Determination of nitrite in cured meats by ion-exclusion chromatography with electrochemical detection

A rapid liquid chromatographic (LC) method was developed for a sensitive determination of nitrite in cured meats, using ion-exclusion chromatographic separation and electrochemical detection (IEC-EC). The current AOAC colorimetric method requires 2 h shaking in a steam bath to eliminate interference from reducing compounds such as ascorbic acid. In the present method, nitrite was analyzed in the presence of ascorbic acid without interference, and the extraction time was reduced to 1 min. The extracted nitrite was determined by ion chromatography using anion-exclusion/HS column and amperometric detector equipped with platinum or glassy carbon electrode operating at +1.0 V vs Ag/AgCl reference electrode. The detection limit was 1 ppb as NO2-. The recoveries of 50 ppm nitrite added to frankfurter and meat stick were 103 and 99.6%, respectively, with relative standard deviations less than 4%. The high speed, sensitivity, and selectivity make the new method a useful alternative to the AOAC colorimetric method.     

Chemiluminescence analysis of nitric oxide in small-volume samples by a modified injection method

To detect NO in samples of the headspace of static cores, a sample-mixing unit was developed which made it possible to detect NO in small-volume samples (50l to 10 ml) by a continuous flow chemiluminescence NO analyzer. The sample-mixing unit enables the detector of the NO analyzer to reach a steady-state response. The detection limit was 3.5 pmol NO in a sample, which is equivalent to 8.5 ppbv NO in a 10-ml sample or 85 ppbv No in a 1-ml sample at room temperature. The coefficient of variation between replicates varied between 6.4% at low NO levels and 0.6% at high NO levels. The linear response was good, theR ² value was above 0.99 even in the lowest detection range. The sample-mixing unit in combination with an NO analyzer provides a sensitive, reliable, easy to use method for the detection of NO in incubation systems with small headspace volumes such as static cores.     

Determination of monounsaturated alkyl side chain 2-alkylcyclobutanones in irradiated foods

The 2-alkylcyclobutanones (2-ACBs) are formed from triglycerides by irradiation treatment and may be used as markers for this type of food processing. This paper describes a detection method for the analysis of monounsaturated alkyl side chain 2-ACBs, which is formed upon irradiation from monounsaturated fatty acids which frequently are the most abundant fatty acids in foods. The estimated radioproduction yields of the cis-2-(dodec-5'-enyl)-cyclobutanones (cis-2-dDeCB) and the cis-2-(tetradec-5'-enyl)-cyclobutanones (cis-2-tDeCB) were 1.0 +/- 0.5 and 0.9 +/- 0.2 nmol.mmol(-1) precursor fatty acid.kGy(-1), respectively, being similar to that of saturated 2-ACBs. The stability study of the s- and mu-2-ACBs in poultry meat samples irradiated at 10 kGy and stored for 3-4 weeks at 4 degrees C and 25 degrees C showed that these compounds undergo some transformation, their amounts being reduced by about 50%. These storage losses did not depend on the saturation state of the alkyl side chain. The EI-MS detection limit of 2-tDeCB is 3 times higher (0.6 pmol) than that of 2-dodecylcyclobutanone (0.2 pmol). Consequently, when the oleic acid content of the analyzed food exceeds the content of palmitic acid by a factor of 3, it would be of an advantage to apply 2-tDeCB as a marker for detection of the irradiation treatment.     

Determination of ascorbigens in autolysates of various Brassica species using supercritical fluid chromatography

