Comparison of 24-h volume and creatinine-corrected total urinary polyphenol as a biomarker of total dietary polyphenols in the Invecchiare InCHIANTI study

Polyphenols have beneficial effects on several chronic diseases but assessing polyphenols intake from self-reported dietary questionnaires tends to be inaccurate and not very reliable. A promising alternative is to use urinary excretion of polyphenols as a proxy measure of intake. The best method to assess urinary excretion is to collect 24-h urine. However, since collecting 24-h urine method is expensive, time consuming and may be difficult to implement in large population-based studies, measures obtained from spot urine normalized by creatinine are commonly used. The purpose of the study was to evaluate the correlation between polyphenols dietary intake and total urinary polyphenol excretion (TPE), expressed by both 24-h volume and urinary creatinine normalization in 928 participants from the InCHIANTI study. Dietary intake data were collected using a validated food frequency questionnaire. Urinary TPE was analyzed by Folin-Ciocalteau assay. Both urinary TPE expression models were statistically correlated (r=0.580), and the partial correlation coefficient improved (pr=0.722) after adjusting for the variables that modify the urinary creatinine excretion (i.e. gender, age, BMI, physical activity and renal function). In crude models, polyphenol intake was associated with TPE corrected by 24-h volume (r=0.211; P<0.001), but not with creatinine normalization (r=0.014; P=0.692). However, urinary TPE expressed by creatinine correction was significantly correlated with dietary polyphenols after adjusting for covariates (pr=0.113; P=0.002). We conclude that urinary TPE expressed by 24-h volume is a better biomarker of polyphenol dietary intake than by urinary creatinine normalization. After covariate adjustment, both can be used for studying the relationships between polyphenol intake and health in large-scale epidemiological studies.     

高效阴离子交换色谱-脉冲安培检测法分析啤酒和麦汁中的糖

建立了高效阴离子交换色谱-脉冲安培检测(HPAEC-PAD)同时测定单糖、二糖和多种低聚糖的方法。采用水、0.25 mmol/L NaOH溶液和1 mol/L NaAc溶液三元梯度淋洗,在CarboPac PA-100色谱柱上,11种糖在40 min内达到良好分离;采用积分脉冲安培检测方法,无需对样品进行复杂的前处理或衍生反应便可直接检测。11种糖的检出限(S/N=3)在13～88 pg范围内。将该方法用于啤酒和麦汁样品中单糖、二糖及低聚糖的分析取得了很好的结果,样品中的加标回收率为81%～107%。     

Simultaneous assessment of cholesterol absorption and synthesis in humans using on-line gas chromatography/ combustion and gas chromatography/pyrolysis/isotope-ratio mass spectrometry.

A number of dietary components and drugs are known to inhibit the absorption of dietary and biliary cholesterol, but at the same time can compensate by increasing cholesterol synthesis. It is, therefore, necessary to have a convenient and accurate method to assess both parameters simultaneously. Hence, we validated such a method in humans using on-line gas chromatography(GC)/combustion and GC/pyrolysis/isotope-ratio mass spectrometry (IRMS). Cholesterol absorption was measured using the ratio of [(13)C]cholesterol (injected intravenously) to [(18)O]cholesterol (administered orally). Simultaneously, cholesterol synthesis was measured using the deuterium incorporation method. Our methodology was applied to 12 mildly hypercholesterolemic men that were given a diet providing 2685 +/- 178 Kcal/day (mean +/- SD) and 255 +/- 8 mg cholesterol per day. Cholesterol fractional synthesis rates ranged from 5.0 to 10.5% pool/day and averaged 7.36% +/- 1.78% pool/day (668 +/- 133 mg/day). Cholesterol absorption ranged from 36.5-79.9% with an average value of 50.8 +/- 15.4%. These values are in agreement with already known data obtained with mildly hypercholesterolemic Caucasian males placed on a diet similar to the one used for this study. However, our combined IRMS method has the advantage over existing methods that it enables simultaneous measurement of cholesterol absorption and synthesis in humans, and is therefore an important research tool for studying the impact of dietary treatments on cholesterol parameters.     

