Ïóáëèêàöèè â “Nauka i studia”( Przemysl, Ïîëüøà) èëè â ×åõèè

Ñåêöèÿ “Áèîëîãè÷åñêèå íàóêè/Áèîõèìèÿ è áèîôèçèêà” èëè

“Ìåäèöèíà/Ýêñïåðèìåíòàëüíàÿ ôàðìàêîëîãèÿ”èëè

“Õèìèÿ/Õèìèêî-ôàðìàöåâòè÷åñêîå ïðîèçâîäñòâî”

 

Mamina Î.Î., Kovalenko Z.I.

Ukrainian National University of Pharmacy,t.Kharkov

Identification of synthetic food dye Allura Red by HPLC

 

Allura Red, disodium 2-hydroxy-1-(2-methoxy-5-methyl-4-sulfonato- phenylazo)-naphtalene-6-sulfonate is synthetic food dye, E 129 according to the international enumeration system of food additives [1].

Allura Red widely used in food, pharmaceutical and cosmetic industries, but chronic use causes allergies, disorders of biochemical reactions and cancer [1,2]. This dye affect the chromosomes and violates the division of cells in the tissues [2].

To determine the synthetic food dyes uses such analytical methods, as chromatographic (HPLC, TLC, electrophoresis) and spectral (UV and visible spectrophotometry)[1,3]. HPLC-methods are characterized by various chromatograph conditions, depending on individual properties of food dyes. The purpose of work - the identification of Allura Red when using unified conditions HPLC suitable for studies of pharmaceuticals, food products, biological objects.

For research standards have been used  «Dyes  analytical  standart kit 10 pcs», Institute of  dyes and organic products (IBPO),  Poland. 

The chromatographic process of Allura Red has been conducted  on microcolumn liquid chromatograph «Milichrom À-02»  on base research union «Ànalitika» in reversed-phase variant with using of metalic column with non-polar absorbent Prontosil 120-5C 18 AQ, 5 μm.

Allura Red has been eluted with mobile phase in the mode of linear gradient – from eluent À (5 % acetonitrile and 95% buffer solution - 0,2 Ì solution of lithium perchlorate in 0,005 Ì solution perchloric acid) to eluent B (100% acetonitrile) as during 40 min. Regeneration of column has been conducted  during 2 min with mixture of solvents.The flow rate of the mobile phase has been formed 100 μl/min,  injection volum –  1  μl. The detection of of Allura Red has been conducted by UV- detector at 7 wavelengths: 210, 220,  240, 250, 260, 280, 350 nm. The optimal values of column temperature –  40°Ñ and pressure of pump – 4,2 MPa.

mV
 
The symmetry factor (As) of peaks (1,52 ± 0,03) have been received in result of the chromatographic process of Allura Red with using unified conditions HPLC. The identification of of Allura Red has been conducted with using absolute parameters of retention time (tR) and retention volum (VR), and spectral relations, which  determined  as ratio of values of absorbance at wavelengths - 220-350 nm – to values of absorbance at  210 nm (Fig.,Tab.).                                                                

Time¸ min
 

Fig. Chromatogram of Allura Red (aqueous solution concentration 8,5 mg / ml)

 

Table

Retention parameters and spectral relations (n = 5)

 

 

 

Allura Red

Retention parameters

Factor of capacity, k´

tR, min

VR, μl

8,21 ± 0,02

821,1

4,47

Spectral relations

À220 nm

À 210 nm

À240 nm

À210 nm

À250 nm

À 210 nm

À260 nm

À 210 nm

À280 nm

À 210 nm

À350 nm

À 210 nm

0,915

0,761

0,567

0,386

0,265

0,072

 

Results of research can be recommended for the analysis of pharmaceuticals, food and biological objects on synthetic food dyes.

Conclusions

1. The HPLC-study of Allura Red  with the use of a unified conditions of analysis suitable for studies of pharmaceuticals, food products, biological objects  was conducted.

2. The basic parameters of identification of Allura Red have been defined  in terms of analysis.

Literature

1. Bolotov V.M. Food dyes: classification, properties, analysis, application/ V.M. Bolotov, À.P. Nechaev, L.À.Sarafanova.- SPb: Publishing house GIORD, 2008. -  240 P.

2. Pesnya D. S. Study of toxic and genotoxic effects of synthetic food dyes method Allium test / D. S. Pesnya, A. S. Romanovsky, I. M. Prokhorov. // The Yaroslavl pedagogical Bulletin - 2012 - ¹ 3 – Vol. III (Natural Sciences)- pp.86-93.

3. Kiseleva M. G.  Optimization of Conditions for the HPLC Determination of Synthetic Dyes in Food / M. G. Kiseleva, V.V. Pimenova, K. I. Eller//J. of Anal. Chem. – 2003. - Vol. 58, No. 7. - pp. 685–690.