High Performance Liquid Chromatography-evaporative Light Research Articles

Increased Accumulation of Ginsenosides in Panax ginseng Sprouts Cultivated with Kelp Fermentates

Currently, new agri-tech has been developed and adapted for the cultivation of crops using smart farming technologies, e.g., plant factories and hydroponics. Kelp (Laminaria japonica), which has a high industrial value, was considered as an alternative to chemicals for its eco-friendly and sustainably wide use in crop cultivation. In this study, a fermented kelp (FK) was developed for use in hydroponics. The FK contained various free and protein-bound amino acid compositions produced by fermenting the kelp with Saccharomyces cerevisiae. Supplementing FK as an aeroponic medium when cultivating ginseng sprouts (GSs) elevated the total phenolic and flavonoid contents. Additionally, seven ginsenosides (Rg1, Re, Rb1, Rc, Rg2, Rb2, and Rd) in GSs cultivated with FK in a smart-farm system were identified and quantified by a high-performance liquid chromatography-evaporative light scattering detector/mass spectrometry analysis. Administering FK significantly increased the ginsenosides in the GSs compared to the control group, which was cultivated with tap water. These results indicate the FK administration contributed to the increased accumulation of ginsenosides in the GSs. Overall, this study suggests that FK, which contains abundant nutrients for plant growth, can be used as a novel nutrient solution to enhance the ginsenoside content in GSs during hydroponic cultivation.

A novel and sensitive method for simultaneous determination of 6 low-calorie bulk sweeteners by HPLC-ELSD

A novel and sensitive method for the simultaneous analysis of six low-calorie bulk sweeteners (D-allulose, D-tagatose, D-mannitol, mycose, palatinose, and erythritol) without derivatisation was developed using high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD). Chromatographic separations were carried out on a Zorbax Original NH2 (5 μm particle size, 250 mm×4.60 mm id, 70 Å) column with flow rate gradient elution with acetonitrile: water (80:20, v/v). Drift tube temperature was set at 50 ℃, the nebuliser carrier gas flow rate was 1.0 mL·min-1, and nitrogen pressure was regulated to 276 kPa with gain:3. The regression equation showed good linearity (R2 = 0.9985-0.9998) for all six low-calorie bulk sweeteners in the tested range (0.060-0.60 mg·mL-1). The limits of detection (LOD) for the six low-calorie bulk sweeteners ranged from 0.02 to 0.06 mg·mL-1. The proposed HPLC-ELSD method was validated for the quantification of the low-calorie bulk sweeteners in 14 types of foods, and the results were satisfactory. In addition, the results showed that the number of sweeteners in each food product varied. The presence of multiple low-calorie bulk sweeteners in certain foods is interesting. This method is successful in monitoring low-calorie bulk sweeteners in food.

The improvement of kinsenoside in wild-imitated cultivation Anoectochilus roxburghii associated with endophytic community

Due to the low content of active ingredients and poor efficacy in herbal medicine in modern large-scale planting conditions, a growing number of methods for improving the content of active ingredients have been studied, such as improving planting techniques, developing and applying microbial engineering bacteria. In recent years, most studies have focused on improving the quality of Anoectochilus roxburghii through conventional farming methods. However, the role of endophytes in improving the quality of herbal medicine was often ignored. Although some studies have shown that microorganisms can regulate Anoectochilus roxburghii growth, there are still few studies on their regulation of plant active ingredients in plants. In this study, endophytic community and active ingredient content under the three culture patterns were examined through amplicon sequencing and high performance liquid chromatography-evaporative light scattering detector, and the correlation between the abundance of endophytes and kinsenoside content was analyzed. The results indicated a significant alteration in the diversity and network complexity of endophytic community within Anoectochilus roxburghii under the three culture conditions. After being cultivated in a wild-imitated environment for 3 months, the microbial composition of wild-imitated cultivated Anoectochilus roxburghii differentiated and converged toward that of wild-tending planted Anoectochilus roxburghii. Correlation analyses showed that the diversity of bacterial community has a significant positive relationship with the content of kinsenoside, and Burkholderia-Caballeronia-Paraburkholderia is the key taxa affecting kinsenoside content. In addition, acetylacetyl-CoA reductase, a key enzyme for the synthesis of kinsenoside, was significantly enriched in wild-imitated cultivated and wild tending cultivated Anoectochilus roxburghii. The study proposed that the medicinal quality of wild-imitated cultivated and wild tending cultivated Anoectochilus roxburghii, with high content of kinsenoside, which is likely attributed to the Burkholderia-Caballeronia-Paraburkholderia and acetylacetyl-CoA. This study provides a basis for the improvement and sustainable utilization of Anoectochilus roxburghii resources.