Hydrophilic interaction chromatography/electrospray mass spectrometry analysis of carbohydrate-related metabolites from Arabidopsis thaliana leaf tissue

This work describes the development and application of an on-line liquid chromatography/mass spectrometry (LC/MS) method using hydrophilic interaction chromatography (HILIC) coupled to negative ion mode electrospray ionisation ion trap mass spectrometry (ESI-MS) for the analysis of highly polar carbohydrate-related metabolites commonly found in plants, ranging from reducing and non-reducing sugars and sugar alcohols to sugar phosphates. Using this method, separation and detection of a mixture of eight authentic standard compounds containing glucose (Glc), sucrose (Suc), raffinose, verbascose, mannitol, maltitol, glucose-6-phosphate (Glc6P) and trehalose-6-phosphate (Tre6P) were achieved in less than 15 min. The method is rapid, robust, selective, and sensitive, with limits of detection (LODs) ranging from 0.2 microM obtained for neutral sugars, to 1.0 microM obtained for sugar alcohols, and 2.0 microM obtained for negatively charged sugar phosphates. We have studied the negative ion collision-induced dissociation (CID) fragmentation behaviour of the non-reducing raffinose family oligosaccharides (RFOs) raffinose, stachyose, and verbascose. Mainly Bi and Ci glycosidic and Ai cross-ring structurally informative cleavages are observed. We have applied this HILIC/ESI-MS method for the analysis of Arabidopsis thaliana wild-type Columbia-0 (Col-0) and its starchless phosphoglucomutase mutant (pgm1) leaf extracts. The method was used to quantify Glc, Suc, raffinose, and Glc6P in A. thaliana extracts. Data obtained using this HILIC/ESI-MS method were compared with those obtained using a comparable porous graphitic carbon-based LC/ESI-MS method.     

A new acetonitrile-free mobile phase method for LC-ELSD quantification of fructooligosaccharides in onion (Allium cepa L.)

Onion soluble non-structural carbohydrates consist of fructose, glucose and sucrose plus fructooligosaccharides (FOS) with degrees of polymerisation (DP) in the range of 3-19. In onion, sugars and FOS are typically separated using liquid chromatography (LC) with acetonitrile (ACN) as a mobile phase. In recent times, however, the production of ACN has diminished due, in part, to the current worldwide economic recession. A study was therefore undertaken, to find an alternative LC method to quantify sugars and FOS from onion without the need for ACN. Two mobile phases were compared; the first taken from a paper by Vågen and Slimestad (2008) [3] using ACN mobile phase, the second, a newly reported method using ethanol (EtOH). The EtOH mobile phase eluted similar concentrations of all FOS compared to the ACN mobile phase. In addition, limit of detection, limit of quantification and relative standard deviation values were sufficiently and consistently lower for all FOS using the EtOH mobile phase. The drawback of the EtOH mobile phase was mainly the inability to separate all individual sugar peaks, yet FOS could be successfully separated. However, using the same onion extract, a previously established LC method based on an isocratic water mobile phase could be used in a second run to separate sugars. Although the ACN mobile phase method is more convenient, in the current economic climate a method based on inexpensive and plentiful ethanol is a valid alternative and could potentially be applied to other fresh produce types.In addition to the mobile phase solvent, the effect of extraction solvents on sugar and FOS concentration was also investigated. EtOH is still widely used to extract sugars from onion although previous literature has concluded that MeOH is a superior solvent. For this reason, an EtOH-based extraction method was compared with a MeOH-based method to extract both sugars and FOS. The MeOH-based extraction method was more efficacious at extracting sugars and FOS from onion flesh, eluting significantly higher concentrations of glucose, kestose, nystose and DP5-DP8.     

