Rapid separation of gangliosides using strong anion exchanger cartridges

A method utilizing strong anion exchanger cartridges (InertSep SAX) was developed to separate gangliosides. Total lipids extracted from rat brain is able to be rapidly separated into neutral and acidic lipids rapidly. Neutral lipids were passed through the SAX cartridge while acidic lipids adsorbed onto the cartridge and were eluted by chloroform/methanol/4.0 M aqueous ammonium acetate (5:10:1, by volume). Moreover, various kinds of gangliosides (GM1, GD1a, GD1b, GT1b) were separated individually according to their characteristics by elution with increasing concentration of ammnonium acetate (0 - 4.0 M). The gangliosides yield of this procedure was higher than 95%.     

Rapid separation of neutral lipids,free fatty acids and polar lipids using prepacked silica sep-Pak columns

A method is described for the separation of neutral lipid, free fatty acid and polar lipid classes using small (600 mg), prepacked silica Sep-Pak columns. Combinations of hexane and methyltertiarybutylether were used to progressively elute cholesteryl ester first then triglyceride from the column. After column acidification, fatty acids were eluted followed by cholesterol. Recoveries of these lipids were 96% or greater. Polar lipids were eluted from the column using combinations of methyltertiarybutylether, methanol and ammonium acetate. Phospholipid classes could not be separated completely from each other. Phosphatidylethanolamine and phosphatidylinositol eluted together, whereas the more polar phosphatidylcholine, sphingomyelin and lysophosphatidylcholine were eluted as a second fraction. Recoveries of each phospholipid was greater than 98%.     

Two dimensional thin layer chromatographic separation of polar lipids and determination of phospholipids by phosphorus analysis of spots

Separation of polar lipids by two-dimensional thin layer chromatography providing resolution of all the lipid classes commonly encountered in animal cells and a sensitive, rapid, reproducible procedure for determination of phospholipids by phosphorus analysis of spots are described. Values obtained for brain and mitochondrial inner membrane phospholipids are presented.     

Complete separation of phospholipids from human heart combining two HPLC methods

The separation of phospholipid classes from human heart was achieved in two steps by high performance liquid chromatography (HPLC) using a silica column with an ultraviolet spectromonitor at 206 nm. A complete partitioning of phosphatidylcholines (PC), phosphatidylethanolamines (PE), phosphatidylinositols (PI), phosphatidylserines (PS), cardiolipins (CL), lysophosphatidylcholines (LPC) and sphingomyelins (Sph) was obtained for further analysis.     

Quantitating heart lipids: Comparison of results obtained using the Iatroscan method with those from phosphorus and gas chromatographic techniques

The precision and accuracy of the Iatroscan method was evaluated by comparing the results obtained with established phosphorus and gas chromatographic techniques. A complete lipid class analysis of rat heart lipids was chosen in order to evaluate the performance of the Iatroscan method for biological samples which contained both neutral lipids and phospholipids. A partial scan and repeat development with chloroform/methanol/water (68.5∶29∶2.5) was introduced to achieve consistently good separations of the phospholipids on the Chromarods in the Iatroscan method. The results showed that the precision of the Iatroscan method for some lipid classes was comparable to that of phosphorus or gas chromatographic techniques, while for other lipid classes it was lower. Compared to the data obtained using the phosphorus method, the Iatroscan data were generally similar, while the gas chromatographic method generally gave lower values. These findings, together with the advantages of time required for analysis, size of sample, and universality of detection, suggest that the Iatroscan is a valuable complementary method for complex lipid analyses.     

An effective separation and purification of rutin and scoparone from Herba Artemisiae Scopariae by solid-phase extraction cartridges packed with an ionic liquid-based silica

Five types of ionic liquid-based silica were synthesized as solid-phase extraction (SPE) sorbents for separation and purification of bioactive compounds, that is, rutin and scoparone extracted from Herba Artemisiae Scopariae. The SilprBImCl material with the highest adsorption capacity was selected as the sorbent for SPE packing. Ethyl acetate and water were found to be suitable washing and eluting solvents, respectively. SilprBImCl was then applied to multi-phase extraction, and its superiority as a sorbent over the commercial cartridge was proven with high rutin and scoparone recoveries of 90.5% and 83.9%, respectively. This highlights the potential of ionic liquid-based silica materials applied to SPE and MPE.     