Carbohydrate-based quality consistency evaluation of Saposhnikoviae Radix

This study aims to evaluate the quality consistency of Saposhnikoviae Radix based on carbohydrates, and explore the potential of carbohydrates as the internal quality control indicators of Saposhnikoviae Radix. The total polysaccharides were quantified by UV-Vis spectrophotometry and the molecular weight range of the polysaccharides was determined by high performance gel-permeation chromatography-evaporative light scattering detection(HPGPC-ELSD). The monosaccharides in polysaccharides and the free monosaccharides were quantified by high performance liquid chromatography-UV detection(HPLC-UV), and the oligosaccharides and fructose were quantified by high performance liquid chromatography-evaporative light scattering detection(HPLC-ELSD). The carbohydrate-based quality of Saposhnikoviae Radix was compared among 45 batches of commercial samples and 13 batches of self-collected samples. The results showed that the molecular weight distribution, monosaccharide composition, oligosaccharide, and free monosaccharide composition were similar in the 58 batches of samples. The average content of total polysaccharides, oligosaccharides, and total free monosaccharides in commercial samples were 39.66, 148.79, and 68.62 mg·g~(-1), respectively. The content showed significant differences among batches, with the highest differences of 3.51, 1.75, and 2.58 times, respectively. The RSD of the relative ratios of monosaccharides in the polysaccharides in commercial samples reached 28%-45%. The average content of total polysaccharides, oligosaccharides, and total free monosaccharides in self-collected samples were 68.07, 145.76, and 42.04 mg·g~(-1), respectively, with the inter-region differences of 2.88, 1.88, and 1.07 times, respectively. The RSD of the relative ratios of monosaccharides in polysaccharides in self-collected samples ranged from 8.2% to 59%. The total polysaccharides and total free monosaccharides in self-collected samples were 1.72 times higher and 1.63 times lower, respectively, than those in commercial samples. The content of oligosaccharides was similar between self-collected samples and commercial samples. To sum up, carbohydrates are one of the material bases for the internal quality consistency of Saposhnikoviae Radix. The qualitative characteristics of polysaccharides and the quantitative characteristics of polysaccharides and oligosaccharides are related to the origin of medicinal materials. Moreover, the quantitative characteristics of polysaccharides and free monosaccharides may be related to the storage conditions. Carbohydrates are potential indicators for the quality control of Saposhnikoviae Radix and deserve attention.

Developing advanced models of biological membranes with hydrogenous and deuterated natural glycerophospholipid mixtures

Cellular membranes are complex systems that consist of hundreds of different lipid species. Their investigation often relies on simple bilayer models including few synthetic lipid species. Glycerophospholipids (GPLs) extracted from cells are a valuable resource to produce advanced models of biological membranes. Here, we present the optimisation of a method previously reported by our team for the extraction and purification of various GPL mixtures from Pichia pastoris. The implementation of an additional purification step by High Performance Liquid Chromatography-Evaporative Light Scattering Detector (HPLC-ELSD) enabled for a better separation of the GPL mixtures from the neutral lipid fraction that includes sterols, and also allowed for the GPLs to be purified according to their different polar headgroups. Pure GPL mixtures at significantly high yields were produced through this approach. For this study, we utilised phoshatidylcholine (PC), phosphatidylserine (PS) and phosphatidylglycerol (PG) mixtures. These exhibit a single composition of the polar head, i.e., PC, PS or PG, but contain several molecular species consisting of acyl chains of varying length and unsaturation, which were determined by Gas Chromatography (GC). The lipid mixtures were produced both in their hydrogenous (H) and deuterated (D) versions and were used to form lipid bilayers both on solid substrates and as vesicles in solution. The supported lipid bilayers were characterised by quartz crystal microbalance with dissipation monitoring (QCM-D) and neutron reflectometry (NR), whereas the vesicles by small angle X-ray (SAXS) and neutron scattering (SANS). Our results show that despite differences in the acyl chain composition, the hydrogenous and deuterated extracts produced bilayers with very comparable structures, which makes them valuable to design experiments involving selective deuteration with techniques such as NMR, neutron scattering or infrared spectroscopy.

