WO2020128774A1

WO2020128774A1 - Development of boron derivative chromotographic filling material

Info

Publication number: WO2020128774A1
Application number: PCT/IB2019/060829
Authority: WO
Inventors: Erdal ERTAŞ; Öznur KARAOĞLU
Original assignee: Scientific and Technological Research Council of Turkey TUBITAK
Current assignee: Scientific and Technological Research Council of Turkey TUBITAK
Priority date: 2018-12-19
Filing date: 2019-12-16
Publication date: 2020-06-25
Anticipated expiration: 2021-06-19
Also published as: TR201819830A2

Abstract

In the context of the present invention, the new boron derived zeolitic adsorbent, which are named as DAZB and DSZB were obtained from the chemical reactions between various equivalents of boric acid and different zeolitic structures bearing free silanol/aluminum hydroxide groups formed after dealumination/desilication process of the corresponding zeolitic structures in different Si/AI ratio. The products with the content of the mixture of sucrose and the colorants named Sunset Yellow and Patent Blue V were processed to be separated to the issued components by using DAZB and DSZB filled solid phase extraction cartridges. The present invention has shown that separation processes, which are similar to the issued colorants and the sugar compounds, can be conducted in a highly inexpensive way by using newly developed DAZB and DSZB adsorbents. Thus, hereby presented DAZB and DSZB filling adsorbents are highly promising materials either in mass production sections or small scale R&D departments of the companies or research laboratories which are interested in food products, chemicals, pharmaceuticals, environmental processes, and related themes.

Description

DEVELOPMENT OF BORON DERIVATIVE CHROMOTOGRAPHIC FILLING

MATERIAL

Field of Invention

The invention relates to the synthesis, characterization and applications of the novel boron-derived zeolite filling materials for chromatographic studies.

The chromatographic filling materials have high potential to be used for purification and recovery studies for industrial food products, pharmaceuticals, and chemical production processes due their easy production, low-cost and abundant natural sources of the related chromatographic materials.

Background Information

Zeolite, dealuminated zeolite and boric acid are widely known materials. There are many applications of such materials in literature [Weitkamp, Jens. "Zeolites and catalysis." Solid State lonicsl 31.1 (2000): 175-188.; Eroshenko, Valentin, et al. "Energetics: a new field of applications for hydrophobic zeolites." Journal of the American Chemical Society 123.33 (2001 ): 8129-8130; Tao, Yousheng, et al. "Mesopore-modified zeolites: preparation, characterization, and applications." Chemical reviews 106.3 (2006): 896-910]

Dealumination and desilication of zeolites in acidic and alkaline conditions have been performed by many groups. Thus, the zeolite modification by these dealumination and desilication processes are well-known studies [Silaghi, Marius-Christian, Celine Chizallet, and Pascal Raybaud. "Challenges on molecular aspects of dealumination and desilication of zeolites." Microporous and Mesoporous Materials 191 (2014): 82- 96.; Verboekend, Danny, and Javier Perez-Ramirez. "Design of hierarchical zeolite catalysts by desilication." Catalysis Science & Technology 1 .6 (201 1 ): 879-890.; Lutz, Wolfgang. "Zeolite Y: Synthesis, modification, and properties-A case revisited." Advances in Materials Science and Engineering (2014)].

Zeolites are often used in catalytic reactions as catalysts or the catalyst components scaffolds which carries the catalytic groups. For example, zeolites are favored to be used in petrochemical industry for applications like cracking as a catalyst [Masters, A. F.; Maschmeyer, T. "Zeolites - From Curiosity to Cornerstone." Microporous Mesoporous Mater 142, (201 1 ), 423-438] On the other hand, adsorption of boric acid on zeolites and investigation of catalytic effects were investigated by some research groups [Zhang, Wengui, et al. "Efficient dehydration of bio-based 2, 3-butanediol to butanone over boric acid modified HZSM-5 zeolites." Green chemistry 14.12 (2012), 3441 -3450.; Zendehdel, Mojgan, and M. Solimannejad. "Interaction between NaY Zeolite and boric Acid: a preliminary computational study." Chemistry of Solid Materials ' l (2013), 57-63.; Asaftei, luliean Vasile, et al. "Conversion of Industrial Feedstock Mainly with Butanes and Butenes Over B-HZSM-5 and Zn-HZSM-5 Modified Catalysts." Revista de Chimie 66.3 (2015): 336-341 ]

