Comparison of various extraction methods for policosanol from rice bran wax and establishment of chromatographic fingerprint of policosanol

A capillary gas chromatographic (GC) method has been developed for the separation and determination of policosanol components extracted from rice bran wax. A Varian CP-sil 8 CB column was employed, and an oven temperature was programmed. Gas chromatography-mass spectrometry (GC-MS) was used to identify the composition of policosanol. Quantitative analysis was carried out by means of hydrogen flame ionization detector (FID) with dinonyl phthalate (DNP) as internal standard. The results indicated that the extract obtained by dry saponification has the highest contents of octacosanol and triacontanol among extracts by all used extraction methods including dry saponification, saponification in alcohol, saponification in water (neutralized and non-neutralized), and transesterification. Meanwhile, the GC-MS fingerprint of policosanol extracted by dry saponification has been established. Euclidean distance similarity calculation showed remarkable consistency of compositions and contents among 12 batches of policosanol from a rice bran wax variety. This protocol provided a rapid and feasible method for quality control of policosanol products.     

Liquid chromatographic determination of desoxycorticosterone acetate in oil injections.

To determine desoxycorticosterone acetate in oil injections, reverse phase partition chromatography on silanized, purified siliceous earth was used to separate the corticosteroid ester from the bulk of the oil vehicle. The latter was retained on the column while the steroid and the sterol and triterpenoid fractions of the oil were eluted. An internal standard was added to this eluate, which was then subjected to reverse phase high performance liquid chromatography (HPLC). The desoxycorticosterone acetate was quantitatively separated in the HPLC procedure from any free desoxycorticosterone, preservatives, and minor components of the oil. The suitability of the HPLC procedure was verified with a number of C18 packing materials, both pellicular and microparticular. The desoxycorticosterone acetate was adequately resolved from the internal standard, progesterone, with most C18 packing materials evaluated. The proposed procedure provides a suitable stability-indicating assay for desoxycorticosterone acetate in oil injections.     

Gas chromatographic detection of the mycotoxin zearalenone in blood serum

A sensitive gas chromatographic method for the quantitative analysis of zearalenone in blood serum is described. Zearalenone is eluted from blood serum by column chromatography followed by base-acid extraction with dichloromethane as the organic phase. After epicoprostanol (internal standard) is added, the sample is evaporated to dryness, derivatized, and injected onto the gas chromatographic column. A number of silylating agents and reaction conditions were investigated. Derivatizing zearalenone with N-methyl-N-trimethylsilyltrifluoroacetamide in the presence of acetone at room temperature for at least 2 hr gave best results. Sensitivity limit is < 0.5 ng injected, equivalent to 100 ng zearalenone/mL blood serum. A linear standard curve is observed when 0.5-30 ng zearalenone derivative is injected onto the Perkin-Elmer gas chromatograph. For quantitation, a standard curve is prepared by plotting amounts of zearalenone (ng) injected vs. ratios for peak areas of zearalenone and epicoprostanol derivatives. The internal standard procedure improves the precision by minimizing variations in sample injections and detector response. Percent recovery from blood serum is 68-75 in the range of 1.6-8.0 micrograms zearalenone/mL blood.     

三峡水库消落带淹水—落干交替下紫色土力学特性变化模拟

三峡水库消落带在高压淹水浸泡和干湿交替作用下,土壤力学特性必然发生变化,影响土体抗侵蚀性能,破坏库岸稳定性。目前对消落带土壤力学特性变化及其对淹水—落干驱动的响应尚不明确。以三峡水库消落带广泛分布的紫色土为研究对象,模拟水压作用和淹没—出露交替过程,研究紫色土应力—应变和抗剪强度参数对高压淹水和干湿交替作用的响应。结果表明:水柱压力、干湿交替次数和干湿交替幅度都能显著驱动紫色土力学特性发生变化。与对照组相比,随着水柱压力增加,紫色土刚度逐渐增强,黏聚力降低幅度变小,内摩擦角增大。随干湿交替次数增加,紫色土刚度逐渐减小;内摩擦角有所增大,黏聚力的变化趋势与之相反,且随干湿交替次数的增加而显著降低。随干湿交替幅度增加,抗剪强度迅速降低(相同围压下),黏聚力增大,内摩擦角降低;与对照组相比,干湿交替幅度越小,土壤的内摩擦角增大越多,黏聚力降低越多。     