A new method of analysis based on normal phase supercritical fluid chromatography (SFC) has been developed for investigation of ascorbigens [2-C-(indol-3-ylmethyl)-beta-L-xylo-3-hexulofuranosonic acid gamma-lactone derivatives]. This method has been adapted to preparative isolation and quantitative determinations of individual ascorbigens comprising ascorbigen, neoascorbigen, and 4-methoxyascorbigen. The structures of these compounds have been revealed from 1D ((1)H, (13)C) and 2D (COSY, HMQC, HMBC) NMR experiments. The developed SFC method had an acceptable linearity for the ascorbigens with correlation coefficients (R(2)) > 0.9995 (n = 10) in the range of 0.13-4.9 nmol injected, detection limits were below 13 pmol, retention time stabilities were excellent, and relative response factors have been determined. The SFC method has been used for determination of ascorbigens produced during autolysis of indol-3-ylmethylglucosinolates in various Brassica vegetables and rapeseed seedlings. Generally, 30-60% of the indol-3-ylmethylglucosinolates in the plants were transformed into ascorbigens, with the concentration in autolysates varying from 0.51 +/- 0.002 to 3.72 +/- 0.21 micromol/g of dry weight (DW) for ascorbigen, from 0.05 +/- 0.01 to 2.42 +/- 0.23 micromol/g of DW for neoascorbigen, and from 0.03 +/- 0.002 to 0.84 +/- 0.07 micromol/g of DW for 4-methoxyascorbigen.     

Analysis of nitrate ion in nettle (Urtica dioica L.) by ion-pair chromatographic method on a C30 stationary phase

Nitrate ion is a frequent pollutant not only in soil and natural water resources but in vegetables and foods as well. In our study we focused on nettle due to its increased ability to accumulate nitrate ions. A new, simple method for the separation and determination of nitrate ion based on reversed-phase ion-pair chromatography has been elaborated. A new four-step sample pretreatment method enables the precipitation of proteins and oxidative degradation of compounds that may disturb the identification of the nitrate ion: (1) extraction of the total nitrate content, (2) precipitation of proteins with acetonitrile, (3) oxidative degradation of the organic contaminants with H2O2, (4) evaporation of the solvent and taking up of the residue in water. The chromatographic separations were carried out on a high-density C30 stationary phase under isocratic conditions. The optimal mobile-phase composition was 10% (v/v) acetonitrile and 90% (v/v) 20 mmol L(-1) phosphate buffer, containing 2 mmol of tetrabutylammonium hydroxide at pH 6.0. The method could also be used for the separation of IO3(-), SeO3(2-), BrO3(-), NO2(-), Br-, SeO4(2-), and I- ions. The validated method is sensitive (the detection limit is 0.18 ng of nitrate ion). The method is linear in a high concentration range (0.031-30.66 microg mL(-1)). Recoveries varied between 98% and 103%. Reproducibility of the elaborated sample pretreatment method showed 1.54%. The method can be used for the determination of nitrate ion from different plants.     

液质联用法同时测定猪粪便中16种（氟）喹诺酮类药物残留

为检测猪粪便中氟喹诺酮类药物残留,建立了高效液相色谱-串联质谱方法,可同时测定猪粪便中环丙沙星、诺氟沙星等16种氟喹诺酮类药物的残留量。样品经50%硝酸镁-4%氨水混合溶液提取,过HLB固相萃取小柱净化,HPLC-MS/MS多反应监测模式下进行定性及定量分析,16种氟喹诺酮类药物在40～200ng·mL^-1浓度范围内呈现良好线性关系,西诺沙星和沙拉沙星检出限为0.005μg·g-1,其他FQs药物检出限均为0.002μg·g-1。添加浓度为0.05、0.5、1.0μg·g-1时,方法平均回收率为51.4%～109.3%,RSD小于20%。该方法前处理简单、快速、灵敏、准确,仅使用少量有机试剂,检测成本低,对环境危害小。     

三氯乙酸提取硅作为水稻缺硅诊断指标的可行性

Potassium chloride, Na-pyrophosphate,CuCl2,NH4-oxalate,dithionit-citrate-bicarbonate(DCB) and Na-citrate solutions were employed to etract aluminum(Al) and iron(Fe) sequentially and separately from 15 acidic soils located at the Mangshan Mountains,Hunan Province,China,Many evidences showed that separate pyrophosphate extracted mainly KCl-extractable Al,organo-Al complexes and some inorganic Al compounds,whereas separate CuCl2 extracted KCl-extractable Al and some organo-Al complexes,CuCl2 extracted much less amounts of Al than pyrophosphate did from the soils .Separate oxalate did not extract all KCl-pyrophosphate-CuCl2-oxalate seuentially extractable Al and Fe ,Also,separate DCB did not extract all KCl-pyrophosphate-CuCl2-oxalate-DCB sequentially extractable Al. The forms of Al extacted by oxalate and DCB from the soils were majorly noncrystalline.The interlayered materials of 1.4-nm intergrade minerals of the soils were attributed mainly to hydroxy Al polymers.     