Boronation of zeolite was not studied and boron-derived zeolite was not used as chromatographic filling materials. In addition, there are studies in the state of the art to obtain boron esters of silicon-containing compounds and polymeric borosiloxane substances [(O-Si-O-B (OR, H); R: aromatic or aliphatic groups]) and their application. [Yajima, S., Okamura, K., Hayashi, J., & Shishido, T. (1979). U.S. Patent No. 4,152,509. Washington, DC: U.S. Patent and Trademark Office.; Polymeric organosilanol-boronic acid reaction products and method for making same. U.S. Patent No 2,517,945, 1950.; Riccitiello, Salvatore R., Ming-Ta S. Hsu, and Timothy S. Chen. "Boron-containing organosilane polymers and ceramic materials thereof." U.S. Patent No. 4,767,728. 30 Aug. 1988]

The Solution of the Invention to the Technical Problem

A chromatographic filling material (adsorbent) should be a low-cost, easily applicable material, which also should posses good separation properties. In this context, alternative ways of reducing production costs should be sought. Furthermore, it is necessary to develop new materials that can solve the problems of the present analytical techniques and mixtures containing a large number of compounds of the chromatographic filling materials, such as not being able to purify well, purification procedures being long, being unstable at different pH ranges.

There are also many studies about development of chromatographic filling materials which are derivatives of polymeric resins or silica gels in the literature as alternatives to the commercially available and frequently used chromatographic filling materials. [Hosoya, Ken, et al. "Polymer packing material for liquid chromatography and a producing method thereof." U.S. Patent No. 5,990,182. 23 Nov. 1999.; Frechet, Jean MJ, Frantisek Svec, and Ken Hosoya. "Process for producing uniform macroporous polymer beads." U.S. Patent No. 5,130,343. 14 Jul. 1992.; Harris, William I. "Seeded porous copolymers and ion-exchange resins prepared therefrom." U.S. Patent No. 5,231 ,115. 27 Jul. 1993] Octadecyl (C18), octyl (C8), cyano, amine and phenyl, dihydroxypropane (diol) modified silica gels and silica gel 60 are commercially well- known and frequently used filling materials. These filling materials are sold for end- users as powder chromatographic filling materials or coated over glass or aluminum thin layer substrates as modified forms, filled liquid chromatography columns and solid phase extraction (SPE) cartridges. However, the high production costs of those adsorbents reduce the industrial preference of such adsorbents. Therefore, developing absorbents via low-cost and facile synthetic methods from abundant raw materials are needed to reduce the production costs.

The present invention includes the development (syntheses, characterizations, applications and related subjects) of novel chromatographic filling materials that are suitable for low-cost industrial and laboratory scale applications and can be an alternative to commercial available filling materials commonly used in the analytical or chromatographic techniques. The boron-derived zeolites (boric acid aluminates) were developed by a method similar to the production of borosiloxane compounds from silicon-containing structures. Boron-derived zeolites (boron-derivedaluminosilicate) have been developed by attaching boric acid to the free metal hydroxyl (M-OH) groups in the structure of dealumine and desilicated zeolites obtained from various cheap aluminasilicate compounds (zeolites) used as starting products (DAZB: Boron-derived zeolite synthesized with dealumine zeolite; DSZB: Boron-derived zeolite synthesized with desilyl zeolite).

Improved DAZB and DSZB filling materials are completely different from those borosiloxanes, which have been in the state of art. The techniques used in development and applications of boron derivatives filling adsorbents are completely different from the boroxilanes materials production and applications. Developed filling materials are expected to attract high attention in either industry or fundamental research due to their easy and low-cost syntheses from abundant natural sources.

Descriptions of the Figures

Figure 1. Block flow diagram of boron derived zeolite synthesis process using dealuminated zeolite (DAZ)

Figure 2. Block flow diagram of boron derived zeolite synthesis process using desilicated zeolite (DAZ) Figure 3. Schematic illustration of bonding of boric acid to the silanol groups of DAZ and DSZ derivatives

Figure 4. Powder XRD patterns of DSZB DAZB and ZSM-5 structures

Figure 5. FT-IR spectrums of DAZ and DSZ derivatives between the wavenumbers 400 - 1800 cm ¹

Figure 6. NIR spectrums of DAZB and DSZB structures between the wavenumbers 5700 - 7700 cm-¹

Figure 7. NIR spectrums of DAZB and DSZB structures between the wavenumbers 4000 - 6000 cm ¹

Figure 8. NIR spectrums of DAZB and DSZB structures between the wavenumbers 4000 - 7700 cm-¹