Preparative separation of phospholipids from soybean by NP-HPLC

Soybean phospholipids-phosphatidylethanolamine (PE), phosphatidylinositol (PI), and phosphatidylcholine (PC)-were separated by normal-phase high performance liquid chromatography. The system was operated in a gradient mode. The experimental variables were gradient time and mobile phase composition. The experimental results showed that PE, PI and PC were resolved by three step-change gradient modes which employed ternary systems of hexane/1-propanol/water (58/40/2 and 56/40/4 by vol.%) and methanol/1-propanol/water (80/18/2, by vol.%) for the gradient times of 10, 30 and 76 min, respectively.     

Erucic acid and phospholipids of newborn rat heart cells in culture

Erucic acid (Δ¹³-docosenoic acid), labeled with¹⁴C in the 1-or 14-position, was incorporated into fetal calf serum and fed to beating, neonatal rat myocardial cells in culture. Uptake of the docosenoic acid during the first 6 hr of incubation was 41 nM/hr/mg protein in 7-day old cells and 29 nM/hr/mg protein in 14-day old cells. Fifty-seven percent of the¹⁴C-activity was taken up from the medium in 24 hr, of which 77% was in the cells and 23% was unaccounted for. Of the¹⁴C-activity taken up, 26% was in extractable lipid, with two-thirds in neutral lipid and one-third in phospholipid. Within the neutral lipid fraction, 88% of the¹⁴C-activity was present in triglycerides; while in phospholipids, 66% of the¹⁴C-activity was in phosphatidylcholine (PC); 14% in phosphatidylethanolamine (PE); 6% in sphingomyelin (SPH) and 1% or less in cardiolipin (DPG). PC had the highest specific activity, followed by SPH and PE. The specific activity of PE was one-half that of SPH when the¹⁴C-erucic acid substrate was labeled at the carboxyl position, but increased to equal that of SPH when the substrate was labeled at the double bond. The fatty acids of PC, PE, and SPH were influenced by erucic acid in the growth medium, but the amounts of each phospholipid were not affected. It is proposed that the altered fatty acid composition associated with incorporation of erucic acid or its metabolites into PC, PE, and SPH may affect integrity and function of heart cell membranes.     

Extraction of phospholipids from plant oils and colorimetric determination of total phosphorus

A rapid extraction procedure to isolate phospholipids from plant oils is described. Glacial acetic acid is used as solvent and the extracted phospholipids are quantified by means of a colorimetric method.     

Response of docosapentaenoic acids of rat heart phospholipids to dietary fat

Groups of male Holtzman strain rats were fed from weanling one of the following diets: 20% hydrogenated soybean fat (20% HF), and 20% HF plus 2%, 3% and 4% corn oil, respectively, for 20 weeks. The animals were killed, and the heart phospholipid fractions isolated by chromatographic procedures. The levels and distribution of the docosapolyenoic acids, especially 22∶5ω3, were compared among the animals fed the corn oil supplemented and nonsupplemented diets. Although dietary linolenate (18∶3ω3) level was very low in the nonsupplemented diet, 22∶5ω3 accounted for 8.4% of the total fatty acids of heart total phospholipids when this diet was fed-half the level of total eicosatetraenoic acids. The amounts of 22∶5ω3 were decreased by corn oil supplementation of the diet and got down to the “normal” range of 2.0–2.5% at corn oil supplementation levels greater than 2%. The docosapolyenoic acids were confined largely to the phosphatidylcholine and phosphatidylethanolamine classes of phospholipids. These findings are discussed from the standpoint of the structural role of the phospholipids in the heart subcellular fractions.     

非硅基介孔材料的合成和研究进展

主要介绍了介孔碳、介孔金属氧化物、介孔金属等几种主要类型的非氧化硅基介孔材料的合成及研究进展,包括合成方法、材料表征以及这些材料的应用等,同时指出这类材料的研究热点和发展趋势.     

In vitro and in vivo effects of exogenous lipids on the enzymatic hydrolysis of rat bile phospholipids

The addition of total phospholipids, phosphatidylcholines, triglycerides, cholesterol or glycerol to incubation media containing rat pancreatic juice and bile labeled with [9,10³H₂] oleic acid (90% of the radioactivity present as phospholipids) had no effect on the hydrolysis of bile endogenous phospholipids. The introduction of 2 or 10 mg of phosphatidylcholines and 0.5 ml of bile (≈ 1.5 mg of phospholipids)into the rat upper duodenum decreased the rate of absorption of rative bile phospholipids. It was not followed by an increase of free fatty acids released from biliary phospholipids in the intestinal lumen. The introduction of bile (0.5 ml) and small amounts of triolein (1.4–3.5 mg) into the duodenum had little effect on the rate of hydrolysis and absorption of native bile phospholipids, but caused a reduced absorption of the free fatty acids released or those coming from initial nonphosphorus biliary lipids. The introduction of bile (0.5 ml) and large amounts of triolein (30 mg) into the duodenum increased the rates of hydrolysis and absorption of endogenous bile phospholipids. These observations suggest that luminal lipid components can modify the organization of luminal micelles and, consequently, the action of the pancreatic phospholipase A₂ and the absorption of bile lipids.     

