Home Study on the preparation of glycerylphosphorylcholine by transesterification under supported sodium methoxide
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Study on the preparation of glycerylphosphorylcholine by transesterification under supported sodium methoxide

  • Yiwen Rao , Lele Zhou , Zejing Fan , Hongya Li EMAIL logo , Biao Yan and Xiaoli Zhang
Published/Copyright: December 9, 2023
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Abstract

Glycerylphosphorylcholine (GPC) was prepared by transesterification using supported sodium methoxide as catalyst and natural lecithin as raw material. Sodium methoxide has been proved to be an effective catalyst for the preparation of GPC, which is easy to recover and reuse. After six repeated uses, its stability is satisfactory. The effects of agitation speed, catalyst dosage, and reaction temperature on the reaction were investigated, respectively, and the optimum conditions for preparing GPC catalyzed by supported sodium methoxide were found: the concentration of phosphorylcholine was 0.1 mol·L−1, the stirring speed was 600 rpm, the amount of catalyst was 7.5 g·L−1, the reaction temperature was 45°C, and the reaction time was 4 h; then, the conversion rate of phosphatidylcholine could reach 99%. At the same time, the reaction kinetic model was established based on the mechanism of the transesterification, and the experimental data were compared with the calculated values; it was found that the experimental data fitted the model well. Finally, the reaction activation energy obtained by the Arrhenius equation is 41.6 kJ·mol−1, which indicates that the supported sodium methoxide has good catalytic performance in this reaction system.

1 Introduction

Glycerylphosphorylcholine (GPC) is a water-soluble phospholipid precursor that is found in the human body, and it is very important for treating brain diseases such as acute cerebrovascular accident, Alzheimer’s disease, multi-infarct dementia, and cerebral circulation decline, and it can also improve the attention, cognition, and memory of all ages [1,2,3]. In addition, GPC has the largest effects on protecting the liver and kidney, maintaining the normal secretion of reproductive hormones, and reducing high blood lipid. So, it is widely used in food, health products, medicine, and other industries, which has greatly improved people’s quality of life [4,5,6,7]. However, the content of GPC in nature is relatively small, and the domestic research on GPC started late and developed slowly; there is a certain difference in quality scale with foreign products. Therefore, many scholars and companies have invested in the research of preparing and producing high-purity GPC, with the hope that it will bring great medicinal value and economic value [8,9].

There are several methods for preparing GPC, the biological extraction method [10,11], in which the raw material is limited, the process is complicated, and the cost is high; in the chemical synthesis method [12,13], which has many side reactions, the intermediate products are unstable and the product yield is lower, while the transesterification method has attracted widespread attention of scholars at home and abroad due to its natural and reliable raw materials in recent years [14,15,16,17,18,19,20]. Chadha [21] used egg yolk lecithin as raw material, added cadmium chloride in the alcoholysis process, and finally obtained a complex of GPC and cadmium chloride through recrystallization, in order to obtain pure product; it still needed further treatment. Kangyi et al. [22] hydrolyzed soybean lecithin powder using phospholipase as a catalyst to obtain GPC, and the product was separated using a chromatography column. The purity and recovery of the final GPC were 99.8% and 69.8%, respectively. Our research group has carried out a series of studies on GPC preparation under different catalysts [23,24,25], and the properties of several catalysts are shown in Table 1. In the preparation of GPC, the catalyst used affects the difficulty of separation and purity of product directly, and the solid catalyst can be recycled many times and has long service life [26,27,28], so the choice of catalyst and its preparation process are very important for the preparation of GPC by transesterification.