Determination of sugars and sugar alcohols in infant formula by high performance liquid chromatography with evaporative light-scattering detector

A method was established for the simultaneous determination of five sugars (fructose, glucose, sucrose, lactose, maltose) and five sugar alcohols (erythritol, xylitol, sorbitol, mannitol, maltitol) in infant formula by high performance liquid chromatography-evaporative light scattering detector. After the samples were extracted with acetonitrile–water solution, precipitated by acetic acid, and purified with solid phase extraction cartridge, ALLChrom Rocksil Carbohydrate ES column was adopted for separation, and isocratic elution was conducted at the flow rate of 1.0 mL/min with acetonitrile-0.04 % ammonia solution as the mobile phase. The analytes were detected by an evaporative light-scattering detector, and quantified by external standard method. The linear ranges of the 10 components were 0.04–4.0 g/L with the correlation coefficients greater than 0.999, and the limits of quantification (S/N = 10) of the method were 0.08–0.4 g/100 g. The relative standard deviation of the lactose parallel samples reached 1.29 %, and the recoveries of the other 9 components ranged from 80.4 % to 99.4 % with the relative standard deviation of 2.8 %-7.1 %. The method performs well in sensitivity and separation, which is suitable for the simultaneous quantitative determination of sugars and sugar alcohols in infant formula.

In vitro anti-obesity activity by pancreatic lipase inhibition - Simple HPLC approach using EVOO as natural substrate.

Pancreatic lipase (PL) is a key lipolytic enzyme in humans for the digestion and absorption of dietary fats. Thereby, PL is a well-recognized target in the management of obesity and its inhibition attracts the interest of researchers globally. The screening of new natural PL inhibitors as alternative strategy to the synthesis of chemical ones represents nowadays a hot topic in research. The main challenge in this matter is the lack of a universal analytical method allowing the monitoring of PL activity and the reliable quantification of lipid digestion products. The (normal phase)-high-performance liquid chromatography-evaporative light scattering detector [(NP)-HPLC-ELSD] method proposed in this work represents a direct and rapid strategy to simultaneously quantify the products obtained from in vitro PL digestion. As one of the main novelties, the triacylglycerol (TAG) fraction from extra-virgin olive oil was selected as natural substrate. The PL activity was measured by monitoring the levels of remaining TAGs and formed free fatty acids (FFAs), using Orlistat as known inhibitor. The method validation confirmed the adequacy of the analytical method for quantitative purposes, showing high recovery percentage values (between 99% and 103%) and low relative standard deviation (RSD%) values (between 2% and 7%) for triolein and oleic acid standard solutions, as well as appreciably low limit of detection (LOD) and limit of quantification (LOQ) values (respectively 58 and 177 ng mL-1 for triolein; 198 and 602 ng mL-1 for oleic acid). Finally, the developed HPLC-ELSD method was successfully applied to evaluate the inhibitory effect of a polyphenolic extract obtained from apple pomace. The results showed a comparable inhibition degree between a 4.0mg mL-1 apple pomace solution and a 1.0μg mL-1 Orlistat solution. The proposed innovative method reveals highly sensitive and simple to follow the fate of PL digestion, thus opening the way to further investigations in the research of new potentially anti-obesity compounds. © 2022 Society of Chemical Industry.