Chemical profiling and characterization of phenolic acids,flavonoids, terpene glycosides from Vangueria agrestis using ultra-high-performance liquid chromatography/ion mobility quadrupole time-of-flight mass spectrometry and metabolomics approach

Vangueria agrestis is a shrub indigenous to tropical Africa, belonging to family Rubiaceae and is traditionally used as a decoction for treatment of fever, pain, and malaria. This study was undertaken to investigate the chemical constituents based on precursor exact mass and fragment ion information. The chemical profiling and structural characteristics of chemical constituents from methanolic extracts of dried aerial parts and roots of V. agrestis and dietary supplements were analyzed using ultra-high-performance liquid chromatography/ion mobility quadrupole time-of-flight mass spectrometry coupled with UNIFI platform and multivariate analysis in both negative and positive ion modes. A non-targeted ultra-high-performance liquid chromatography-mass spectrometry analysis was carried out to profile the chemical constituents of crude extracts of V. agrestis, and 73 compounds, including reference compounds, were identified. The fragments of flavonoids, monoterpene, and triterpene glycosides revealed the characteristic cleavage of glycosidic linkages, and the fragmentation pattern provided the identity of the sugars. This analytical method provides a quick method for quality assessment of dietary supplements. Finally, a chemometrics approach with multivariate statistical tools was used to visualize the differences between root and aerial parts of plant samples and to find the potential chemical markers that differentiate among these parts of V. agrestis samples and dietary supplements.     

Measurement of sugars and starches in foods by a modification of the AOAC total dietary fiber method.

A separation scheme for the determination of sugars and starch in processed food was developed. It is based on AOAC Method 985.29 for total dietary fiber with these modifications: carbohydrate starches are separated into soluble and insoluble fractions before they are hydrolyzed; acetonitrile is used instead of ethanol to separate sugars from enzyme-resistant carbohydrates, proteins, and other macromolecules; and a solid-phase extraction filter is included to remove substances that interfere with high-performance liquid chromatography (HPLC). Recovery studies indicate a > 97% sugar recovery. Twenty foods were analyzed. After enzymatic hydrolysis, fructose, glucose, sucrose, maltose, and lactose were extracted and determined by HPLC using a refractive index detector. Starch content was calculated from the increase in the amount of glucose. The results were compared with values listed on the "Nutrition Facts" panel for that food. The analyzed amounts of sugars and starches were 73-96% of declared values.     

Determination of carbohydrates in juices by capillary electrophoresis, high-performance liquid chromatography, and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry

The objective of this work was to develop a sample preparation procedure for determination of the carbohydrate profiles in commercial juice samples by three principally different analytical methods: capillary electrophoresis (CE) with indirect detection, high-performance liquid chromatography (HPLC), and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). The preparation and purification of juice samples prior to analysis is described. The method using Carrez reagents was found to be an efficient preparation tool for all three methods. The addition of Carrez reagents to the samples for mass analysis improved the quality of the mass spectra of oligosaccharides. The amounts of glucose, fructose, and sucrose as major carbohydrates in fruit juices measured by CE using a simple instrument are in good agreement with the HPLC values and the data declared by the producers of the juices. The results from both methods are critically evaluated and their impact for studies of authenticity is discussed. The decrease of sucrose amount during the storage of samples was explained by acid hydrolysis of this disaccharide.     

Quantification and validation of enzyme immunoassay for urinary aflatoxin B1-N7-guanine adduct for biological monitoring of aflatoxins

The aflatoxin B1-N7-guanine (AFB1-N7-guanine) adduct has been established as one of the relevant biomarkers of dietary aflatoxin (AFB1) exposure. Measurement of this adduct is potentially a useful dosimeter in molecular epidemiological studies. This paper reports the application and evaluation of a sensitive indirect competitive enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of urinary AFB1-N7-guanine adduct in high risk populations exposed to dietary aflatoxin. Earlier, we had reported a simple and rapid indirect ELISA method for AFB1-N7-guanine adduct in the urine and liver tissues using polyclonal antibodies specific to AFB1-N7-guanine adduct. The method was evaluated using a rodent model (Fischer 344), exposed to 1 mg kg-1 body mass of AFB1 and human urine samples obtained from a maize eating population, environmentally exposed to AFB1 through their diet. The levels of AFB1-N7-guanine adduct in rat and human urine ranged from 6.42 to 20.16 micrograms mg-1 creatinine and from 9.30 to 13.43 ng mg-1 creatinine, respectively. The level of AFB1 in the diet as estimated by ELISA ranged from 1000 to 3600 ng d-1. The interesting observation in these studies is that the females (in both rodents and human subjects) are more efficient than males at excreting the adduct. Total adduct (DNA bound adduct and guanine adduct excreted in urine) was found to be similar in male and female rats. However, 63% of the total adduct was accounted for in urine of female rats, whereas male rats excreted 47% of the total adduct in their urine. The present method may find wide application as a biochemical tool in molecular epidemiological studies with respect to human exposure to dietary aflatoxins.     