Liquid chromatographic method for quantitative determination of free fatty acids in butter

A liquid chromatographic method has been developed for the determination of free fatty acids in butter. The fatty acids are converted to the p-bromophenacyl esters, via a crown ether-catalyzed reaction, without separation from the other butter components. The esters are separated on a C18-bonded silica column by using an acetonitrile-water solvent gradient and quantitated using the ester of heptadecanoic acid as internal standard. C16 and C18:1 co-elute in the acetonitrile-water system but are separated using an isocratic methanol-acetonitrile-water system. Limits of detection range from 7 ng for butyric acid to 45 ng for linoleic acid. The average coefficient of variation (n = 10) for 10 free fatty acids from a butter was 5.83%.     

Effects of addition of clay minerals on the fatty acid fraction of a podzol soil

The Al horizon of a sandy podzol soil, with a very small proportion of clay-sized material, was sampled and homogenized. Samples, both with and without supplementation with 150 g clay minerals kg^?1, natural heteroionic bentonite (saturated with Na, Ca, Mg, K), Mg-bentonite or Fe-kaolinite, were moistened to two-thirds of water-holding capacity and incubated at 28°C for 16 weeks. After incubation, lipids were extracted with 3:1 light petroleum: ethylacetate mixture in a Soxhlet apparatus as unbound components (directly extractable by organic solvents) and bound components (only extractable after treatment with acids). Monoacids (free and esterified), diacids, ketoacids and hydroxyacids were isolated from the two lipidic fractions (unbound and bound) by column chromatography. The different components of each family were analyzed by gas chromatography (GC) or gas chromatography-mass spectrometry (GC-MS). Incubation without supplementation generated a selective decrease in abundance of free mono-acids in unbound and bound fractions. The decrease was in inverse proportion to the chain-length of components. Soil supplementation with bentonite (homoionic or heteroionic) did not induce an overall quantitative change, but promoted a transfer of fatty acids from the unbound to the bound fraction, increased the exchange between free and esterified monoacids (transesterification) and stimulated hydrolysis of glycerides. In contrast, soil supplementation with Fe-kaolinite caused much decomposition or biotransformation of monoacids but it did not induce change either in the intensity of transesterification in the wax ester fraction or in the intensity of hydrolysis in the glyceride fraction.     

Analysis of pharmaceutical dosage forms for oxfendazole: II. Simultaneous liquid chromatographic determination of oxfendazole and trichlorfon in equine paste

A reverse phase liquid chromatographic procedure is described for the simultaneous determination of oxfendazole [2-(methoxycarbonylamino)-5-phenylsulfinylbenzimidazole] and trichlorfon [(2,2,2-trichloro-1-hydroxyethyl)phosphonic acid dimethyl ester] in equine paste. The sample is extracted by sonication in methanol. Insoluble excipients are removed by centrifugation and an aliquot plus internal standard are diluted with dilution solvent (water-acetonitrile-phosphoric acid, 80 + 20 + 1). The samples are filtered and injected onto a Partisil-5 ODS-3 column with acetonitrile-0.01 M phosphate buffer pH 6.0 (20 + 80) as mobile phase. Method specificity is confirmed using an absorbance rationing technique. The method yields mean recoveries of 100.9 and 100.0% for trichlorfon and oxfendazole, respectively. Dependence of chromatographic performance characteristics on mobile phase organic content, pH, and buffer concentration is also reported.     