Green, Rapid, Robust Determination of Aspartame in Beverages by High-Performance Liquid Chromatography-Evaporative Light Scattering Detection (HPLC-ELSD): Method Development and Validation

In recent years, eco-friendly approaches and reduction in the use of hazardous solvents have become important in applications of routine high-performance liquid chromatography (HPLC). Despite its various health side effects, aspartame (ASP) is a widely used artificial sweetener due to its high sweetness and low cost. In this study, a green, eco-friendly, cost effective, rapid, robust, and simple high-performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) method has been developed and validated for the determination of ASP in beverages. Separation was conducted isocratically on a C18 (4.6 × 150 mm, 3.5 µm) column; the mobile phase consisted of 80:20 water:ethanol at a flow rate of 1 mL min−1. The drift tube of evaporative light scattering detection (ELSD) temperature was adjusted to 50 °C and the nitrogen pressure was adjusted to 350 kPa. The HPLC-ELSD method provided a limit of detection and a limit of quantification of 0.33–0.99 µg mL−1, with recoveries of 99.4 ± 0.68% and 102.8 ± 0.17%, respectively. The precision of the method as the relative standard deviation was less than 1%. The method permitted the determination of ASP within 8 min using only 6.4 mL water and 1.6 mL ethanol. This study shows that the developed HPLC-ELSD method is suitable for determination of ASP in the food industry.

Antineoplastic and Antitrypanosomal Properties of Propolis from Tetragonula biroi Friese.

Propolis, popularly known as bee glue, is a resinous, sticky substance produced by different bee species across the globe. Studies on the biological properties of propolis from the Philippines are rare. Hence, the current study aims at the chemical characterization of propolis produced by the stingless bees Tetragonula biroi Friese from the Philippines and to investigate its antitrypanosomal and anticancer properties. The determination of the chemical composition and characterization of propolis samples was achieved using liquid chromatography-mass spectrometry (LC-MS), -high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD), and nuclear magnetic resonance (NMR) spectroscopy. Three major triterpenes were isolated and identified using HRESI-MS and 1H/13C NMR techniques. The spectral studies confirmed the presence of compounds such as isomangiferolic acid, 27-hydoxymangiferonic acid, and 27-hydroxyisomangiferolic acid. All crude propolis samples, isolated fractions, and pure compounds demonstrated moderate antitrypanosomal and anticancer properties compared to control drugs. Amongst the tested compounds, 27-hydoxymangiferonic acid exhibited the highest antitrypanosomal activity at a concentration of 11.6 µg/mL. The highest anticancer effect was demonstrated by the Ph-2 fraction, followed by 27-hydroxyisomangiferolic acid, with IC50 values of 129.6 and 153.3 µg/mL. Thus, it can be concluded that the observed biological activity of Philippine propolis is due to the combinatorial effect or synergistic action of the active compounds 27-hydoxymangiferonic acid and 27-hydroxyisomangiferolic acid.

Determination of the Sugar Content in High-Sugar Beverages.

We determined the sugar content of 75 samples, including eight black sugar beverages (BSBs) and seven Dalgona lattes (DLs) from Seoul, Korea, using high-performance liquid chromatography-evaporative light scattering detector. The mean sugar content per 100 g was 12.7 g and 12.3 g for BSB and DL, respectively. According to the World Health Organization, the mean sugar content per serving in BSB and DL were 34.8 g and 32.5 g, respectively, equivalent to 65∼70% of the recommended daily intake for added sugars. The highest sugar was sucrose (56.5% in BSB and 64.4% in DL). The sugar content could be adjusted in 27 out of 40 stores for BSB and 25 out of 35 stores for DL. The relative standard deviation of the content (g/100 g) per store by the manufacturer ranged between 5.7∼21.6% for BSB and 2.3∼29.5% for DL. This suggests that a uniform recipe is required for manufacturing beverages.

Fabrication, characterization, and purification of nutraceutical diacylglycerol components from Camellia oil.