Figure 9. FT-IR spectrums of DAZB and DSZB structures between the wavenumbers 450 - 1800 cm ¹ and corresponding chemical bonds

Figure 10. FT-IR spectrum of DAZB between the wavenumbers 450 - 4000 cm ¹ and corresponding chemical bonds

Figure 11. FT-IR spectrum of DSZB between the wavenumbers 450 - 4000 cm ¹ and corresponding chemical bonds

Figure 12. FT-IR spectrum of DAZB and DSZB in various wavenumbers and their corresponding chemical bonds

Figure 13. SEM images of the DAZB (in 10 pm and 30 pm scale bar)

Figure 14. SEM images of the DSZB (in 10 pm and 30 pm scale bar)

Figure 15. Table of surface area (Brunauer-Emmett-Teller, BET), particles size and boron ratios of the DAZB and DSZB adsorbents

Figure 16. Table of FIPLC analysis results of sugars and food colorant recovery with the issued DAZB and DSZB SPE cartridges study

Brief Description

Figure 1.

A. Steps of dealumination reaction under acidic conditions 1. ZSM-5, X or A type zeolite 2. HCI, H2SO4 or HNO3

3.1. Temperature (A)

4. Cooling 5.1. Solution

B. Filtering step

6. Deionized water

7.1. The filtrate C. Washing steps

6. Deionized water

8. Removing acid solutions

9.1. Washed dealuminated zeolite

D. Drying step

10. Temperature (D)

11.1. Dried dealuminated zeolite

E. Cooling step

12. Vacuum

4. Cooling 13.1. Cooled down dealuminated zeolite

F. Steps of bonding boric acid or boric acid esters (borylation)

14. Boric acid, boric acid methyl alcohol esters, boric acid ethyl alcohol esters

15. Xylene 12. Vacuum 16. Fleating

17.1. Stirring

B. Filtering Step

12. Vacuum

18.1. The filtrate

G. Step of the washing the filtrate with organic solvents 19. Dichloromethane

20.1. The filtrated washed with organic solvent (G)

H. Drying

21. Temperature 22.1. Dried mixture (FI)

I. Cooling

12. Vacuum

4. Cooling

23.1. Boron derived zeolite (DAZB) Figure 2.

J. Steps of desilication reaction under basic conditions 1. ZSM-5, X or A type zeolite

24. NaOH or KOH

3.2. Temperature (A) 4. Cooling

5.2. Solution

B. Filtering step

6. Deionized water

7.2. The filtrate (B) C. Washing step

6. Deionized water

25. Removing basic solutions

9.2. Washed desilicated zeolite D. Drying step

10. Temperature (D) 11.2. Dried desilicated zeolite

E. Cooling step

12. Vacuum

4. Cooling 13.2. Cooled down desilicated zeolite

F. Steps of bonding boric acid or boric acid esters

15. Xylene 12. Vacuum 16. Heating

17.2. Stirring (F)

B. Filtrating step

12. Vacuum

18.2. The filtrate G. Step of the washing the filtrate with organic solvents

19. Dichloromethane

20.2. The filtrated washed with organic solvent (G)

H. Drying

21. Temperature 22.2. Dried mixture (I)

I. Cooling

12. Vacuum

4. Cooling

23.2. Boron derived zeolite (DSZB) Description of Invention

Boric acid and three different zeolitic structures which have different Si/AI ratios, namely ZSM-5((Si/AI >50), X type or A type zeolites (Si/AI<5), are starting raw materialsof the boron derived zeolite filling adsorbents , which are commercially available and cheap . The production steps of the adsorbents by using these starting materials are described in Figure 1 and Figure 2. binding of boric acid to silanol groups of the DAZ and DSZ are also described in Figure 3.

I-) Dealumination of zeolites and improving related boron derivatives zeolites

Dealumination reaction is carried out in the presence of the mineral acid solution (A). 40-70 g of zeolite (ZSM-5, X or A type zeolite) (1 ) is added inside reaction vessel (A).