Table 1

Comparison of the performance of several catalysts

Literature number Catalyst used Reaction conditions Conversion of PC Advantage and disadvantage
[23] Low-boiling point organic amines Catalyst content: 90% Advantage:
2.4% of the total reaction volume (1) Good activity
Lecithin concentration: 0.10 mol·L−1 (2) Easy to separate
Reaction temperature: 60°C (3) Little deactivation
Reaction time: 4 h Disadvantage:
Low conversion rate
[24] Propylamine Catalyst content: 98% Advantage:
2.4% of the total reaction volume (1) Easy to separate
Lecithin concentration: (2) Easy to recycle
0.05–0.10mol·L−1 Disadvantage: Short lifespan
Reaction temperature: 60°C
Reaction time: 4h
[25] Tert-butylamine Catalyst content: 98% Advantage:
2.4% of the total reaction volume (1) High purity
Lecithin concentration: 0.10mol·L−1 (2) Little deactivation
Reaction temperature: 60°C (3) Long service life
Reaction time: 4h Disadvantage:
Not easy to recycle
[15] Calcined sodium silicate Catalyst content: 6 wt% 99.50% Advantage:
Lecithin concentration: (1) Easy to separate
10.5mmol·L−1 (2) Repeated use
Reaction temperature: 65°C Still maintain high activity
Reaction time: 2h (3) High conversion rate

In this work, GPC is prepared by transesterification using lecithin as raw material, catalyzed by the supported sodium methoxide. The optimal conditions for the reaction were obtained, and the reaction kinetics was discussed in order to provide theoretical basis for its practical application.

2 Experimental sections

2.1 Materials and chemicals

Lecithin was purchased from Tianjin Heowns Biochemical Technology Co., Ltd. All the chemicals such as sodium methoxide, methanol, and chloroform are of analytical grade, and water used is deionized water.

2.2 Preparation of catalyst

A certain amount of activated silica was added to a round-bottomed flask, and the sodium methoxide solution (2.2 mol·L−1) in methanol was poured into it at the ratio of 0.25 g·mL−1, then stirred, and refluxed at 60°C for 4 h, and filtered after cooling. Finally, the supported sodium methoxide catalyst was obtained after dried.

2.3 Transesterification reaction

The transesterification was performed in a 100 mL three-necked flask equipped with a reflux condenser and magnetic stirrer. When the methanol solution of lecithin was heated to the setting temperature, the supported sodium methoxide catalyst was added and the stirrer was started, and liquid samples about 0.5 mL were withdrawn by a syringe in predefined time intervals; the sample solutions were collected in small vials for further analysis.

2.4 Analysis method

Thin-layer chromatography (TLC) was used to analyze the sampling and calculate the conversion of phosphatidylcholine (PC). Each sample with equal volume was spotted on the TLC plate, which was prepared from our laboratory by silica gel GF254. After drying for several minutes, the plate was developed in the solvent of chloroform:methanol:water = 13:5:0.8 (volume ratio), the developed plate was dried in an oven for about 10 min and detected in the iodine chamber, and then yellow spots with white background appeared after several minutes. The spots were quantified by densitometric scanning at 450 nm using a Shimadzu dual-wavelength chromato scanner CS-930.

3 Results and discussion

3.1 Effect of stirring speed

For heterogeneous reaction processes, the stirring speed affects the mixing, diffusing, and contacting with catalyst of the reactants. Increasing the speed of the stirrer can reduce the effect of external diffusion on the reaction. The concentration of PC is 0.1 mol·L−1, the catalyst dosage is 10 g·L−1, the reaction temperature is 40°C, and the reaction time is 4 h; the effect of stirring speed on the transesterification was examined at 200, 400, 600, and 800 rpm, respectively, and the results are shown in Figure 1.

Figure 1 
                  Effect of stirring speed on the conversion of PC. Reaction conditions: the concentration of PC was 0.1 mol·L−1, the catalyst dosage was 10 g·L−1, the reaction temperature was 40°C, and the reaction time was 4 h.
Figure 1

Effect of stirring speed on the conversion of PC. Reaction conditions: the concentration of PC was 0.1 mol·L−1, the catalyst dosage was 10 g·L−1, the reaction temperature was 40°C, and the reaction time was 4 h.

It was found that with increasing stirring speed, the conversion of PC increased, but when the stirring speed was greater than 600 rpm, the stirring speed had little effect on the conversion of PC, indicating that the catalyst was distributed satisfactory in the reactor at the high stirring speed, and this was considered to be the minimum stirring speed required to eliminate the effect of external diffusion, so the 600 rpm was selected at the subsequent experiments.