Converting triacylgycerols (TAGs) from edible oils and fats into structured diacylglycerols (DAGs) is meaningful for reducing obesity. Camellia oil, rich in linoleic acid, has the potential to form structured linoleic acid-1,3-diacylglycerol (LA-1,3-DAG) nutrients in the industry. In this research, the physicochemical properties of modified Camellia oil (MCO) by enzymatic esterification were analyzed by Gas Chromatography-Mass Spectrometry (GC-MS), Differential Scanning Calorimetry (DSC), High Performance Liquid Chromatography-Evaporative Light Scattering Detection (HPLC-ELSD), and Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS). The relationship between reaction conditions and the DAG compositions is disclosed using multiple factors. It is found that high constituents of DAG increase the melting and crystallization temperature of MCO, lipase Novozym 435 gives the best yield of targeted nutrients (DAG, 1,3-DAG, LA-DAG), and the mixture of lipases, Lipozyme TL IM and Lipozyme RM IM, shows a synergistic effect in the synthetic process of DAG. Subsequently, MCO containing 65.4% DAG, 54.7% LA-DAG, and 47.6% 1,3-DAG content at optimal conditions (2% enzyme dosage, 4 h reaction time, 2.4:1 substrate molar ratio, 25.8% t-butanol as solvent, 60°C temperature) has been obtained and purified using silica column to obtain the final DAG oil containing 96.1% DAG, 64.7% 1,3-DAG, and 78.4% LA-DAG. High constituents of structured DAG oil rich in LA-1,3-DAG can be obtained by enzymatic esterification for industrial production.

Structure‐guided preparation of fuctional oil rich in 1,3‐diacylglycerols and linoleic acid from Camellia oil by combi‐lipase

Diacylglycerol (DAG)-enriched oil has been attracting attention because of its nutritional benefits and biological functions, although the composition of its various free fatty acids (FFAs) and an unclear relationship between substrate and yield make it difficult to be identified and qualified with respect to its production. In the present study, linoleic acid-enriched diacylglycerol (LA-DAG) was synthesized and enriched from Camellia oil by the esterification process using the combi-lipase Lipozyme TL IM/RM IM system. The relationship between FFA composition and DAG species productivity was revealed. The results showed that heterogeneous FFA with a major constituent (more than 50%) exhibited higher DAG productivity and inhibited triacylglycerol productivity compared to homogeneous constituents. Joint characterization by high-performance liquid chromatography-evaporative light scattering detection, gas chromatography-mass spectrometry and ultra-performance liquid chromatography-heated electrospray ionization-tandem mass spectrometry identified that DAG components contained dilinoleic acid acyl glyceride, linoleyl-oleyl glyceride and dioleic acid acyl glyceride in esterification products. Under the optimum conditions, 60.4% 1,3-DAG and 61.3% LA-DAG in the crude product at 1h reaction were obtained, and further purified to 81.7% LA-DAG and 94.7% DAG via silica column chromatography. The present study provides a guideline for the identification of DAG species, as well as a structure-guided preparation method of DAG-enriched oils via the cost-effective combi-lipase. © 2022 Society of Chemical Industry.

Effects of different processing methods on the content of astragaloside in danggui buxue decoction detection by high performance liquid chromatography-evaporative light scattering detection

Danggui Buxue decoction were prepared by different processing methods of angelica sinensis and astragalus membranaceus in wine and in oil respectively. High performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) method was established for the determination of astragaloside in Danggui Buxue decoction prepared by two processing methods, and the method was verified. The regression equations of astragaloside prepared by wine was y = 1412179x − 165809, R2 = 0.99827, in the range of 0.216~0.864 mg/mL; while in oil the regression equations was y = 1283633x − 141049, R2 = 0.9947, in the range of 0.162~0.648 mg/mL. It indicated that the peak area and concentration of astragaloside showed a good linear relationship, which met the requirements of detection. The average recovery of astragaloside prepared by wine was 94.5% and RSD was 0.763%, while they were 93.6% and 2.57% astragaloside prepared by oil. It shows that the accuracy of the method is good. RSD of 12 samples from the two individuals ranged from 1.67% to 1.38% in astragaloside prepared by wine, while it ranged from 3.34% to 3.33% in astragaloside prepared by oil, which proved good intermediate precision using HPLC-ELSD method. The RSD of peak area of astragaloside of the two products were all <2% within 12 h, indicating that the two samples were stable within 12 h. The content of astragaloside prepared by wine was 0.02514±0.00026 mg/mL. The content of astragaloside prepared by oil was 0.01949±0.00023 mg/mL. The results show that the sensitivity, precision and stability of this method are good, which are accurate and accord with the methodological verification. In addition, the processing methods of angelica sinensis and astragalus membranaceus in wine are more suitable for preparing Danggui Buxue decoction.

A high-performance liquid chromatography-evaporative light scattering detection-based quantitative analytical method for polyvinyl alcohol in food supplements with pyrolysis-gas chromatography/mass spectrometry-based verification.