1 -4 times volume (preferably 3 times) of the zeolites 0.1 -8 N HCI, FI2SO4, or FINO3 (2) is poured slowly to the media, then the reaction is initiated. The reaction is carried out by stirring at 70-180 °C (3.1 ) for 1 -24 hours and cooled down (4) to room temperature afterwards. Thus, acidic solution of dealuminated zeolite (5.1 ) is obtained at the end of the reaction. Filtration (B) of the solid phase is performed after addition of 1 -4 times (preferably 3 times) volume of deionized water (6) and at the end of the this step washed filtrated is obtained (7.1 ). The filtrate (7.1 ) is washed with 1 -4 times volume of the filtrate (preferably 3 times) deionized water (6) to remove acid (8) from the filtrate (7.1 ) and the washing step (C) is repeated 1 -3 times (preferably 2 times). Thus, the mineral acid was used in dealumination steps (A) was removed from the filtrate (checked by using pH meter). Washed dealuminated (DAZ) (9.1 ) is obtained at the end of the filtrating (B) and washing steps (C). DAZ is dried (D) at 110-130 °C (10) during 12 hours. Then, DAZ is cooled down (E) to room temperature (4) under vacuum (12) and at the end of this step cooled DAZ (13.1 ) is obtained as the main product. Boric acid, boric acid ethyl alcohol esters and boric acid methyl alcohol esters are used in the steps of bonding boric acid or boric acid esters (F). 40 -70 g (preferably 50g) of DAZ (13.1 ) and 1 -3 times volume (preferably 2.5 times) of DAZ (13.1 ) xylene (15) are added to reaction vessel. Then 0.05-1 mol ratio boric acid, boric acid ethyl alcohol esters and boric acid methyl alcohol esters (14) are added to reaction media. The reaction mixture is stirred and heated (16) at 170-200 °C (preferably at 180 °C) under vacuum (12). At the end of the reaction, DAZB is formed in reaction mixture (17.1 ) as a major product, then the reaction mixture is filtered (B) under vacuum (12). In order to removed xylene from the filtrate (18.1 ), the filtrate is washed with organic solvents (G) using 1 -3 times (preferably 2 times) volume of the filtrate dichloromethane (19). The filtrated washed with organic solvent (G) is dried (H) by heating (21 ) at 80-120°C (preferably 100 °C) for 12 hours and dried materials (22.1 ) is cooled down (I) to room temperature (4) under vacuum (12). As a result, boron derivate zeolite (23.1 ) is obtained from DAZ at the end of this step.

The chemical and physical properties of the produced materials were investigated by using feasible techniques. The Si/AI ratios, number of Si-OH and AI-OH groups, their crystalline structural sequence, particle size distribution and surface images were collected by using the powder X-ray diffraction (XRD), Inductively Coupled Plasma (ICP-OES), Inductively Coupled Plasma Mass Spectroscopy(ICP-MS), Near Infrared Spectroscopy (NIR), Fourier Transform Infrared Spectroscopy (FT-IR), Scanning Electron Microscopy (SEM).

Considering the results of the analyses and crystallinity patterns of the final compounds, the optimized conditions to yield dealuminated zeolite was found as 2N HCI, temperature as 150 °C and reaction duration as 6 hours.

II-) Desilication of zeolites and improving related boron derivatives zeolites

Desilication reaction is carried out in the presence of alkaline solution (J). 40-70 g of zeolite (ZSM-5, X or A type zeolite) (1 ) is added inside reaction vessel (J). 1 -4 times volume (preferably 3 times) of the zeolites 0.1 - 4N NaOFI, or KOFI (24) is poured slowly to the media. The reaction mixture is stirred at 100-130 °C (preferably 110 °C) (3.2) for 3-12 hours (3.1 ) and cooled down (4) to room temperature afterwards. Thus, alkaline solution including desilicated zeolite (5.2) is obtained at the end of the reaction. Afterward 1 -4 times (preferably 3 times) volume of the desilicated zeolite (5.2) deionized water (6) is added to reaction area and then filtrated (B) to obtained desilicated zeolite filter (7.2).