3.2 Effect of catalyst amount

As an important factor affecting the chemical reaction, the catalyst not only affects the speed of the reaction and the yield of the product, but also affects the difficulty of purification and separation of subsequent products seriously. Under the conditions of PC concentration of 0.1 mol·L−1, stirring speed of 600 rpm, reaction temperature of 40°C, and reaction time of 4 h, the catalyst amount was investigated at 5, 7.5, 10, and 12.5 g·L−1, and the results are shown in Figure 2.

Figure 2 
                  Effect of catalyst amount on the conversion of PC. Reaction conditions: concentration of PC was 0.1 mol·L−1, stirring speed was 600 rpm, reaction temperature was 40°C, and reaction time was 4 h.
Figure 2

Effect of catalyst amount on the conversion of PC. Reaction conditions: concentration of PC was 0.1 mol·L−1, stirring speed was 600 rpm, reaction temperature was 40°C, and reaction time was 4 h.

It can be seen that the conversion of PC increased with the increase in catalyst amount. When the amount of catalyst exceeded 7.5 g·L−1, the amount of catalyst had no significant effect on the conversion of PC. At the end of the reaction, the conversion of PC under catalyst amount of 12.5 g·L−1 was only 4% higher than that at 7.5 g·L−1. Therefore, the appropriate amount of catalyst used in the subsequent experiments was 7.5 g·L−1.

3.3 Effect of reaction temperature

The reaction temperature was examined at 30°C, 35°C, 40°C, 45°C, and 50°C, respectively, to discuss its effect on the transesterification process, under the conditions of a concentration of PC of 0.1 mol·L−1, a stirring speed of 600 rpm, a catalyst amount of 7.5 g·L−1, and a reaction time of 4 h. The results are shown in Figure 3.

Figure 3 
                  Effect of temperature on the conversion of PC. Reaction conditions: concentration of PC was 0.1 mol·L−1, stirring speed was 600 rpm, catalyst amount was 7.5 g·L−1, and reaction time was 4 h.
Figure 3

Effect of temperature on the conversion of PC. Reaction conditions: concentration of PC was 0.1 mol·L−1, stirring speed was 600 rpm, catalyst amount was 7.5 g·L−1, and reaction time was 4 h.

As can be seen, the conversion of PC increased with increasing temperature. When the temperature rose above 40°C, the increase in the conversion of PC gradually decreased. When the reaction temperatures were 45°C and 50°C, the conversion of PC was basically the same at the end of the reaction. Therefore, 45°C was selected as the most suitable experimental temperature.

3.4 Stability of catalyst

When the transesterification completed under the suitable conditions, the catalyst was filtered off and reused after washing with anhydrous methanol (Figure 4). It was found that only 9% decrease in PC conversion was observed after recycling six times under the same conditions, and the catalyst stability was satisfactory during the experiment.

Figure 4 
                  Stability of the catalyst.
Figure 4

Stability of the catalyst.

3.5 Reaction kinetic analysis

The mechanism of the transesterification can be interpreted as shown in Scheme 1. First, sodium methylate dissociates the methoxy ion. In the second step, the nucleophilic methoxy group attacks the carbonyl carbon atom on the PC molecule and forms an intermediate with a tetrahedral structure. In the third step, this intermediate undergoes electron transfer and rearranges to produce lysophospholipid (LPC) negative ions, which combine with protons provided by methanol to form the intermediate LPC. Finally, the methoxy group continues to attack the acyl carbon atoms on the lecithin molecule, and the process is repeated to produce GPC.

Scheme 1 
                  Mechanism of transesterification catalyzed by the supported sodium methoxide.
Scheme 1

Mechanism of transesterification catalyzed by the supported sodium methoxide.