Steviol quantification at the picomole level by high-performance liquid chromatography

A simple and highly sensitive reversed-phase high-performance liquid chromatographic method (RP-HPLC) has been developed for the determination of steviol (SV) using dihydroisosteviol (DHISV) as an internal standard (IS). SV and DHISV were derivatized by reaction of the acids with 4-(bromomethyl)-7-methoxycoumarin in an aprotic solvent (DMF or acetone). The resulting ester derivatives were separated on an ODS column (250 x 4.6 mm i.d., 5 microm particle size) using fluorescence detection with excitation at 321 nm and emission at 391 nm. The mobile phase consisted of acetonitrile/water (80:20 v/v) with a flow rate of 1 mL min(-)(1). A linear relationship was observed for concentrations between 0.5 and 50 microg/mL of SV, and the detection limit was 100 pg. For application of this method to samples of beer fortified with stevioside, a simple procedure for extraction of the beer with diethyl ether and derivatization in DMF was applied. Whereas beer samples spiked with SV gave a linear response over the range 0.1-15 microg/mL beer, no SV could be detected in beer samples enriched in stevioside that had been stored for over 3 years. The application of the method to plant samples involved preparation of an acid fraction containing the SV analyte, derivatization, and sample cleanup using small silica columns and thin-layer chromatography. A sensitive determination of 594 ng of steviol present in 100 mg of dry plant material was performed with high precision and accuracy.     

Parallel synthesis: a new approach for developing analytical internal standards. Application to the analysis of patulin by gas chromatography-mass spectrometry

The polymer-assisted reaction of 4-(hydroxymethyl)furan-2(5H)-one (4HM2F) with 21 carboxylic acids using polystyrene-carbodiimide (PS-carbodiimide) yielded an ester library. Four of the esters, (5-oxo-2,5-dihydrofuran-3-yl)methyl acetate (IS-1), (5-oxo-2,5-dihydrofuran-3-yl)methyl butyrate (IS-2), (5-oxo-2,5-dihydrofuran-3-yl)methyl 2-methylpropanoate (IS-3), and (5-oxo-2,5-dihydrofuran-3-yl)methyl chloroacetate (IS-4), were tested as internal standards for the quantification of patulin in apple juice by gas chromatography-mass spectrometry in the selected ion monitoring mode (GC-MS-SIM). The developed method combines an AOAC official extractive step and a GC-MS-SIM analysis. Using a chromatographic column containing trifluoropropylmethylpolysiloxane as the stationary phase and IS-1 as the internal standard, it was possible to perform an accurate and precise quantification of underivatizated patulin in apple juice at concentrations down to 6 microg/L. A detection limit of 1 microg/L was established.     

Gas chromatographic method for determination of fenitrothion in fenitrothion technical and in formulated products: collaborative study

A gas chromatographic (GC) method for determination of fenitrothion in fenitrothion technical and formulated products has been subjected to a collaborative study with 7 participating laboratories. Formulations are extracted with chloroform containing dibutyl sebacate as an internal standard and are analyzed by gas chromatography using an OV-210 column. Collaborators were furnished matched pairs of technical product and water-dispersible powder and emulsifiable concentrate formulations. Relative standard deviations for reproducibility (RSDR) for the paired samples were 0.54, 1.00, and 1.56%, respectively, for technical fenitrothion, water-dispersible powders, and emulsifiable concentrates. The method has been approved interim official first action as an alternative to the present official first action AOAC method 6.A19-6.A24, which uses a polyphenyl ether, 6 ring (PPE-6R) column packing and fluoranthene as internal standard.     

Gas-liquid chromatographic determination of capsaicin.

A simple and accurate method has been developed for the quantitative determination of capsaicin in capsicum spices and their oleosresins. Capsaicin is extracted with acetone and separated from interfering substances, using an activated alumina column. The column is first eluted with acetone, and then the capsaicin is eluted with acetone-methanol-water (75+25+2). The capsaicin is quantitated by gas-liquid chromatography, using a Carbowax 20M-Teflon column and piperine as an internal standard. Results can be calculated as per cent capsaicin or Scoville units. Recovery of added capsaicin averaged 101%.     

Gas chromatographic method for determination of fenitrothion in technical and in emulsifiable concentrate and water-dispersible powder formulations: collaborative study

A variety of column packings and internal standards were evaluated to determine the most satisfactory system to use in a gas chromatographic (GC) method for analysis of fenitrothion, technical and formulations. Fenitrothion and the most closely related isomer, O,O-dimethyl O-(4-methyl-3-nitrophenyl) phosphorothioate, were resolved on columns packed with OV-210 and with polyphenyl ether, 6-ring (PPE-6R). A method based on the separation of fenitrothion on a PPE-6R column with fluoranthene as internal standard was selected for use in a limited collaborative trial and later for use in a full-scale collaborative trial with 21 collaborators participating. Each collaborator was furnished matched pairs of samples of technical fenitrothion, emulsifiable concentrate, and water-dispersible powder. The coefficients of variation (CV) for the paired samples were 1.02, 1.11, and 1.01, respectively, for technical fenitrothion, emulsifiable concentrates, and water-dispersible powders. Data are also presented for an alternative method in which compounds are separated on an OV-210 column with dibutyl sebacate as the internal standard. The method has been adopted official first action.     