Polyvinyl alcohol (PVOH) is extensively used in agricultural, pharmaceutical, textile, and food industries. A new high-performance liquid chromatography (HPLC) method with evaporative light scattering detection (ELSD) was developed for the quantitative analysis of PVOH products. Pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) was employed for method verification. The HPLC-ELSD method exhibited acceptable linearity (R2 > 0.99), with limits of detection and quantitation of 19.43 and 58.88μg/mL, respectively. Accuracies of 91.16-102.30% were estimated based on recoveries of the intra- and inter-day tests of PVOH. Repeatability and intermediate precision (%RSD) of 1.23-4.45 and 2.18-6.95, respectively, were obtained. The presence of PVOH in the HPLC peaks was further verified using typical indicators of PVOH in Py-GC/MS analysis, such as acetaldehyde, 2,5-dihydrofuran, benzaldehyde, and crotonaldehyde. This novel HPLC method with Py-GC/MS-based verification can be successfully applied for analyzing PVOH in dietary supplement tablets.

Construction of a high affinity aptamer and an aptasensor with chitosan oligosaccharide-AuNPs@Fe2+ nanozyme for highly sensitive detection of phosphatidylserine

In this study, a post-selection strategy based on increasing key bases was proposed to improve aptamers’ binding affinity. Phosphatidylserine (PS) was targeted as a model. Key bases of the original anti-PS aptamer were identified based on the molecular docking study, taken as repeating motif, and grafted onto the aptamer, with expectation to enhance the binding ability of the aptamer. An aptamer named PS-Apt-RKB2 was generated with significantly enhanced affinity. Biolayer interferometry assay results showed that the PS-Apt-RKB2 (Kd= 220.6 nM) exhibited a 29.59 times higher affinity over the original anti-PS aptamer (Kd= 6528 nM). Then, the PS-Apt-RKB2 was combined with chitosan oligosaccharide (COS) and AuNPs@Fe2+ to construct a label-free and COS-AuNPs@Fe2+ nanozyme-based colorimetric aptasensor. Under optimum conditions, the established aptasensor showed high sensitivity and demonstrated great application potentials in practice, with a limit of detection as low as 44.46 pM, and comparison rates of 91.21–105.4% when analyzing PS in real-world bioconversion samples with concentrations verified by high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD). This study can be further referred for developing well-performed aptamers and aptasensors for other targets.

Method Validation and Rapid Determination of Monosodium Glutamate in Various Food Products by HPLC-Fluorescence Detection and Method Optimization of HPLC-Evaporative Light Scattering Detection Approach without Derivatization.

In this study, an effective, simple and rapid high-performance liquid chromatography (HPLC) using fluorescence (FLD) method was developed and validated for the determination of monosodium glutamate (MSG) in 57 various food samples. Besides, HPLC-Evaporate Light Scattering Detection (ELSD) method was carried out for determination of MSG without derivatization. MSG analysis was performed by derivatization with dansyl chloride at excitation 328, emission 530nm with fluorescence detector. HPLC-FLD method was carried out by using C18 (150mm, 4.6mm, 2.7μm) column with the mobile phase consisting of (Water: Methanol:Glacial Acetic Acid)/(54:45:1,v/v/v). The column temperature was set at 25°C and the flow rate was set at 0.5mLmin-1 with an injection volume 20μL. The results were linear (R2 = 0.9999) with very low quantification limits. The applied method was optimized and the validated parameters such as LOD, LOQ, accuracy, precision, linearity and robustness were calculated. The obtained results were statistically compared with each other. The validated HPLC-FLD method was successfully applied for the analysis of MSG in all of the food samples. Moreover, HPLC-ELSD method was optimized and successfully demonstrated for detect the MSG without derivatization.

Development of methods for determining free fatty acid contents in red colored oils.

The online version contains supplementary material available at 10.1007/s10068-021-00964-2.

Seed oil of Brucea javanica induces apoptosis through the PI3K/Akt signaling pathway in acute lymphocytic leukemia Jurkat cells.