The entire transesterification can be expressed as follows:

(1) PC + MeOH k 1 k 1 LPC + Ester

(2) LPC + MeOH k 2 k 2 GPC + Ester

The first step is assumed to be much faster than the second step, so the intermediate LPC is in equilibrium with the reactants PC and MeOH at any time. Using the steady-state approximation, the concentration of LPC can be expressed as:

(3) d C L d t = k 1 C P C M k 1 ' C L C E = 0

so,

(4) C L = k 1 C P C M k 1 C E

During the entire reaction process, methanol is greatly excessive, so the reverse reaction can be ignored. The formation rate of GPC can be expressed as:

(5) d C G d t = k 2 C L C M

Expressing the concentration of each substance by the conversion rate of PC, x p, so the formula 5 can be changed to:

(6) d x P d t = k 1 k 2 ( 1 x P ) ( C M 0 2 C P 0 x P ) 2 k 1 C P 0 2 x P

Defined C M 0 / C P 0 = M, K = k 1 k 2 / k 1 , then

(7) d x P d t = K C P 0 ( 1 x P ) ( M 2 x P ) 2 2 x P

By integrating the aforementioned formula, we get

(8) K C P0 t = K t = 2 M 2 1 M 2 ln M 2 x P 1 x P M 2 ( M 2 x P )

The experimental data under different catalyst amounts were substituted into the right part of formula 8 and plotted against the reaction time. It was found that the data fit the equation model well (Figure 5).

Figure 5 
                  Fitting of reaction kinetics at different catalyst amounts.
Figure 5

Fitting of reaction kinetics at different catalyst amounts.

Similarly, the experimental data at different reaction temperatures were substituted into the right part of formula 8 and plotted against the reaction time (Figure 6). The linear correlation coefficient R 2 of each fitted line is above 0.97, which indicates that the kinetic equation model is equally applicable for the reaction systems at different temperatures.

Figure 6 
                  Fitting of reaction kinetics at different reaction temperatures.
Figure 6

Fitting of reaction kinetics at different reaction temperatures.

According to the Arrhenius equation, the logarithms of the slope k for the dynamic fit curves at different temperatures were plotted against 1/T (Figure 7). From the slope of the straight line, the activation energy E a is 41.6 kJ·mol−1, indicating that the catalyst has good catalytic performance in their action system.

Figure 7 
                  Arrhenius plot of ln K vs 103/T.
Figure 7

Arrhenius plot of ln K vs 103/T.

4 Conclusions

The supported sodium methoxide catalyst has achieved significant results in the preparation of GPC by transesterification. Under the optimum reaction conditions, the amount of catalyst is 7.5 g·L−1, the reaction temperature is 45°C, stirring rate is 600 rpm, and reaction time is 240 min, and the conversion rate of PC is 98.9%. The reaction kinetic model was established, and it was found that the experimental data fitted the model well. The activation energy of the reaction was 41.6 kJ·mol−1, and the catalytic performance of the supported sodium methoxide in this reaction system is satisfactory.

  1. Funding information: This work was funded by the National Natural Science Foundation of China (Award No. 22163012) and the Key Laboratory Project Foundation of Shaanxi Provincial Education Department in China (Award No. 20JS158).

  2. Author contributions: Yiwen Rao: writing – original draft, validation, methodology, formal analysis, visualization; Hongya Li: writing – review and editing, funding acquisition, resources, methodology, supervision, project administration; Lele Zhou: resources, data curation, investigation; Zejing Fan: formal analysis, investigation, resources; Biao Yan: validation, formal analysis, data curation; Xiaoli Zhang: visualization, investigation, formal analysis.

  3. Conflict of interest: The authors state no conflict of interest.

  4. Data availability statement: All data generated or analysed during this study are included in this published article.

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Received: 2023-06-26
Accepted: 2023-10-01
Published Online: 2023-12-09