Alkaline solution is removed (25) from the filter with deionized water (6) using 1 -4 times (preferably 3 times) volume of filter, this washing step (C) is repeated 1 -3 times (preferably 2 times) to provide to washed desilicated zeolite (DSZ) (9.2). Alkaline solution (24), which is added for desilication (J) is entirely removed (checked by using pFH meter). DSZ (11.2) is dried (D) by heating (10) at 110-130^°C for 12 hours. It was cooled down (E) to room temperature (4) under vacuum (12) and then cooled DSZ (13.2) is obtained. Boric acid, boric acid ethyl alcohol esters and boric acid methyl alcohol esters are used in the steps of bonding boric acid or boric acid esters (F). 40-70 g (preferably 50 g) of DSZ (13.2) and xylene (15) which is 1 -3 times by volume (preferably 2.5 times) of DSZ (13.2) are added to reaction vessel. Then 0.05-1 mol ratio boric acid, boric acid ethyl alcohol esters and boric acid methyl alcohol esters (14) are added to reaction media. The reaction mixture is heated (16) at 170-200 °C (preferably at 180 °C) for under vacuum (12). At the end of the reaction, DSZB (13.2) is formed in reaction mixture (17.2) as a major product, then the reaction mixture is filtered (B) under vacuum (12). In order to removed xylene from the filtrate (18.2), the filtrate is washed with organic solvents (G) using 1 -3 times (preferably 2 times) volume of the filtrate dichloromethane (19). The filtrated (20.2) is dried (H) with heating (21 ) at 80-120°C (preferably 100 °C) for 12 hours and dried materials (22.2) is cooled down (I) to room temperature (4) under vacuum (12). As a result, boron derivate zeolite (23.2) is obtained from DSZ at the end of this step. The chemical and physical properties of the produced materials were investigated by using feasible techniques. The Si/AI ratios, number of Si-OH and AI-OH groups, their crystalline structural sequence, particle size distribution and surface images were collected by using the powder X-ray diffraction (pXRD), Inductively Coupled Plasma (ICP-OES), Inductively Coupled Plasma Mass Spectroscopy(ICP-MS), Near Infrared Spectroscopy (NIR), Fourier Transform Infrared Spectroscopy (FT-IR), Scanning Electron Microscopy (SEM).

Considering the results of the analyses and crystallinity patterns of the final compounds, the optimized conditions to yield desilicated zeolite was found as 2N KOFI, 110 °C as temperature and 6 hours for reaction duration.

In Figure 4, XRD patterns of the DAZB and DSZB which were produced from equimolar boric acid treatment of DAZ and DSZ which were obtained from starting material ZSM-5 under optimized conditions can be seen. The XRD results show that the general crystalline structures of zeolites are preserved and the effects of boron groups on the main structure. . The peaks which cannot be observed for DAZ and DSZ at 27.94 and 28.48 2Q degrees appeared after boron modification. Additionally, it was also observed that the peaks at 27.94 and 28.48 2Q degrees of DAZB have higher intensity than the peaks of DSZB at the same region. The important functional groups and their corresponding transmittance wavenumbers of DAZs and DSZs which were obtained in the abovementioned optimized conditions were shown in Figure-5. The intensities are different, this is because DAZ and DSZ contains different Si/AI ratio and different amount of AI-OH and Si-OH groups in the same unit surface. In addition, it is shown that, when DAZ and DSZ are treated with acid or base their particle size is reduced from their pristine zeolite. On the contrary, particle size of the DAZB and DSZB increases after borylation of the DAZ and DSZ.

The NIR spectrums of DAZB and DSZB products which were formed via boric acid treatment of the optimized corresponding DAZ and DSZ precursors in different equivalents (0,1 ; 0,2; and 0,5 boric acid/zeolite, w/w) can be seen on Figure 6 and Figure 7. With the increase of the boric acid amount in the reaction, the changes on intensity of the corresponding peaks were monitored. The increase of the peaks representing Si-O-B and B-OFI and the decrease of the peaks of AI-OH and Si-OH groups after formation of DAZB and DSZB from corresponding DAZ and DSZ precursors were monitored.

In Figure 8, the selected (from Figure 6 and Figure 7) most intense NIR spectrums of DAZB and DSZB materials obtained from DAZ and DSZ by treating them with an equimolar amount of boric acid can be seen. The peaks at 7082 cm ¹ belong to free silanol (Si-OH) groups, and their intensity reduces after boric acid treatment. The peaks at 6573 cm ¹ and 6141 cm ¹ belong to the bonds of B-O-Si groups, which were formed after formation of boronic moieties on the free silanol groups. The intensity of the peaks at 6573 cm ¹ and 6141 cm ¹ increases depending on the molar ratio of boron used in the reaction (Figure 6), which means more boronic groups are formed along the structure. The peaks around wavenumber 5199 cm^-1 belong to Al-O-H groups reduce depending on the molar ratio of boron used in the reaction (Figure-7), which shows successful formation of Al-O-B groups similar to Si-O-B groups. The peaks representing B-OH groups are found in the wavenumber region 4490-4089 cm ¹. The peaks are more intense in that region in case of DSZB and DAZB when compared to their corresponding DSZ and DAZ precursors (Figure 7). Even though they do not possess B-OH groups, the response of DAZ and DSZ in the region of 4089 - 4490 crrr ¹ is due to the interactions between silanol groups and water (Si-OH — H-O-H) molecules (Figure 7). In Figure 9, the responses of functional groups belongs to boron derivative zeolite between the wavenumbers 450 - 1600 cm ¹ can be seen.

