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FOOD CHEMISTRY · 2024 · IF 8.5

Comparative study on the enzymatic degradation of phenolic esters: The HPLC-UV quantification of tyrosol and gallic acid liberated from tyrosol acyl esters and alkyl gallates by hydrolytic enzymes

Xinmiao Wang,Qian Wang,Dong Cai,Jinghan Yu,Xuan Chen,Xu Guo,Peiyong Tong,Xiaoyang Liu,Fawen Yin,Dayong Zhou

HPLC-UV analysis was used to evaluate the enzymatic degradation characteristics of tyrosol acyl esters (TYr-Es) and alkyl gallates (A-GAs). Among various hydrolytic enzymes, TYr-Es can be hydrolyzed by pancrelipase, while A-GAs cannot be hydrolyzed by pancrelipase. Interestingly, carboxylesterase-1b (CES-1b), carboxylesterase-1c (CES-1c) and carboxylesterase-2 (CES-2) are able to hydrolyze TYr-Es and A-GAs, and thus to liberate tyrosol (TYr) and gallic acid (GA). By contrast, the degrees of hydrolysis (DHs) of TYr-Es and A-GAs by CES-1b and CES-1c were significantly higher than those by CES-2. Meanwhile, the DHs of TYr-Es were much higher than those of A-GAs. Especially, the DHs firstly increased and then decreased with the increasing alkyl chain length. Besides, DHs positively correlated with the unsaturation degree at the same chain length. Through regulating carbon length, unsaturation degree and the ester bond structure, controlled-release of phenolic compounds and fatty acids (or fatty alcohols) from phenolic esters will be easily achieved.

研究领域: 生物化学 · 食品科学
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INDUSTRIAL CROPS AND PRODUCTS · 2024 · IF 5.6

Selection of moderate refining process for gardenia fruit oil based on SPE-HPLC-UV analysis of phytonutrients

Weikang Shen,Haobo Wang,Zhouzhe Lan,Guangzhi Xu,Qinxue Ni,Qiufen Mo,Youzuo Zhang,Yan Wang

Gardenia fruit oil (GFO) is a new emerging edible woody oil with few investigations of its’ refinement process. In this study, different degumming, deacidification and bleaching technologies were introduced for realizing moderate refining of the GFO based on the preservation of bioactive micronutrients. Firstly, a simple multi-stage solid phase extraction (SPE) combined with a lab- affordable HPLC-UV instrument was developed for determinations of several vital biological constituents, such as crocetin, geniposide, polyphenols, phytosterols and squalene in the GFO. Satisfactory limit of detection (LOD), limit of quantification (LOQ), recovery, intra-day and inter-day precision were obtained. Subsequently, changes of those phytochemicals during three types of degumming, followed by three kinds of deacidification and then three ways of bleaching were investigated, offering insightful suggestions for achieving moderate refining of the GFO. Finally, the refined GFO might be suggested as a potential functional edible oil with valuable micronutrients. To our knowledge, this is the first study investigating moderate refinement techniques for GFO, as well as developing novel methods for phytonutrients analysis in the GFO.

研究领域: 食品科学 · 分析化学 · 植物化学
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Analytical Methods · 2018 ·Vol. 10 ·期 11 ·1325-1330 · IF 2.073

Fast simultaneous detection of three diterpenoids in Herba Siegesbeckiae using solid phase extraction followed by HPLC-UV with a core–shell particle column

Yong Zhang,Zhizhong Zou,Guixing Chou

To quickly determine the amounts of kirenol, darutoside and darutigenol in Herba Siegesbeckiae using HPLC, the efficiency of sample cleanup with different solid phase extraction methods was systematically investigated. The results showed that a polymer based strong anion exchange (P-SAX, 200 mg/6 mL) sorbent could separate and enrich diterpenoids providing rapid determination without impurity interference. In this paper a new HPLC-UV method using a core–shell column (Boltimate C18 3.0 × 100 mm, 2.7 μm) for the quantitative determination of three diterpenoids was developed. Under optimum experimental conditions, P-SAX pretreatment with a core–shell analytical column exhibited satisfactory results in the determination of diterpenoids in Herba Siegesbeckiae with recovery rates of 95–102%, relative standard deviations below 1.1%, a limit of quantitation as low as 0.1 6 μg mL−1, and a short analytical runtime of 8 min. Finally, the proposed method was successfully applied in the profiling of diterpenoids in Herba Siegesbeckiae from different regions.