Enrichment of phospholipids from neutral lipids in peanut oil by high-performance liquid chromatography

Phospholipids from crude peanut oil were enriched on a 2-cm silica column and subsequently separated from neutral lipids within the chromatographic system without prior concentration. Hexane effectively removed the bulk neutral lipids, leaving the adsorbed phospholipids on the silica precolumn. Individual phospholipids were separated from the remaining neutral lipids and from each other by using two mixed solvents and a gradient program. This method separates the phospholipids in approximately 27 min after the desired enrichment level has been reached. The research reported in this paper was a cooperative effort by the Agricultural Research Service of the United States Department of Agriculture and the North Carolina Agricultural Research Service, Raleigh, NC 27695-7625.     

Kinetic study of Fe removal from precipitated silica prepared from yellow phosphorus slag

This purification process may be described by the unreacted shrink core model with solid resultant (inert material) and fixed particle size, which is carried out by the action of nitric acid solution on the precipitated silica obtained from yellow phosphorus slag which was leached with phosphoric acid. The study results indicate that the purification process is a chemical reaction controlling step and its apparent activation energy E_a is 30.354 kJ/mol, with reaction order 0.6746.     

Fatty acid composition of melon seed oil lipids and phospholipids

Fatty acid compositions of crude melon seed oil from two different sources were compared. Melon seeds fromCitrullus vulgaris (syn.C. lanatus) contained phosphatidylcholine (PC), lysophosphatidylcholine (LPC) and phosphatidylserine (PS), whereas melon seeds fromCitrullus colocynthis contained only PC and LPC, but not PS. Analysis of the total lipids revealed that the major fatty acid of the oils was 18:2n-6.Citrullus vulgaris seed oil contained 71.3% andC. colocynthis contained 63.4% of 18:2n-6. The predominant fatty acids in theC. vulgaris PC were 18:2n-6 (32.2%), 18:1n-9 (26.4%) and 16:0 (22.2%), whereas theC. colocynthis PC contained 44.6% of 18:1n-9 as the major fatty acid. The level of monoenes in theC. colocynthis variety (46.2%) was different from theC. vulgaris (27.3%). The major fatty acid in the LPC was 18:1n-9 for both varieties. Notably, theC. colocynthis variety did not contain any PS. The major fatty acids in theC. vulgaris PS were 18:1n-9 (37.9%) and 18:2n-6 (33.7%). Of all the phospholipids, LPC contained the greatest amount of monoenes, 48.6–52.4%.     

Embryonic vs tumor lipids: II. Changes in phospholipids of developing chick brain,heart, and liver

Brain, heart, and liver tissues were excised from embryos and chicks 10, 13, 16, 19, 22, 27, and 53 days after incubation was initiated and the lipids extracted. The quantitative distribution of the phospholipids and the fatty acid composition of the individual phosphatides were determined for each time period. Each tissue exhibited a distinct phospholipid composition that differed from the composition of egg. Elevated concentrations of particular phosphoglycerides that characterize certain mature tissues were observed at the earliest time period. As development progressed, some phospholipid classes in all tissues showed dramatic change, while others remained relatively constant. Brain showed the most stable composition, while the phosphatides of liver were the most dynamic. Each phospholipid class exhibited a characteristic fatty acid profile that was unique for each tissue. All of the phospholipid classes showed a change in fatty acid composition as development progressed, and, in some tissue, the change was dramatic. The fatty acid composition of brain phosphoglycerides showed the least change, while liver showed the greatest fluctuation. Docosahexaenoic acid and, in most cases, arachidonic acid decreased in the phosphoglycerides with increased development. The decrease in docosahexaenoic acid correlated well with the decreasing mitotic indices of heart and liver cells as development progressed. Comparison of observed abnormal lipid patterns between mature and neoplastic tissue with embryonic tissue lipid profiles suggest that some of the observed abnormalities of neoplasms probably are due to changes in lipid metabolism associated with rapidly proliferating cells, whereas other abnormalities appear to be associated with neoplasia.     