In Figure 10 and Figure 11 , FT-IR spectrums of DAZB and DSZB formed from DAZ and DSZ precursors can be found. Mutual peaks were observed from the formed DAZB and DSZB. The adsorption peak between 3200 - 3250 crrr¹ wavenumber belongs to B-OH group. The adsorption peak between 500 - 900 cm^-1 wavenumber belongs to B- O-Si group.

In Figure 12, the FT-IR spectrums of the boron derivative zeolite materials formed by using different boric acid ratios can be seen. The issued peak intensities change in parallel to the amount of boric acid. The peaks representing B-0-X(FI, Si) and B-0 groups of DAZB formed from DAZ precursor under the optimized experimental conditions show higher intensities than the ones related to DSZB formed from equimolar DSZ precursor.

In Figure 13 and Figure 14, the SEM images of DAZB and DSZB which are produced under optimized conditions in 10pm and 30pm, , are represented.

In Figure 15, surface area (BET), particle size, boron content and pore volume of the adsorbents were investigated and results can be seen from the table.

Ill) Development of SPE cartridges by using DAZB and DSZB and the separation procedure of colorants and sugar

Boron derivative zeolites (DAZB and DSZB) are presented for the first time as chromatographic adsorbents to the separation process of the colorant mixture in the sugar and related products. The colorant compounds named as Sunset Yellow and Patent Blue V are separated from sucrose and both were isolated pure by using suitable eluents through the solid phase extraction (SPE) cartridges, which are filled with improved boron derivative zeolites as adsorbent. The successful purification of the sugar (sucrose) and colorants were achieved by boron derivatives zeolites due to the efficient combination of the ºSi-0-B(OH)_2, ºSi-OH, ºAI-0-B(OH)₂, XAI-OH chemical groups and porosity along their structure. The sugar compound sucrose is retained by its chemical interactions with the ºSi-0-B(OH)₂ groups of the DAZB or DSZB. On the other hand, the colorants (Sunset Yellow, Patent Blue V) are separated not only because of their different characteristic weak interactions between the unboronated free hydroxyl groups of ºSi-OH and XAI-OH on the backbone of the adsorbents but also the intense retention effect resulted by the high surface area porous skeleton of the DAZB and DSZB.

After the easy removal of the colorants in the mixture of the sugar-end product using the SPE cartridge filled with boron-derived zeolite using the appropriate pH mobile phase (pH>6), sugars retained in SPE are removed from boron-derived zeolite filler by adjusting the pH of the eluent to 5 (pH (5). Hence, the developed DAZB (or DSZB) type adsorbents posses chromatographic and affinity properties. Our studies lead to develop the novel DAZB or DSZB type adsorbents, and their SPE cartridges and HPLC columns.

Filling the SPE cartridges: SPE cartridges are developed by using 0.1 times by weight DAZB or DSZB adsorbent (filler) to 1 volume SPE cartridge. For this purpose, 0.1 g of boron-derived zeolite filler is filled into the unit volume of empty and fritted SPE cartridges with volume of 3, 6, 12 and 20 cc. (for example, 0.3g of absorbents is used for 3 volume of SPE cartridge, 0.6 of adsorbents for 6 volume of SPE cartridge). Pressure from top of the filled material is applied under vacuum in order to remove the empty space and fill the cartridges properly. With the help of a vacuum manifold, the SPE is conducted in four stages as below:

Conditioning: Water and methanol were applied 3-5 times (preferably 4 times) of the volume of the corresponding cartridge. At this stage, the elution speed of the mobile phase is also optimized and adjusted as 1 d/s (drop/second), 2 d/s, 3 d/s by tuning the vacuum.

Loading: The amount of the loading for the mixtures to be separated is 1 %, 2% and 5% of the adsorbent amount in the corresponding cartridges. The sugar sample to be separated is containing Patent Blue V, Sunset Yellow and Sucrose mixture. The sample is slowly loaded as its solution in water and elution speed is again optimized as 0.5d/s, 1 d/s, 2d/s.

