European Student Scientific Journal. 2018; 3:12–12. (In Russ.)]; https://sjes.esrae.ru/en/article/view?id=433.

Supplementary files

Supplementary Files
Action
1. JATS XML
Download
2. Fig. 1: Structural formula of 4-hexylresorcinol

Download (5KB)
Indexing metadata
3. Fig. 2. HPLC-MS/MS chromatogram of the determination of 4-hexylresorcinol at the NPCO level (50 ng/g). Top: abscissa axis – retention time on the column, min; ordinate axis – signal intensity, imp. Bottom: parameters of its mass spectrometric detection

Download (116KB)
Indexing metadata
4. Fig. 3. Graduation relationship for the quantitative determination of 4-hexylresorcinol in broiler chicken muscle tissue based on three parallel measurements of one analytical cycle. On the abscissa axis [4-GR] is the nominal (contributed) concentration of hexylresorcinol (ng/g); on the ordinate axis [S] is the area of the chromatographic peak (S) during HPLC-MS/MS determination

Download (40KB)
Indexing metadata
5. Fig. 4. Correspondences between the presence of residual amounts of 4-hexylresorcinol (along the abscissa axis, ng/g) and the microbial contamination of muscle tissue of broiler chickens, characterized by the QMAFAnM indicator (along the ordinate axis, CFU/g) immediately after slaughter (white diamonds) and after 5 days of storage at +4 °C (black diamonds). The standard value of bacterial contamination according to GOST 10444.15-94 [16] is shown by the horizontal line.

Download (42KB)
Indexing metadata

Copyright (c) 2024 Russkiy Vrach Publishing House