Simultaneous liquid chromatographic determination of creatinine and pseudouridine in bovine urine and the effect of sample pH on the analysis

A rapid, reliable method for the simultaneous determination of creatinine and pseudouridine is described. Both analytes were detected at an optimum wavelength of detection (262 nm), considering the relative levels present in bovine urine. Cimetidine was used as the internal standard and detected at its maximum wavelength of absorption (220 nm) on a separate channel. All three compounds were eluted within 15 min, using a 10 mmol/L phosphate buffer (pH 6.8)-methanol gradient on a C18 column. Creatinine data were found to be significantly dependent upon the pH of the sample. Recoveries of both analytes were above 96%. Lowest detectable levels of creatinine and pseudouridine were 0.28 nmol and 9.0 pmol, respectively. The use of internal standard resulted in a method with high precision (standard deviation of 1.42 mmol/L and 0.027 mmol/L for creatinine and pseudouridine), yet one that was simple and rapid.     

Determination of volatile N-nitrosamines in frankfurters containing minced fish and surimi

A method was developed to quantitatively measure volatile N-nitrosamines, particularly N-nitrosodimethylamine (NDMA), in cured minced fish or surimi-meat frankfurters. This method is free from artifact formation. First, 2 dry solid-phase extraction columns are prepared. Solvent is passed through the top column containing the fish-meat into a second column containing acid Celite. The eluate from the Celite column is then passed through a third column containing silica gel. Nitrosamines are eluted from the acid Celite column and then from the silica gel column into the same receiver. Recovery of the internal standard, N-nitrosoazetidine, added at the 10 ppb level, was 86.5%. In addition, a few samples of nitrite-treated salmon (lox) were also tested for N-nitrosamines. The results show that the method is applicable to samples containing nitrite-treated fish and fish-derived products.     

Determination of chlorophylls in Taraxacum formosanum by high-performance liquid chromatography-diode array detection-mass spectrometry and preparation by column chromatography

Taraxacum formosanum, a well-known Chinese herb shown to be protective against hepatic cancer as well as liver and lung damage, may be attributed to the presence of abundant carotenoids and chlorophylls. However, the variety and content of chlorophylls remain uncertain. The objectives of this study were to develop an high-performance liquid chromatography-diode array detection-mass spectrometry method for determination of chlorophylls in T. formosanum and preparation by column chromatography. An HyPURITY C18 column and a gradient mobile phase of water (A), methanol (B), acetonitrile (C), and acetone (D) could resolve 10 chlorophylls and an internal standard Fast Green FCF within 30 min with a flow rate at 1 mL/min and detection at 660 nm. Both chlorophylls a and a' were present in the largest amount (1389.6 μg/g), followed by chlorophylls b and b' (561.2 μg/g), pheophytins a and a' (31.7 μg/g), hydroxychlorophyll b (26.5 μg/g), hydroxychlorophylls a and a' (9.8 μg/g), and chlorophyllides a and a' (0.35 μg/g). A glass column containing 52 g of magnesium oxide-diatomaceous earth (1:3, w/w) could elute chlorophylls with 800 mL of acetone containing 50% ethanol at a flow rate of 10 mL/min. Some new chlorophyll derivatives including chlorophyllide b, pyropheophorbide b, hydroxypheophytin a, and hydroxypheophytin a' were generated during column chromatography but accompanied by a 63% loss in total chlorophylls. Thus, the possibility of chlorophyll fraction prepared from T. formosanum as a raw material for future production of functional food needs further investigation.     