陈皮中农药残留分析及风险评估研究

通过分析中药陈皮的农药残留问题开展风险评估研究。采用气相色谱-串联质谱(GC-MS/MS)和高效液相色谱-串联质谱(HPLC-MS/MS)对198批陈皮中的117种农药进行检测;采用点评估方式计算陈皮中农药残留的急性和慢性摄入风险;采用英国兽药残留委员会提出的兽药残留风险排序矩阵计算各农药的风险得分;采用危害指数(HI)法计算有机磷农药的慢性累积风险。198批陈皮中共检出30种农药(含13种禁用农药),总检出率为98.5%,农药检出量为0.001~11.7 mg/kg。检出农药的慢性膳食摄入风险(%ADI)为0.003%~3.142%,急性膳食摄入风险(%ARfD)为0.022%~26.667%,风险均远低于100%,表明陈皮中农药的膳食暴露风险处于较低水平。6种有机磷农药的慢性累积暴露危害指数为0.942,略小于1,表明风险虽可控但需要关注。风险排序结果表明,陈皮中有16种为中高风险农药,应在生产和安全监管中重点关注。     

消化道肿瘤患者术前营养状况分析

为了解消化道肿瘤患者术前的营养状况,给制订术前患者营养干预方案提供依据,运用NRS-2002和24 h回顾膳食调查法对120例消化道肿瘤患者术前进行了营养评价和膳食调查.结果表明,NRS-2002评分结果,有67.5%患者存在营养不良风险,且年龄越大营养不良风险发生率越高;膳食调查结果,消化道肿瘤患者术前实际营养素摄入...     

Quantification of urinary N-acetyl-S- (propionamide)cysteine using an on-line clean-up system coupled with liquid chromatography/tandem mass spectrometry

Acrylamide has been reported to be present in high-temperature processed foods and normal processed food intake could lead to significant acrylamide exposure. Acrylamide in vivo can be conjugated with glutathione in the presence of glutathione transferase. This conjugation product is further metabolized and excreted as N-acetyl-S-(propionamide)cysteine (NASPC) in the urine. NASPC could be considered a biomarker for acrylamide exposure. The objective of this study was to develop a highly specific, rapid and sensitive method to quantify urinary NASPC, serving as a biomarker for acrylamide exposure assessment. Isotope-labeled [13C3]NASPC was successfully synthesized and used as an internal standard. This urine mixture was directly analyzed using a newly developed liquid chromatographic/tandem mass spectrometric method coupled with an on-line clean-up system. The detection limit for this method was estimated as < 5 microg l(-1)(0.4 pmol) on-column. The method was applied to measure the urinary level of NASPC in 70 apparently health subjects. The results showed that the NASPC urinary level was highly associated with smoking. Smokers had a significantly higher urinary NASPC level (135 +/- 88 microg g(-1) creatinine) than non-smokers (76 +/- 30 microg g(-1) creatinine). A highly sensitive and selective LC/MS/MS isotope dilution method was successfully established. With an on-line clean-up system, this system is capable of routine high-throughput analysis and accurate quantitation of NASPC in urine. This could be a useful tool for health surveillance for acrylamide exposure in a population for future study.     