CAS / MDL: 7664-38-2
研究领域: 分析化学 · 生药学 · 植物科学
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Heliyon · 2023 · IF 4

Development and validation of a novel HPLC-UV method for simultaneous determination of azathioprine metabolites in human red blood cells

Hengyi Yu,Dongyan Li,Dong Xiang,Xiping Li,Lu Liu,Dong Liu,Xuepeng Gong

A rapid, specific and accurate high-performance liquid chromatography with tunable ultraviolet detection method was developed to simultaneously determine azathioprine metabolites, 6-thioguanine nucleotides (6-TGN) and 6-methyl mercaptopurine riboside (6-MMPr) in human red blood cells. Erythrocyte lysate sample was precipitated by perchloric acid under the protection of dithiothreitol, with 6-TGN and 6-MMPr being acid hydrolyzed to produce 6-thioguanine (6-TG) and 6-methymercaptopurine (6-MMP). A Waters Cortecs C 18 column (2.1 × 150 mm, 2.7 μm) was used for chromatographic separation with a water (containing 0.01 mol/L ammonium acetate and 0.2% acetic acid)/methanol linear gradient at a flow rate of 0.45 mL/min in a 5.5 min. UV detection wavelengths were 340 nm for 6-TG, 303 nm for 6-MMP and the IS (5-bromouracil). The calibration curves fitted a least squares model (weighed 1/ x 2 ) from 0.15 to 15 μmol/L for 6-TG ( r 2  = 0.9999) and from 1 to 100 μmol/L for 6-MMP ( r 2  = 0.9998). This method was validated according to the FDA bioanalytical method validation guidance and ICH M10 bioanalytical method validation and study sample analysis guidance for industry, and successfully utilized in ten IBD patients receiving azathioprine therapy.

研究领域: 药理学 · 分析化学 · 临床化学
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Talanta Open · 2025 · IF 3.7

Quantification of oleanolic acid and apigenin in biological matrices following intraperitoneal administration of co-loaded nanoformulations: Method validation and biodistribution studies

Amna Naz,Sadia Haider,Mavra Rana,Neelum Malik,Attia Afzal,Muhammad Farooq,Muhammad Sarfraz

Oleanolic acid (OA) and apigenin (APG) demonstrate a wide range of pharmacological effects, including antioxidant, anti-inflammatory, antiviral, antidiabetic, hepatoprotective, and cardioprotective activities. Both compounds show anticancer potential and have been used individually to treat various cancer types, creating a rationale for their combined use. However, quantifying both drugs in quality and plasma samples presents challenges due to differences in their physicochemical properties and absorption wavelengths (OA: 210-215 nm, APG: 212, 269, and 337 nm). We developed and validated a novel High-Performance Liquid Chromatography UV detection (HPLC-UV) method to address this problem. Using a C18 column (4.6 mm x 250 mm, packing L1, 5 µm particle size) and a mobile phase of acetonitrile: methanol: tetrahydrofuran: water (60:20:8:12, v/v) with a detection wavelength of 215 nm. The method achieved retention times of 4.7 and 10 minutes for OA and APG in quality samples, and 4.9 and 9.8 minutes in plasma samples, respectively. The plasma sample was prepared by simple protein precipitation step. All validation parameters, including sensitivity, selectivity, precision, accuracy, robustness, and stability, met the required criteria. The validated HPLC-UV bioanalytical method was effectively applied in quantification, stability, and pharmacokinetic studies of co-loaded nanoformulations of OA and APG with particle size 163 nm and entrapment efficiency up to 93 %, establishing it as a promising analytical technique for detecting these anticancer agents in complex matrices such as blood, plasma, and polymeric systems.