Determination of benzene in polypropylene food-packaging materials and food-contact paraffin waxes

An analytical procedure was developed for determination of benzene in polypropylene food packaging and was adapted for determination of benzene in commercial paraffin waxes intended for food-contact use. The polymer was dissolved in hexadecane at 150 degrees C. The wax was melted in an 80 degrees C oven. A simple helium-sparging apparatus was used to remove the volatile chemical from the polymer or wax. The contaminant was collected in methanol, distilled water was added, and the resulting solution was analyzed by headspace gas chromatography. The instrument was equipped with a 30 m fused silica open tubular capillary column and a photoionization detector. Average recoveries of benzene from polymer and paraffin wax at low parts-per-billion concentrations were 63 and 70%, respectively. Limits of detection and quantitation for analysis of polypropylene were 8 and 17 ppb, respectively; the limit of quantitation for analysis of paraffin wax was 2 ppb. In several commercial polypropylene products examined, benzene levels ranged from none detected to 426 ppb. In 3 commercial waxes examined, concentrations of 16-73 ppb benzene were determined. The presence of benzene was confirmed by gas chromatography/mass spectrometry.     

Gas-liquid chromatographic determination of p-chloroacetanilide in acetaminophen

Gas-liquid chromatography GLC) coupled with column chromatography was used to accurately determine as little as 25 ppm p-chloroacetanilide in acetaminophen. p-Chloroacetanilide was eluted from a pH 8 phosphate-buffered diatomite partition column by using purified tetrachloroethylene (acetaminophen was retained). This solution was concentrated, internal standard (docosane) was added, and p-chloroacetanilide was determined by using a 0.9 m X 2 mm glass column packed with 3% Poly A 103 on Supelcoport and a flame ionization detector with electronic integration. Standard curves were linear for 10-100 ppm p-chloroacetanilide. Various chromatographic materials were investigated for optimal retention characteristics. High pressure liquid chromatography (HPLC) was also evaluated as an alternative; however, lack of reproducibility of the HPLC column favored the GLC procedure.     

Gas chromatographic-mass spectrometric determination of epoxidized soybean oil contamination of foods by migration from plastic packaging

A method for the quantitative determination of epoxidized soybean oil (ESBO) in foods is described. The procedure involves addition of a diepoxidized fatty acid ester internal standard, followed by lipid extraction from the food and transmethylation under basic conditions. Without further cleanup, the methylated fatty acid epoxides are derivatized to form 1,3-dioxolanes, which are then determined by capillary gas chromatography-mass spectrometry with selected ion monitoring. A detection limit of 2.0 mg/kg of epoxidized soybean oil in foods and a relative standard deviation of 7% have been achieved routinely. The method has been applied successfully to the analysis of cheeses, sandwiches, cakes, and microwave-cooked meals which have been contaminated with ESBO by migration from PVC film.     

Liquid chromatographic determination of taurine in vitamin premix formulations

A liquid chromatographic (LC) method is described for the determination of taurine in vitamin and vitamin-mineral premix formulations. The method involves extraction of taurine with 0.1 M bicarbonate buffer, followed by precolumn derivatization with dansyl chloride and LC using fluorescence detection. 6-Aminocaproic acid is used as an internal standard. A reverse phase analytical column and a mobile phase of 0.1 M acetate buffer solution (pH 7.2)-acetonitrile (75 + 25) are used. Vitamins, minerals, and other excipients in the premix formulations do not interfere in the determination. The method is simple, precise, and accurate.     

土壤中硝化抑制剂DMPP含量的气相色谱测定法

建立了一种检测土壤中吡唑类硝化抑制剂DMPP含量的气相色谱法。DMPP在NaOH溶液中可定量转化为DMP,从5种有机溶剂中选择氯仿作为DMP的萃取剂。本文采用吡啶做为内标物,内标校正因子fDMP和fDMPP分别为1.6775和3.3884。通过DB-1701气相色谱柱,氢火焰离子化检测器(FID)定量检测氯仿溶液萃取相中的DMP。本法添加回收率为97.4%～98.8%;RSD为0.70%～2.09%;土壤中DMPP的检出限为0.4mgkg-1。应用本法测定在潮棕壤中10℃、20℃和30℃时DMPP的降解半衰期分别为大于28d、14d和7d。