Washing: Approximately 1/3 of the cartridge volume of methanol and hexane is passed through respectively. Elution: Elution solvent is added when the stopcock is closed. A clean test tube is placed in the manifold. 1 mL of 0.1 M ammonium acetate buffer solution (pH=7.5) is passed through the cartridge and collected in the test tube. The optimization studies for elution amount are carried out with 2 and 4 mL. Elution flow rate optimization studies are employed by 0.5 d/s, 1 d/s and 2 d/s. The collected eluate is analyzed by using HPLC for colorants and sugar. It is detected that the colorants have passed through the cartridge but sucrose is retained on the adsorbent when eluent has a pH>6. In following, sucrose could be detected by HPLC after changing the conditions of eluent to pH<5.

Recovery studies: A 12 cc fritted cartridge is filled with 1 g DAZB or DSZB. 20 mL methanol is passed through the cartridge under vacuum. In following, 25% w/w (0.05 g) colorant mixture (Sunset Yellow and Patent Blue V as content) immersed 0.2 g white sugar sample is solved in 0.2 mL of water and is applied to SPE cartridge . Then, methanol/acetonitrile (9: 1 ) mixture at pH>6 used as eluent to collect colorants as 1 mL fractions at 0.5 d/s flow rate. Afterwards, the same mobile phase is set to pH<5 and sucrose is obtained. All of the fractions are analyzed by HPLC-RID (sugar) and HPLC- DAD (colorants) for sugar and colorant content. The recovery of colorants and sugars obtained from SPEs are given in Table-2.

HPLC analysis showed that sugars are retained in SPE cartridge filled using DAZB or DSZB and dyes are recovered on average by 98%. The adsorbed sugar (sucrose) in the SPE cartridge is then recovered by setting the mobile phase to pH=5 by using sodium acetate buffer. The yield for the recovery of the sugar is calculated as 88% in the end of the process.

Considering these results, DAZB and DSZB are defined as an ideal adsorbant for separation of sugar and colorants mixtures.

As it was mentioned before, HPLC columns areprepared by using DAZB and DSZB adsorbents. The adsorbents are filled to the columns by using HPLC column filling machines. The self flushing solutions of 100% isopropyl alcohol (IPA), 100% methanol, 80/20 (v/v) IPA/water, and 80/20 (v/v) methanol/water are prepared before column filling proocess.

Hexane is passed before filling the column to avoid air bubbles in the pump. In following, the columns are filled with the synthesized DAZB and DSZB.

For this process, below steps were followed:

Weigh 3 g of the synthesized filler into a beaker and add 20 mL of hexane and mix.. Then,

• The mixture is kept in ultrasonic bath for 5 minutes.

• The mixture is quickly filled to the reservoir which was connected to the column.

• From the software of the machine, and a "pressure gradient" method is formed (flow rate and pressure optimization studies are carried out with hexane at 500 psi and 1000 psi for 10 minutes, followed by hexane at 2000 psi for 30 minutes. i).

• start the method

• The solvent coming from the column is sent to waste in this step.

• To make sure if the filling is properly made, 2D and 3D graphs are checked during solvent flaw by choosing "log data" tab. • In the end, hexane is passed through the column for 30 minutes in total. The column is removed from the machine, and the open ends were closed by caps to prevent the filling material to dry.

Industrial Use of the Invention

The invention can be used in the small scale R&D departments or industrial scale chromatographic purification processes of related sectors just as directly a chromatographic filling material of boron derivatives, the products such as HPLC columns (or any other liquid chromatography columns) of boron derivative zeolite materials. These products can also be designed custom-made columns of boron derivative zeolite columns materials in different sizes to meet different requirements in the industry can also be produced for the related sectors.

Claims

1. A chromatographic filler comprising desilicated or dealuminated ZSM-5, X or A type zeolite with boric acid, boric acid ethyl alcohol ester or boric acid methyl alcohol ester.

2. A method of production of DAZB (23.1 ) chromatographic filler comprising:

- dealumination of ZSM-5 type with high Si / Al ratio (Si / Al>50) or X or A type with low Si / Al ratio (Si / Al<5); and

- borylation of dealuminated zeolite with boric acid, boric acid ethyl alcohol esters, or boric acid methyl alcohols.

3. A method of production of DSZB (23.2) chromatographic filler comprising:

- ZSM-5 type with high Si / Al ratio (Si / Al>50) or X or A type with low Si / Al ratio (Si / Al<5); and

- borylation of desilicated zeolite with boric acid, boric acid ethyl alcohol esters, or boric acid methyl alcohols.