CAS / MDL: 520-36-5
研究领域: 药理学 · 分析化学 · 肿瘤学
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SEPARATION AND PURIFICATION TECHNOLOGY · 2015 · IF 3.091

Preparation and determination of phenolic compounds from Pyrola incarnata Fisch. with a green polyols based-deep eutectic solvent

Xiao-Hui Yao,Dong-Yang Zhang,Ming-Hui Duan,Qi Cui,Wen-Jing Xu,Meng Luo,Chun-Ying Li,Yuan-Gang Zu,Yu-Jie Fu

A group of green solvent named polyols-based deep eutectic solvents (PDES) combined with microwave assisted extraction (MAE) was first successfully applied in extracting phenolic compounds from pyrola. A series of PDES with mixing different ratios choline chloride (ChCl) to different polyols were investigated for extraction, and ChCl/1,4-Butanediol (1/4) was found to be the suitable extraction solvent. MAE was more effective than heat-stirring extraction and ultrasonic assisted extraction with PDES. The extraction parameters were optimized by BBD test combined with response surface methodology. The optimal extraction conditions were 30% of water in ChCl/1,4-Butanediol (1/4) at 70   °C for 20   min and liquid/solid ratio 10   mL/g. Moreover, RP-HPLC-UV was developed for the determination of five main active components in pyrola. This method exhibited satisfactory linearity ( R 2   >   0.999), precision (RSD   <   4.03%) and recovery (96.17–103.25%). Sample analysis showed that the contents of hyperin, 2′-O-galloylhyperin, quercitrin, quercetin-O-rhamnoside and chimaphilin in pyrola were 1.627, 4.958, 0.041, 0.089 and 0.349   mg/g, respectively. These results revealed that PDES would be promising green solvents for analytical sample preparation from plant herbs.

研究领域: 植物化学 · 分析化学
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JOURNAL OF FOOD COMPOSITION AND ANALYSIS · 2024 · IF 4

Cation-exchange/hydrophilic interaction mixed-mode liquid chromatographic method for analyzing the trigonelline class of bioactives in coffee beans

Jian-Jun Zhong,Lingling Zhao,Yuanxin Guo,Zhanming Li,Hua Ye,Huanan Guan,Zhiqiang Xing,Weiqing Wang

The coffee phytochemical trigonelline and its roasting products nicotinic acid and N -methylpyridinium represent an important class of bioactives that mediate the health benefits of coffee consumption. However, there is a paucity of methods for their simultaneous analysis in coffee beans, especially those affordable and reliable for widespread routine use. To improve this situation, an easy-to-use HPLC−UV method was developed in this study based on a reversed-phase/cation-exchange mixed-mode column showing unique chromatographic selectivity toward the polar and ionic analytes over the matrix components of coffee. The selectivity was achieved by using a decreasing acetonitrile gradient, which tuned the column to a cation-exchange/hydrophilic interaction (CEX/HILIC) mixed separation mode whilst minimizing its hydrophobic interaction capability. The method was assessed for figures of merit, giving limits of detection in coffee beans at around the 0.1 mg/100 g wet matter level, and verified for its applicability to a heterogeneous set of coffee bean samples differing in roasting degree and origin.

研究领域: 分析化学 · 食品科学 · 植物化学
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Food Analytical Methods · 2020 ·Vol. 13 ·期 8 ·1673-1680 · IF 2.667

Rapid Determination for Benzoic Acid, Sorbic Acid, Phenyllactic Acid, Phenylalanine, and Saccharin Sodium in Vinegar by High-Performance Liquid Chromatography–UV