New isoflavonoid glycosides and related constituents from astragali radix ( Astragalus membranaceus ) and their inhibitory activity on nitric oxide production

Twenty-four secondary metabolites, including 16 isoflavonoids, 7 astragalasides, and 1 benzoquinone, have been isolated from the roots of Astragalus membranaceus (Astragali radix). Among these isolated isoflavonoids, (-)-methylinissolin 3-O-β-d-(6'-acetyl)-glucoside (1), (-)-methylinissolin 3-O-β-d-{6'-[(E)-but-2-enoyl]}-glucoside (2), and calycosin 7-O-β-d-(6'-acetyl)-glucoside (3) have been identified as new compounds on the basis of spectroscopic analysis; (-)-methylinissolin 3-O-β-d-glucoside (4) was isolated from the natural products for the first time. The nitric oxide (NO) production inhibitory activity of the major compounds has been assessed in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. To identify A. membranaceus, a fingerprint method was developed by using a high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) method. Furthermore, characteristic peaks for the 11 major compounds in the chromatogram were unambiguously confirmed.     

树木年轮及其根部土壤元素含量的对数线性相关模型的验证及应用

Nine Platanus acerifolia(Ait),Willd.trees growing in the Nanjing Children Teacher‘s School,Nanjing,China ,were selected to determine the contents of 13 chemical elements both in the 1994‘s growth rings,C(Z,t),and in the soils near the roots,C′(Z,t),of the trees ,The results showed that the relationship be-tween C(Z,t) and C′(Z,t) followed the logarithmic liner correlation moedl.lgC′(Z,t),=a(Z) b(Z)lgC(Z,t),Based on this model the chrono-sequences of chemical element contents in the soils were reproduced fromt those ih the tree rings;i.e., the dynamic variations in the chemical element contents of the soils at the sites were trced.In this study the chrono-sequences of the chemical elements including Cd,Pb,Mn,Co and Zn in the soils near the roots of a Platanus acerifolia tree from 1957 to 1994 were established ,and the background values of Cd,Pb,Mn,Co and Zn in the soil were calcuated by taking the lowest values of the chrono-sequences of the element conents as upper limits of the background values.     

Determination of capsaicin and dihydrocapsaicin in capsicum fruits by liquid chromatography-electrospray/time-of-flight mass spectrometry

A simple, highly selective, sensitive, and reproducible liquid chromatography-electrospray ionization/time-of-flight mass spectrometry method has been developed for the direct and simultaneous determination of capsaicin and dihydrocapsaicin in Capsicum fruit extracts. Capsaicin and dihydrocapsaicin are the two major members of the so-called capsaicinoid family, which includes other minor analogues, and usually account for at least 90% of the pungency trait in Capsicum fruits. Chromatographic separation of capsaicin and dihydrocapsaicin was achieved with a reversed-phase chromatography column, using a gradient of methanol and water. Quantification was done using as an internal standard (4,5-dimethoxybenzyl)-4-methyloctamide, a synthetic capsaicin analogue not found in nature. Analytes were base-peak resolved in less than 16 min, and limits of detection were 20 pmol for capsaicin and 4 pmol for dihydrocapsaicin. The intraday repeatability values were lower than 0.5 and 12% for retention time and peak area, respectively, whereas the interday repeatability values were lower than 0.6 and 14% for retention time and peak area, respectively. Analyte recoveries found were 86 and 93% for capsaicin and dihydrocapsaicin, respectively. The method developed has been applied to the identification and quantification of capsaicin and dihydrocapsaicin in fruit extracts from different Capsicum genotypes, and concentrations found ranged from 2 to 6639 mg kg(-1).     