Determination of sugars by liquid chromatography with post-column catalytic derivatization and fluorescence detection

Summary A method for the determination of reducing sugars such as fructose and glucose and nonreducing sugar such as sucrose by high performance liquid chromatography followed by an acidic hydrolysis and a derivatization with benzamidine has been developed. After separation of sugars on a gel column packed with a polymer-based cation exchange material (Sugar-Pak I, Waters-Millipore), the sucrose is first hydrolysed in a solid phase reactor to convert it into reducing subunits. A post-column fluorigenic reaction with benzamidine under alkaline condition allows the selective determination of both natural and converted reducing carbohydrates.This procedure has proven to be selective (fluorigenic detection) and highly sensitive (allowing detection as little as picomoles amounts), reproducible and linear over a broad range of concentrations: 5×10^–4 to 1.0×10^–2 M.The applicability of this method to natural matrices such as plant extracts and beverages is also described. The sugar content of a barley extract has been determined and compared with a specific enzymatic test. The determined sugar content of natural and commercial lemon juices as well as of Cola beverages has been compared with those found by the conventional LC refractive index analytical procedure. In all cases, the results were comparable and were within the experimental errors of the methods.     

Aluminium in tea—concentrations, speciation and bioavailability

Tea (Camellia sinensis) is one of a few plants accumulating aluminium (Al), making tea a major source of dietary Al intake. This paper reviews published studies on the concentrations, speciation and bioavailability of Al in tea. With very few exceptions, the total concentration of Al in tea infusions is in the range 1-6 mg l⁻¹. Probably more than 90% of this Al is bound to organic matter, but the nature of the organic species is unclear. Three studies using size exclusion chromatography provide evidence for Al species in the molecular mass (MM) range 4000-8500 Da, probably polyphenolic complexes. Two ultrafiltration studies indicate the presence of Al species with MMs above 10,000. The relative amount of the different organic Al species in tea infusions is unclear, and even the identity of any of these has not been demonstrated with certainty. A possible exception is Al trioxalate, which may be an important species based on evidence from two ²⁷Al-NMR studies. It seems fairly well established that drinking tea leads to measurable, but moderate increases in urinary Al excretion. However, the Al present in tea does not seem to be much more bioavailable than that from other dietary sources. Even so, it cannot be dismissed that tea infusions may contain particularly bioavailable and neurotoxic compounds such as Al maltolate, but this is at present speculative.     

Dissipation pattern and dietary risk assessment of some commonly used insecticides on tomato (Solanum lycopersicum L.)

Pesticide residue in food commodities is a serious concern in relation to consumer safety and also the most significant barrier in the trade of food commodities. The dissipation pattern of four insecticides, namely novaluron, λ-cyhalothrin, imidacloprid, and fenazaquin, was evaluated on tomato fruits and cropped soil. The residues were extracted using the QuEChERS analytical method and quantized using a gas chromatograph with electron capture detector, gas chromatograph mass spectrometer, and high-performance liquid chromatography with photo diode array detector. The analytical method was validated using parameters like recovery, linearity, accuracy, matrix effect, and specificity, with limit of detection and limit of quantitation established to be 0.01 and 0.05 mg/kg, respectively, for all the pesticides. The average initial deposits (samples collected after 2 h of application) at the recommended dose of novaluron, λ-cyhalothrin, imidacloprid, and fenazaquin were 0.593, 0.293, 0.227, and 0.431 mg/kg on tomato fruits, respectively, and were below the limit of quantification in soil. The pre-harvest interval of 17, 8, 1, and 13 days was suggested for novaluron, λ-cyhalothrin, imidacloprid, and fenazaquin on tomato, respectively. Risk assessment studies revealed that all pesticides under study are safe and do not pose any threat to humans as theoretical maximum dietary intake is less than the maximum permissible intake and acceptable daily intake.     