Cheng Zhu,Ran Qin,Liu Jie,Deng Xing,Qiu Huidong,Jia Zhengfu,Su Xiaodong

Preservatives and sweeteners are always added in the food production process, which can extend the life and flavor of food. Benzoic acid and saccharin sodium are commonly used as preservatives and sweeteners respectively. Phenyllactic acid as a natural preservative, mainly produced by phenylalanine fermentation, may exist in fermented food. The objective of the current study is to develop a reverse-phase HPLC-UV method for simultaneous determination of benzoic acid, sorbic acid, phenyllactic, phenylalanine, and saccharin sodium in vinegar. They were separated on Sapphiresil TM (C 18 ) column (250 × 4.6 mm, 5 μm) with a mobile phase consisting of methyl alcohol:ammonium acetate buffer (pH 6.44; 0.02 M) (20:80, v/v) ran in isocratic mode at a flow rate of 1 mL/min. Complete baseline separation of five analytes was achieved in < 12 min, the linear range was 0.25–50 μg/mL with a correlation coefficient R 2 value > 0.9975, the method was validated with recoveries in the 96.93 to 118.42% range, and RSD was 0.39–6.21%. The detection limits of phenylalanine, benzoic acid, saccharin sodium, sorbic acid, and phenyllactic acid were 0.25, 0.01, 0.01, 0.025, and 0.05 mg/L. This method could be used for determining of benzoic acid, sorbic acid, phenyllactic acid, phenylalanine, and saccharin sodium in vinegar. Graphical abstract

研究领域: 食品科学 · 分析化学 · 食品安全
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CHEMOSPHERE · 2018 · IF 4.427

Applying molecular modelling and experimental studies to develop molecularly imprinted polymer for domoic acid enrichment from both seawater and shellfish

Junjie Ao,Jiaping Gu,Tao Yuan,Dan Li,Yuning Ma,Zhemin Shen

A highly selective sample cleanup method using molecularly imprinted polymers (MIP) was developed for the enrichment of domoic acid (DA, an amnesic shellfish toxin) from both seawater and shellfish samples. Molecular modelling was firstly applied to screening a suitable functional monomer and optimize the polymer preparation. Theoretical results were in a good agreement with those of the experimental studies. MIP was prepared by precipitation polymerization using 1, 3, 5-pentanetricarboxylic acid and 2-(Trifluoromethyl)acrylic acid as the template molecule and functional monomer, respectively. The morphology and molecular structure of MIP were revealed by scanning electron microscope (SEM) and fourier transform infrared spectroscopy (FTIR), respectively. The obtained MIP showed high affinity and selectivity for DA with binding site numbers of 0.875 mg g −1 and an average association constant of 0.219 L mg −1 evaluated by adsorption experiments. The developed molecularly imprinted solid-phase extraction (MISPE) column achieved satisfied adsorption rate (99.2%) and recovery (71.2%) with relative standard deviation (RSD) less than 1.0%, which is more stable and precise than the C 18 , SAX, and HLB columns. Finally, the determination method for DA in both seawater and shellfish samples was then successfully established and validated using MISPE coupled with high-performance liquid chromatography-ultraviolet detection (HPLC-UV). The method limit of detection was 20 μg L −1 and 50 μg kg −1 for seawater and shellfish, respectively. This study demonstrates that molecular modelling is a useful tool to screening functional monomer and optimize polymer preparation. It provides an innovative polymer for trace DA monitoring in both seawater and shellfish.

研究领域: 分析化学 · 环境科学 · 海洋生物学 · 毒理学
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Analytical Methods · 2014 ·Vol. 6 ·期 5 ·1563-1568 · IF 1.938

Pharmacokinetic study of liquiritin in rat serum using molecularly imprinted solid-phase extraction combined with RP-HPLC

Huang Yang,Li Kang,Shao Hui Kai,Liu Sheng Quan,Xiang Fei Jun

A method using molecularly imprinted solid-phase extraction combined with RP-HPLC-UV was developed for determination of liquiritin (LQR) in rat plasma after oral administration of Compound Liquorice Tablets (CLTs). To this end, molecularly imprinted polymers were prepared by in situ polymerization, using LQR as the template, acrylamide as the functional monomer, and ethylene glycol dimethacrylate as the crosslinker, whose solid-phase extraction properties for liquiritin were evaluated by HPLC-UV. The enrichment and extraction recovery were investigated in the study after the extraction procedure was optimized. The recovery was larger than 85.0% and the RSD was less than 10.0%. The calibration curve was expressed as y = 16 004x − 1.864 (r = 0.996) with a linear range of 0.007–1.683 μg mL−1. Besides, the Tmax was 0.5 h for liquiritin and the Cmax was 0.8 μg mL−1 after oral administration of compound liquorice tablets. In addition, the pharmacokinetic parameters were calculated using a two-compartment model. The results indicated that the monolithic polymer pipette presented a good extraction efficiency for liquiritin. At the same time, the results indicated that the index components could basically reveal the pharmacokinetic behavior. This study was aimed to explore an approach that could be applied to promulgate the pharmacokinetics and provide more information for clinical applications.