4. The method according to claim 2 comprising:

- adding 40-70 g of ZSM-5, X or A type zeolite (1 ) into reaction vessel,

- initiation of dealumination reaction (A) by adding 0.1 -8 N HCI, H2SO4, or HNO3 (2) 1 -4 times by volume of the zeolite,

- performing the dealumination reaction (A) at 70-180 °C (3.1 ) for 1 -24 hours under stirring and cooling (4) the mixture to room temperature to obtain acidic solution (5.1 ),

- performing filtration (B) by adding deionized water (6) 1 -4 times by volume of the zeolite to obtain the filtrate (7.1 ),

- 1 -3 times washing (C) the filtrate (7.1 ) by adding deionized water (6) 1 -4 times by volume of the filtrate,

- removing acid (8) and obtaining DAZ (9.1 ),

- drying DAZ (D) at 110-130 °C (10) during 12 hours,

- obtaining cooled DAZ (11.1 ) with cooling down (E) to room temperature (4) under vacuum (12),

- adding 0.5-1 moles of boric acid, boric acid ethyl alcohol esters and boric acid methyl alcohol esters (14) to 40-70 g DAZ (13.1 ) and 1 -3 times xylene (15) by volume of DAZ (13.1 ), - obtaining the mixture (17.1 ) which includes DAZ by borylation (F) of DAZ with boric acid, boric acid ethyl alcohol esters and boric acid methyl alcohol esters (14) with heating (16) at 170 - 200°C under vacuum (12),

- performing filtration (B) of the reaction mixture under vacuum (12) and remove xylene from the filtrate (18.1 ),

- washing (G) the filtrate by adding dichloromethane (19) 1 -3 times by volume of the filtrate (18.1 ) and obtaining the filtrate (20.1 ); and

- drying the filtrate (20.1 ) at 80-120 °C (21 ) for 12 hours (H) and cooling (I) the dried mixture (22.1 ) to room temperature (4) in a vacuum (12).

5. The method according to claim 3 comprising:

- adding 40-70 g of ZSM-5, X or A type zeolite (1 ) into reaction vessel,

- initiation of desilication reaction (J) by adding 0.1 -4 N NaOH or KOH (24) 1 -4 times by volume of the zeolite,

- performing the desilication reaction (J) at 100-130 °C (3.2) for 3-12 hours (3.1 ) under stirring and cooling (4) the mixture to room temperature to obtain alkaline solution (5.2),

- performing filtration (B) by adding deionized water (6) 1 -4 times by volume of the zeolite to obtain filtrate (7.2),

- 1 -3 times washing (C) the filtrate (7.2) by adding deionized water (6) 1 -4 times by volume of the filtrate,

- removing alkaline solution (25) and obtaining DSZ (9.2),

- drying DSZ (D) at 110-130 °C (10) during 12 hours,

- obtaining dried DSZ (13.2) by cooling down (E) to room temperature (4) under vacuum (12),

- adding 0.5-1 moles of boric acid, boric acid ethyl alcohol esters and boric acid methyl alcohol esters (14) to 40-70 g DSZ (13.2) and 1 -3 times xylene (15) by volume of DSZ (13.2),

- obtaining the mixture (17.2) which includes DSZ by borylation (F) of DAZ with boric acid, boric acid ethyl alcohol esters and boric acid methyl alcohol esters (14) with heating (16) at 170 - 200°C under vacuum (12),

- performing filtration (B) of the reaction mixture under vacuum (12) and remove xylene from the filtrate (18.2),

- washing (G) the filtrate by adding dichloromethane (19) 1 -3 times by volume of the filtrate (18.2) and obtaining the filtrate (20.2); and - drying the filtrate (20.2) at 80-120 °C (21 ) for 12 hours (H) and cooling (I) the dried mixture (22.2) to room temperature (4) in a vacuum (12).

6. A chromatographic extraction (SPE) cartridge comprising DAZB (23.1 ) or DSZB (23.2) as filler.

7. A high-pressure liquid chromatography (HPLC) column comprising DAZB (23.1 ) or DSZB (23.2) as filler.

8. A method for the separation of the mixture of colorant-sugar comprising:

- filling 3-20 cc volume SPE cartridge using 0.1 g DAZB (23.1 ) or DSZB (23.2) per unit volume of SPE cartridge,

- conditioning of the cartridge with 3-5 volume of water and methanol by passing 1 -3 d/s elution speed,

- Loading the 1 -5% of DSZB or DAZB of sugar-colorant mixture into the cartridge at a flow rate of 0.5-2 d/s,

- Washing respectively by passing 1/3 volume of the cartridge of methanol and hexane; and

- eluating by adding 1 -4 mL of 0.1 M acetate buffer at a flow rate of 0.5-2 d/s.