Brucea javanica oil emulsion (BJOE) has been used to treat tumor in China for more than 40 years. However, its components and effectiveness in the treatment of acute lymphocytic leukemia (ALL) and its mechanism of anti-cancer activity remain unknown. In the current study, high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) was used to analyze the components of BJOE. Then, the anti-leukemia effects of BJOE were examined both in vitro and in vivo using ALL Jurkat cells and the p388 mouse leukemia transplant model, respectively. The primary ALL leukemia cells were also used to confirm the anti-leukemia effects of BJOE. The apoptotic-related results indicated that BJOE induced apoptosis in Jurkat cells and were suggestive of intrinsic apoptotic induction. Moreover, BJOE inhibited Akt (protein kinase B) activation and upregulated its downstream targets p53 and FoxO1 (forkhead box gene, group O-1) to initiate apoptosis. The activation of GSK3β was also involved. Our findings demonstrate that BJOE has anti-leukemia effects on ALL cells and can induce apoptosis in Jurkat cells through the phosphoinositide3-kinase (PI3K) /Akt signaling pathway.

Lipase-Catalyzed Production of Sorbitol Laurate in a "2-in-1" Deep Eutectic System: Factors Affecting the Synthesis and Scalability.

Surfactants, such as glycolipids, are specialty compounds that can be encountered daily in cleaning agents, pharmaceuticals or even in food. Due to their wide range of applications and, more notably, their presence in hygiene products, the demand is continuously increasing worldwide. The established chemical synthesis of glycolipids presents several disadvantages, such as lack of specificity and selectivity. Moreover, the solubility of polyols, such as sugars or sugar alcohols, in organic solvents is rather low. The enzymatic synthesis of these compounds is, however, possible in nearly water-free media using inexpensive and renewable building blocks. Using lipases, ester formation can be achieved under mild conditions. We propose, herein, a “2-in-1” system that overcomes solubility problems, as a Deep Eutectic System (DES) made of sorbitol and choline chloride replaces either a purely organic or aqueous medium. For the first time, 16 commercially available lipase formulations were compared, and the factors affecting the conversion were investigated to optimize this process, owing to a newly developed High-Performance Liquid Chromatography-Evaporative Light Scattering Detector (HPLC-ELSD) method for quantification. Thus, using 50 g/L of lipase formulation Novozym 435® at 50 °C, the optimized synthesis of sorbitol laurate (SL) allowed to achieve 28% molar conversion of 0.5 M of vinyl laurate to its sugar alcohol monoester when the DES contained 5 wt.% water. After 48h, the de novo synthesized glycolipid was separated from the media by liquid–liquid extraction, purified by flash-chromatography and characterized thoroughly by one- and two-dimensional Nuclear Magnetic Resonance (NMR) experiments combined to Mass Spectrometry (MS). In completion, we provide initial proof of scalability for this process. Using a 2.5 L stirred tank reactor (STR) allowed a batch production reaching 25 g/L in a highly viscous two-phase system.

Determination of Polyvinyl Acetate in Chewing Gum Using High-Performance Liquid Chromatography-Evaporative Light Scattering Detector and Pyrolyzer-Gas Chromatography-Mass Spectrometry.

Polyvinyl acetate (PVAc) is used in various adhesive, paint, and transparent tape applications. It is also used as a food additive in food manufacturing to make chewing gum and fruit and vegetable glazes; however, guidelines on the amount of food additives that is used have not yet been established. In this study, a method was developed for analysis of polyvinyl acetate (PVAc) using high-performance liquid chromatography with an evaporative light scattering detector (HPLC–ELSD) and pyrolyzer–gas chromatography–mass spectrometry (Py–GC–MS). The analytical methods were applied to commercially available chewing gum. In the HPLC–ELSD analysis, the linearity was acceptable (R2 > 0.999), and the limits of detection and quantification were 22.2 and 67.3 µg/mL, respectively. The accuracies of PVAc were 87–115% at spike levels of 200–1000 µg/mL for the intra- and inter-day tests. The contents of PVAc in the chewing gum samples were n.d. (not detected)—13.8 g/kg. The presence of PVAc in chewing gum was verified with Py–GC–MS analysis, finding the typical pyrolysates of PVAc, such as acetic acid, benzene, toluene, styrene, indane, naphthalene, and acenaphthene. The developed analytical methods can be applied for successful identification of PVAc in chewing gum.