CAS / MDL: 79-06-1 2867-47-2
研究领域: 药理学 · 分析化学 · 生物医学工程
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BIOTECHNOLOGY LETTERS · 2017 ·Vol. 39 ·期 8 ·1229-1235 · IF 1.73

Molecular cloning and expression of a glycosyltransferase from Bacillus subtilis for modification of morin and related polyphenols

Wang Qianqian,Xu Yixiang,Xu Jiaqi,Wang Xudong,Shen Chen,Zhang Yan,Liu Xiufeng,Yu Boyang,Zhang Jian

Objectives To characterize glycosyltransferases from Bacillus subtilis ATCC 6633 and investigate their substrate specificity towards plant polyphenols. Results Among the cloned and expressed six UDP-glycosyltransferases (BsGT1-6), BsGT-1 showed activity with a wide range of polyphenols: morin, quercetin, alizarin, rehin, curcumin and aloe emodin. The gene of BsGT-1 has an ORF of 1206 bp encoding 402 amino acids. The recombinant enzyme was purified to homogeneity by Ni–NTA affinity chromatograph, and its biochemical characteristics were identified by HPLC–UV/MS, 1 H-NMR and 13 C-NMR. BsGT-1 has an MW of approx. 46 kDa as indicated by SDS-PAGE; its activity was optimal at 40 °C and pH 8.5. The Km value of BsGT-1 towards morin was 110 μM. Conclusions BsGT-1 from B. subtilis was cloned. It had high catalytic capabilities towards polyphenols which would make it feasible for the structural modification of polyphenols.

研究领域: 生物化学 · 分子生物学 · 酶学
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BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY · 2023 ·Vol. 70 ·期 6 ·1806-1816 · IF 2.8

Quantitative determination of rhamnolipid using HPLC-UV through carboxyl labeling

Jing Zhou,Si-Jia Miao,Shi-Zhong Yang,Jin-Feng Liu,Hong-Ze Gang,Bo-Zhong Mu

Rhamnolipid, as a low-toxic, biodegradable and environmentally friendly biosurfactant, has broad application prospects in many industries. However, the quantitative determination of rhamnolipid is still a challenging task. Here, a new sensitive method for the quantitative analysis of rhamnolipid based on a simple derivatization reaction was developed. In this study, 3-[3′-( l -rhamnopyranosyloxy) decanoyloxy] decanoic acid (Rha-C10-C10) and 3-[3′-(2′- O -α- l -rhamnopyranosyloxy) decanoyloxy] decanoic acid (Rha-Rha-C10-C10) were utilized as the representative rhamnolipids. Liquid chromatography–mass spectrometry and high-performance liquid chromatography-ultra violet results showed that these two compounds were successfully labeled with 1 N1-(4-nitrophenyl)-1,2-ethylenediamine. There was an excellent linear relationship between rhamnolipid concentration and peak area of labeled rhamnolipid. The detection limits of the Rha-C10-C10 and Rha-Rha-C10-C10 were 0.018 mg/L (36 nmol/L) and 0.014 mg/L (22 nmol/L), respectively. The established amidation method was suitable for the accurate analysis of rhamnolipids in the biotechnological process. The method had good reproducibility with the relative standard deviation of 0.96% and 0.79%, respectively, and sufficient accuracy with a recovery of 96%–100%. This method was applied to quantitative analysis of 10 rhamnolipid homologs metabolized by Pseudomonas aeruginosa LJ-8. The single labeling method was used for the quantitative analysis of multiple components, which provided an effective method for the quality evaluation of other glycolipids with carboxyl groups.

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