First derivative spectrophotometric determination of anhydrotetracyclines in tetracyclines

A rapid method is presented for detection and determination of anhydrotetracycline-HCl (ATC-HCl) and 4-epianhydrotetracycline-HCl (4-EATC-HCl) in tetracycline-HCl (TC-HCl). The method determines the 2 compounds as a sum, not individually. The first derivative absorption curve has a trough (D1) at 460 nm which is linearly related to concentration (1-10 mg/mL). ATC-HCl + 4-EATC-HCl content was determined in TC-HCl powder and capsules by the D1 and the compensation D1 spectrophotometric methods. The results were compared with those obtained using U.S. Pharmacopeia and British Pharmacopoeia methods.     

Quantitative determination of sulfur-containing wine odorants at sub parts per billion levels. 2. Development and application of a stable isotope dilution assay

[(2)H(10)]-4-Mercapto-4-methylpentan-2-one (d(10)-1), [(2)H(2)]-3-mercaptohexan-1-ol (d(2)-2), and [(2)H(5)]-3-mercaptohex-1-yl acetate (d(5)-3), deuterated analogues of impact odorants of wines, were used to determine quantitatively the natural compounds in white wines (Muscadet, Sauvignon, and Bacchus) with a stable isotope dilution assay using gas chromatography coupled either with ion trap tandem mass spectrometry (GC-ITMS-MS) or with atomic emission detection monitored on sulfur-selective acquisition (GC-AED). The thiol compounds were recovered from wines by liquid-liquid extraction, then purified from the wine extracts by covalent chromatography, and analyzed. The quantitative determination of 4-mercapto-4-methylpentan-2-one 1 in the wines that were analyzed was performed better with GC-AED than with GC-ITMS-MS under the conditions that were used. However, the detection limit of the method was higher than the odor threshold of 4-mercapto-4-methylpentan-2-one 1 in wine (5 vs 0.8 ng/L). The levels of this compound in the Sauvignon and Bacchus wines were much higher than its odor threshold, but it was not detectable in the Muscadet wines. On the contrary, GC-ITMS-MS was much more sensitive than GC-AED for detection of 3-mercaptohexan-1-ol 2 and 3-mercaptohex-1-yl acetate 3, and the detection limits were much lower than their odor thresholds in wine. The former compound was detected in all of the Muscadet wines that were analyzed at levels always higher than its odor detection threshold, while the latter occurred at levels higher than its odor threshold in only one Muscadet wine.     

中国南方典型地区阔叶林大气氮沉降通量研究

A one-year study in a typical red soil region of southern China was conducted to determine atmospheric nitrogen （N） fluxes of typical N compounds （NH3, NH4-N, NO3-N, and NO2） and contribution of three sources （gas, rainwater, and particles） to N deposition. From July 2003 to June 2004, the total atmospheric N deposition was 70.7 kg N ha^-1, with dry deposition accounting for 75% of the total deposition. Dry NH3 deposition accounted for 73% of the dry deposition and 55% of the total deposition. Moreover, NO2 contributed 11% of the dry deposition and 8% of the total deposition. Reduced N compounds （NH4^＋ and NH3） were the predominate contributors, accounting for 66% of the total deposition. Therefore, atmospheric N deposition should be considered when soil acidification and critical loads of atmospheric deposition on soils are estimated.     

Determination of benzoxazinone derivatives in plants by combining pressurized liquid extraction-solid-phase extraction followed by liquid chromatography-electrospray mass spectrometry

A new analytical method based on the use of pressurized liquid extraction (PLE) followed by solid-phase extraction with LiChrolut RP C18 cartridges was evaluated for the sample preparation, extraction, and cleanup of eight naturally occurring benzoxazinone derivatives, 2-beta-D-glucopyranosyloxy-4-hydroxy-1,4-benzoxazin-3-one, 2-beta-D-glucopyranosyloxy-4-hydroxy-7-methoxy-1,4-benzoxazin-3-one, 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA), 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one, 2-hydroxy-1,4-benzoxazin-3-one, 2-hydroxy-7-methoxy-1,4-benzoxazin-3-one, benzoxazolin-2-one, and 6-methoxybenzoxazolin-2-one in plant samples. Afterward, liquid chromatography-electrospray mass spectrometry, using the selected ion monitoring mode and internal standard (2-MeO-DIBOA, indoxyl-beta-D-glucoside, and quercetin-3-O-rutinoside) quantification method was performed. This paper demonstrates the effectiveness of the PLE method, in conjunction with sensitive and specific mass spectrometric detection, for the quantitative recovery of compounds of the benzoxazinone class from plants. The recoveries of the analytes ranged from 66 to 110% with coefficients of variation ranging from 1 to 14%. This method gave detection limits between 1 and 27 microg/g. The method was applied to foliage and roots of three different wheat cultivars, and the analytes were detected in the range of 11-3261 microg/g of dry weight.     