Experimental nephrotic syndrome induced in the rat by puromycin aminonucleoside: Hepatic synthesis of neutral lipids and phospholipids from3H-water and3H-palmitate

Experimental nephrotic syndrome (ascites, proteinuria, hypoalbuminemia, and hyperlipidemia) was induced in male Wistar rats by seven daily subcutaneous injections of puromycin aminonucleoside (20 mg/kg). Hepatic lipogenesis from³H-water and³H-palmitate was investigated in nephrotic and pair fed control rats by using liver slices. Total incorporation of³H-water into neutral lipids was higher in nephrotic than in control rats (413±124 vs. 229±46 nmoles/g/hr, p<.01). Among neutral lipids, the major increase was observed for triacylglycerols (106±26 vs. 72±21 nmoles/g/hr, p<.05), cholesteryl esters (3.7±2.1 vs. 1.4±.7 nmoles/g/hr, p<.05) and, above all, for cholesterol (123 ±48 vs. 36±18 nmoles/g/hr, p<.0025). Total incorporation of³H-water into phospholipids as well as incorporation of³H-water into individual phospholipids were not significantly increased. Incorporation of³H-palmitate into neutral lipids was increased (312±84 vs. 221±28 nmoles/g/hr, p<.05). Among neutral lipids, a significant increase was observed for 1,3-diacylglycerols (19±3 vs. 13±3 nmoles/g/hr, p<.025), triacylglycerols (228±50 vs. 163±14 nmoles/g/hr, p<.05) and cholesteryl esters (18±5 vs. 10±1 nmoles/g/hr, p<.01). Incorporation of³H-palmitate into phospholipids was not significantly affected. The difference in hepatic lipogenesis between nephrotic and control rats was even more pronounced if the data were corrected for the total liver weight which was significantly increased in the nephrotic rats (11.3±.3 vs. 8.5±.1 g, p<.001). These findings indicate that the synthesis of neutral lipids from both³H-water and³H-palmitate is elevated in rat with aminonucleoside-induced nephrotic syndrome. The possible role of the increased hepatic lipogenesis in the pathogenesis of the nephortic hyperlipidemia is discussed.     

Rapid synthesis of nanostructured porous silicon carbide from biogenic silica

Nanostructured silicon carbide (SiC) is an exceptional material with numerous applications, for example, in catalysis, biomedicine, high-performance composites, and sensing. In this study, a fast and scalable method of producing nanostructured SiC from plant materials by magnesiothermic reduction via self-propagating high-temperature synthesis (SHS) route was developed. The produced biogenic material possessed a high surface area above 200 m²/g with a SiC crystallite size below 10 nm, which has not been done previously by SHS. This method enables affordable synthesis of the material plant-based precursors in a reaction that only takes a few seconds, thereby paving a way for nanostructured SiC production in high volumes using renewable resources. The material was also functionalized with carboxylic acid and bisphosphonate moieties, and its use as metal adsorbent in applications such as wastewater remediation was demonstrated.     

Studies on the lipids of sheep red blood cells. II. The incorporation of phosphorus into phospholipids of HK and LK cells

The incorporation of inorganic phosphate (as NaH₂PO₄) into the phospholipids of sheep red blood cells was studied in vitro in blood samples from five highpotassium (HK) and five low-potassium (LK) sheep. The erythrocytes from HK sheep incorporated more activity in 4 hr than those from the LK sheep. However no activity was incorporated into the major phospholipids of the cells (phosphatidyl ethanolamine, phosphatidyl serine, and sphingomyelin) of either group. The phosphatidic acid fraction was labeled in both groups and to a significantly greater extent in the HK samples. However the highest activity in the phospholipid of sheep red-cells was located in three unknown compounds not previously detected. Their specific activities were the same in the HK and the LK samples although they were present in slightly larger amounts in the HK samples. In general, incorporation was at a rather low level, and from stoichiometric considerations it was concluded that the metabolism in the red-cell phospholipids could not be directly involved in the active transport of ions across the cell membrane. This work also confirmed a previous report that no quantitative differences exist among the major phospholipid classes in the two types of cells.     

用磷表快速测定磷石膏中水溶磷和非水溶磷

用磷表分析法代替磷钼酸喹啉容量法测定磷石膏中的水溶磷和非水溶磷,分析方法简单,经与传统容量法对照,结果准确、可靠,降低了分析成本,能满足生产控制分析需求。