© 2023 the author(s), published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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  63. Biosynthesis of zinc oxide nanoparticles from molted feathers of Pavo cristatus and their antibiofilm and anticancer activities
  64. Clean preparation of rutile from Ti-containing mixed molten slag by CO2 oxidation
  65. Synthesis and characterization of Pluronic F-127-coated titanium dioxide nanoparticles synthesized from extracts of Atractylodes macrocephala leaf for antioxidant, antimicrobial, and anticancer properties
  66. Effect of pretreatment with alkali on the anaerobic digestion characteristics of kitchen waste and analysis of microbial diversity
  67. Ameliorated antimicrobial, antioxidant, and anticancer properties by Plectranthus vettiveroides root extract-mediated green synthesis of chitosan nanoparticles
  68. Microwave-accelerated pretreatment technique in green extraction of oil and bioactive compounds from camelina seeds: Effectiveness and characterization
  69. Studies on the extraction performance of phorate by aptamer-functionalized magnetic nanoparticles in plasma samples
  70. Investigation of structural properties and antibacterial activity of AgO nanoparticle extract from Solanum nigrum/Mentha leaf extracts by green synthesis method
  71. Green fabrication of chitosan from marine crustaceans and mushroom waste: Toward sustainable resource utilization
  72. Synthesis, characterization, and evaluation of nanoparticles of clodinofop propargyl and fenoxaprop-P-ethyl on weed control, growth, and yield of wheat (Triticum aestivum L.)
  73. The enhanced adsorption properties of phosphorus from aqueous solutions using lanthanum modified synthetic zeolites
  74. Separation of graphene oxides of different sizes by multi-layer dialysis and anti-friction and lubrication performance
  75. Visible-light-assisted base-catalyzed, one-pot synthesis of highly functionalized cinnolines
  76. The experimental study on the air oxidation of 5-hydroxymethylfurfural to 2,5-furandicarboxylic acid with Co–Mn–Br system
  77. Highly efficient removal of tetracycline and methyl violet 2B from aqueous solution using the bimetallic FeZn-ZIFs catalyst
  78. A thermo-tolerant cellulase enzyme produced by Bacillus amyloliquefaciens M7, an insight into synthesis, optimization, characterization, and bio-polishing activity
  79. Exploration of ketone derivatives of succinimide for their antidiabetic potential: In vitro and in vivo approaches
  80. Ultrasound-assisted green synthesis and in silico study of 6-(4-(butylamino)-6-(diethylamino)-1,3,5-triazin-2-yl)oxypyridazine derivatives
  81. A study of the anticancer potential of Pluronic F-127 encapsulated Fe2O3 nanoparticles derived from Berberis vulgaris extract
  82. Biogenic synthesis of silver nanoparticles using Consolida orientalis flowers: Identification, catalytic degradation, and biological effect
  83. Initial assessment of the presence of plastic waste in some coastal mangrove forests in Vietnam
  84. Adsorption synergy electrocatalytic degradation of phenol by active oxygen-containing species generated in Co-coal based cathode and graphite anode
  85. Antibacterial, antifungal, antioxidant, and cytotoxicity activities of the aqueous extract of Syzygium aromaticum-mediated synthesized novel silver nanoparticles
  86. Synthesis of a silica matrix with ZnO nanoparticles for the fabrication of a recyclable photodegradation system to eliminate methylene blue dye
  87. Natural polymer fillers instead of dye and pigments: Pumice and scoria in PDMS fluid and elastomer composites
  88. Study on the preparation of glycerylphosphorylcholine by transesterification under supported sodium methoxide
  89. Wireless network handheld terminal-based green ecological sustainable design evaluation system: Improved data communication and reduced packet loss rate
  90. The optimization of hydrogel strength from cassava starch using oxidized sucrose as a crosslinking agent
  91. Green synthesis of silver nanoparticles using Saccharum officinarum leaf extract for antiviral paint
  92. Study on the reliability of nano-silver-coated tin solder joints for flip chips
  93. Environmentally sustainable analytical quality by design aided RP-HPLC method for the estimation of brilliant blue in commercial food samples employing a green-ultrasound-assisted extraction technique