Novel nitric oxide and superoxide generation inhibitors, persenone A and B, from avocado fruit

One known, (2R)-(12Z,15Z)-2-hydroxy-4-oxoheneicosa-12,15-dien+ ++-1-yl acetate (1), and two novel compounds, persenone A (2) and B (3), have been isolated from avocado fruit (Persea americana P. Mill), as inhibitors of superoxide (O(2)(-)) and nitric oxide (NO) generation in cell culture systems. They showed marked inhibitory activities toward NO generation induced by lipopolysaccharide in combination with interferon-gamma in mouse macrophage RAW 264.7 cells. Their inhibitory potencies of NO generation (1, IC(50) = 3.6; 2, IC(50) = 1.2; and 3, IC(50) = 3.5 microM) were comparable to or higher than that of a natural NO generation inhibitor, docosahexaenoic acid (DHA; IC(50) = 4.3 microM). Furthermore, compounds 1-3 and DHA markedly suppressed tumor promoter 12-O-tetradecanoylphorbol-13-acetate-induced O(2)(-) generation in differentiated human promyelocytic HL-60 cells (1, IC(50) = 33.7; 2, IC(50) = 1.4; 3, IC(50) = 1.8; and DHA, IC(50) = 10.3 microM). It is notable that they were found to be suppressors of both NO- and O(2)(-)-generating biochemical pathways but not to be radical scavengers. The results indicate that these compounds are unique antioxidants, preferentially suppressing radical generation, and thus may be promising as effective chemopreventive agent candidates in inflammation-associated carcinogenesis.     

Liquid chromatographic determination of fluridone aquatic herbicide and its metabolite in fish and crayfish

A residue method is described for determination of the aquatic herbicide fluridone (1-methyl-3-phenyl-5-[3-(trifluoromethyl)phenyl]-4(1H)-pyridinone) and its metabolite (1-methyl-3-(4-hydroxyphenyl)-5-[3-(trifluoromethyl) phenyl]-4(1H)-pyridinone) in fish and crayfish tissues. Both compounds are extracted from tissues with methanol, and the extracts are subjected to acidic hydrolysis to release conjugated forms of fluridone and the metabolite. Sample extracts are purified by liquid-liquid partitioning and Florisil Sep-Pak column chromatography. Both compounds are separated and measured by reverse phase liquid chromatography with UV detection at 313 nm. In the absence of interfering peaks, the method has a detection limit of approximately 0.04 ppm of either compound. Overall, recoveries averaged 96% for fluridone and 78% for the metabolite for all tissue types combined.     

Development of a solid-phase cleanup and portable rapid flow-through enzyme immunoassay for the detection of ochratoxin a in roasted coffee

A membrane-based flow-through enzyme immunoassay (patent application pending) for the detection of ochratoxin A (OA) in roasted coffee was developed. First, an extraction and solid-phase cleanup method was developed. A high partition coefficient for OA in the mobile phase was achieved by using methanol/5% aqueous NaHCO(3) as the sample extraction and cleanup solvent. The solid-phase (aminopropyl) cleanup was developed to chromatographically elute OA but retain cross-reacting compounds. Without using aminopropyl cleanup, cross-reacting compounds resulted in 100% false positives for both flow-through enzyme immunoassay and HPLC methods. However, after cleanup with aminopropyl, no false positives were observed. The flow-through results were visually evaluated. The sensitivity achieved for the flow-through was 4 microg kg(-1) in spiked roasted coffee. The assay was used to screen roasted coffee samples. Results were confirmed with HPLC with a detection limit of 1 microg kg(-1).     