Quality assessment of a formulated Chinese herbal decoction, Kaixinsan, by using rapid resolution liquid chromatography coupled with mass spectrometry: A chemical evaluation of different historical formulae

Kaixinsan is an ancient Chinese herbal decoction mainly prescribed for patients suffering from mental depression. This decoction was created by Sun Si-miao of Tang Dynasty (A.D. 600) in ancient China, and was composed of four herbs: Radix and Rhizome Ginseng, Radix Polygalae, Rhizoma Acori Tatarinowii and Poria. Historically, this decoction has three different formulations, each recorded at a different point in time. In this study, the chemical compositions of all three Kaixinsan formulae were analyzed. By using rapid resolution LC coupled with a diode-array detector and an ESI triple quadrupole tandem MS (QQQ-MS/MS), the Radix and Rhizome Ginseng-derived ginsenosides including Rb(1), Rd, Re, Rg(1), the Radix Polygalae-derived 3,6'-disinapoyl sucrose, the Rhizoma Acori Tatarinowii-derived α- and β-asarone and the Poria-derived pachymic acid were compared among the three different formulations. The results showed variations in the solubility of different chemicals between one formula and the others. This systematic method developed could be used for the quality assessment of this herbal decoction.     

Chloride attachment negative-ion mass spectra of sugars by combined liquid chromatography and atmospheric pressure chemical ionization mass spectrometry

A method has been developed for the rapid determination of sugars, including molecular weight measurements, using high-performance liquid chromatography coupled with negative-ion, atmospheric-pressure chemical-ionization mass spectrometry. The chromatography was carried out on a 250 x 4 mm I.D. column packed with 7 microns NH2-silica. The mobile phase consisted of a high percentage of methanol or acetonitrile with a small amount of chloroform. During the mass spectrometry, a strong base is formed from the polar solvent molecules by corona discharge, followed by ion-molecule reactions in the chemical ionization ion source (e.g. the methoxy anion is formed from methanol). This strong base reacts with the chloroform, generating chloride ions, which in turn react with the neutral sugar molecules as they elute from the chromatograph. The chloride ion and sugar interactions yield chloride-attachment ions, which are further stabilized by successive collisions. In this method, authentic monosaccharides and some oligosaccharides show dominant quasi-molecular ions, [M + Cl]-, with little fragmentation, and its particularly useful for the molecular weight determination of sugars.     

Analysis of biomass sugars using a novel HPLC method

The precise quantitative analysis of biomass sugars is a very important step in the conversion of biomass feedstocks to fuels and chemicals. However, the most accurate method of biomass sugar analysis is based on the gas chromatography analysis of derivatized sugars either as alditol acetates or trimethylsilanes. The derivatization method is time consuming but the alternative high-performance liquid chromatography (HPLC) method cannot resolve most sugars found in biomass hydrolysates. We have demonstrated for the first time that by careful manipulation of the HPLC mobile phase, biomass monomeric sugars (arabinose, xylose, fructose, glucose, mannose, and galactose) can be analyzed quantitatively and there is excellent baseline resolution of all the sugars. This method was demonstrated for standard sugars, pretreated corn stover liquid and solid fractions. Our method can also be used to analyze dimeric sugars (cellobiose and sucrose).     

Analysis of formose sugar and formaldehyde by high-performance liquid chromatography

Formose sugar and formaldehyde (HCHO) were analyzed using high-performance liquid chromatography (HPLC) utilizing a CarboPac PA1 column (Dionex) and pulsed amperometric detection. This HPLC system was unsuitable for the analysis of formose sugar and HCHO and thus reducing sugars and unconverted HCHO were determined by endowing them with charges through a derivatization method using 2,4-dinitrophenylhydrazine. The separation and detection of compounds were performed by three Chromolith RP-C18 columns (Merck) and diode array detection, at a wavelength of 360 nm ultraviolet light, respectively. Lower sugars (except HCHO) showed some instabilities when the derivatized samples were kept for the extended periods of time. For C₅ and consecutive higher sugars, a certain derivatization time was necessary. In the present case (formose reaction with partial HCHO conversion), approximately 18 h may be a reasonable compromise for the derivatization reaction. A derivatization agent to compound mole ratio of up to 100:1 was required to complete the derivatization of C₄ and higher sugars. However, the analysis of C₄ and consecutive higher sugars is problematic for example due to overlapping of peaks or branched-chain sugars.