  94. Anticancer and antimicrobial potential of zinc/sodium alginate/polyethylene glycol/d-pinitol nanocomposites against osteosarcoma MG-63 cells
  95. Nanoporous carbon@CoFe2O4 nanocomposite as a green absorbent for the adsorptive removal of Hg(ii) from aqueous solutions
  96. Characterization of silver sulfide nanoparticles from actinobacterial strain (M10A62) and its toxicity against lepidopteran and dipterans insect species
  97. Phyto-fabrication and characterization of silver nanoparticles using Withania somnifera: Investigating antioxidant potential
  98. Effect of e-waste nanofillers on the mechanical, thermal, and wear properties of epoxy-blend sisal woven fiber-reinforced composites
  99. Magnesium nanohydroxide (2D brucite) as a host matrix for thymol and carvacrol: Synthesis, characterization, and inhibition of foodborne pathogens
  100. Synergistic inhibitive effect of a hybrid zinc oxide-benzalkonium chloride composite on the corrosion of carbon steel in a sulfuric acidic solution
  101. Review Articles
  102. Role and the importance of green approach in biosynthesis of nanopropolis and effectiveness of propolis in the treatment of COVID-19 pandemic
  103. Gum tragacanth-mediated synthesis of metal nanoparticles, characterization, and their applications as a bactericide, catalyst, antioxidant, and peroxidase mimic
  104. Green-processed nano-biocomposite (ZnO–TiO2): Potential candidates for biomedical applications
  105. Reaction mechanisms in microwave-assisted lignin depolymerisation in hydrogen-donating solvents
  106. Recent progress on non-noble metal catalysts for the deoxydehydration of biomass-derived oxygenates
  107. Rapid Communication
  108. Phosphorus removal by iron–carbon microelectrolysis: A new way to achieve phosphorus recovery
  109. Special Issue: Biomolecules-derived synthesis of nanomaterials for environmental and biological applications (Guest Editors: Arpita Roy and Fernanda Maria Policarpo Tonelli)
  110. Biomolecules-derived synthesis of nanomaterials for environmental and biological applications
  111. Nano-encapsulated tanshinone IIA in PLGA-PEG-COOH inhibits apoptosis and inflammation in cerebral ischemia/reperfusion injury
  112. Green fabrication of silver nanoparticles using Melia azedarach ripened fruit extract, their characterization, and biological properties
  113. Green-synthesized nanoparticles and their therapeutic applications: A review
  114. Antioxidant, antibacterial, and cytotoxicity potential of synthesized silver nanoparticles from the Cassia alata leaf aqueous extract
  115. Green synthesis of silver nanoparticles using Callisia fragrans leaf extract and its anticancer activity against MCF-7, HepG2, KB, LU-1, and MKN-7 cell lines
  116. Algae-based green AgNPs, AuNPs, and FeNPs as potential nanoremediators
  117. Green synthesis of Kickxia elatine-induced silver nanoparticles and their role as anti-acetylcholinesterase in the treatment of Alzheimer’s disease
  118. Phytocrystallization of silver nanoparticles using Cassia alata flower extract for effective control of fungal skin pathogens
  119. Antibacterial wound dressing with hydrogel from chitosan and polyvinyl alcohol from the red cabbage extract loaded with silver nanoparticles
  120. Leveraging of mycogenic copper oxide nanostructures for disease management of Alternaria blight of Brassica juncea
  121. Nanoscale molecular reactions in microbiological medicines in modern medical applications
  122. Synthesis and characterization of ZnO/β-cyclodextrin/nicotinic acid nanocomposite and its biological and environmental application
  123. Green synthesis of silver nanoparticles via Taxus wallichiana Zucc. plant-derived Taxol: Novel utilization as anticancer, antioxidation, anti-inflammation, and antiurolithic potential
  124. Recyclability and catalytic characteristics of copper oxide nanoparticles derived from bougainvillea plant flower extract for biomedical application
  125. Phytofabrication, characterization, and evaluation of novel bioinspired selenium–iron (Se–Fe) nanocomposites using Allium sativum extract for bio-potential applications
  126. Erratum
  127. Erratum to “Synthesis, characterization, and evaluation of nanoparticles of clodinofop propargyl and fenoxaprop-P-ethyl on weed control, growth, and yield of wheat (Triticum aestivum L.)”
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