Development of a new high-performance liquid chromatography-electrospray ionization time-of-flight mass spectrometry method for the determination of low molecular mass organic acids in plant tissue extracts

A liquid chromatography-electrospray ionization time-of-flight mass spectrometry method has been developed for the direct and simultaneous determination of 10 low molecular mass organic acids in different plant tissue extracts. The method does not include a derivatization step. Quantification was accomplished using (13)C-labeled malic and succinic acids as internal standards. Good limits of detection (0.05-1.27 pmol) were obtained for malic, 2-oxoglutaric, succinic, quinic, shikimic, cis-aconitic, and citric acids, whereas for oxalic, ascorbic, and fumaric acids limits of detection were 255, 25, and 11 pmol, respectively. Repeatability values for the retention time and peak area were <5%, with the exception of ascorbic acid. Analyte recovery was between 92% and 110% in most cases, with the exception of oxalic (39-108%), 2-oxoglutaric (44-69%), and ascorbic (22-86%) acids, which may require specific extraction procedures and use of the corresponding (13)C-labeled organic acid as internal standards to improve accuracy. The method was applied to three types of plant materials: sugar beet leaf extracts, tomato xylem sap, and commercial orange juice.     

Anti-inflammatory effects of phenolic compounds isolated from the fruits of Artocarpus heterophyllus

Artocarpus heterophyllus Lam is a large evergreen tree cultivated throughout Southeast Asia for its fruits. Its leaves and roots have been used for medicinal purposes. The aim of this work was to study the in vitro anti-inflammatory effects of phenolic compounds isolated from the ethyl acetate extracts of the fruits of Artocarpus heterophyllus. Three phenolic compounds were characterized as artocarpesin [5,7,2',4'-tetrahydroxy-6-(3-methylbut-3-enyl) flavone] ( 1), norartocarpetin (5,7,2',4'-tetrahydroxyflavone) ( 2), and oxyresveratrol [ trans-2,4,3',5'-tetrahydroxystilbene] ( 3) by spectroscopic methods and through comparison with data reported in the literatures. The anti-inflammatory effects of the isolated compounds ( 1- 3) were evaluated by determining their inhibitory effects on the production of proinflammatory mediators in lipopolysaccharide (LPS)-activated RAW 264.7 murine macrophage cells. These three compounds exhibited potent anti-inflammatory activity. The results indicated that artocarpesin ( 1) suppressed the LPS-induced production of nitric oxide (NO) and prostaglandin E 2 (PGE 2) through the down-regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) protein expressions. Thus, artocarpesin ( 1) may provide a potential therapeutic approach for inflammation-associated disorders.     

Antioxidant properties of sour cherries (Prunus cerasus L.): role of colorless phytochemicals from the methanolic extract of ripe fruits

Many edible plant metabolites are known to be useful as cellular antioxidants. In the search for antioxidative chemicals from native fruits of the Campania region of Italy, Prunus cerasus L., an acidic cherry widely used for culinary purposes, has been studied. Fruit crude extracts (MeOH, EtOAc, and hexane) were submitted to an antioxidative screening using specific assay media characterized from the presence of highly reactive radical species (DPPH*, ABTS*+, O2*-, NO) or lipoperoxidation markers. The reducing power of the samples was also determined. It was observed that the most polar extracts in MeOH and EtOAc were able to exercise a massive and dose-increasing antioxidative capacity. The peculiar efficacy of the same extracts was revealed by investigating their protein and deoxyribose oxidation capacity. A preliminary analysis of total phenol, flavonoid, and anthocyanin contents together with biological screening data put the basis on P. cerasus fruit phytochemical investigation of methanolic extract. Twenty secondary metabolites were isolated and characterized by spectroscopic (especially 1D and 2D NMR) and spectrometric techniques. 1-(4-Hydroxyphenyl)-1,2-ethanediol-1,2-bis-1-O-beta-D-glucopyranoside (3), (4-hydroxy-3-methoxyphenyl)methanol-1-O-beta-D-gentiobioside (4), epicatechin-3-malate (14), and epicatechin-3-(1'-methyl)malate (15) were isolated for the first time. All of the compounds were evaluated for their radical scavenging activity on DPPH*, O2*-, and NO. Flavonoids and quinic acid derivatives were found to be the more antioxidative substances.