Quantitation of yeast ceramides using high-performance liquid chromatography–evaporative light-scattering detection

Zhou, Quan; Zhang, Lin; Fu, Xueqi; Chen, Guoqiang

doi:10.1016/s1570-0232(02)00466-x

Cited by 17 publications

(8 citation statements)

References 24 publications

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“…Therefore, separation and quantifi cation of CMH and sphingomyelin from tissues of fish by this HPLC-ELSD method was more accurate and sensitivity than HPLC-UV 17,[20][21][22] . The cyanopropyl cyano column is a special chemically bonded stationary phase column based on silica.…”

Section: Discussionmentioning

confidence: 98%

“…Besides, the evaporative light-scattering detector ELSD for HPLC which is insensitive to the mobile-phase solvents and thus allows direct quantifi cation, has been successfully applied to analyze glycolipids in edible plants 17 , yeast 20 and other foodstuff 21,22 . The aim of this study was to develop an optimal HPLC-ELSD condition for separation, purification and determination of sphingolipids.…”

Section: Introductionmentioning

confidence: 99%

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Rapid Quantitative Analysis of Sphingolipids in Seafood Using HPLC with Evaporative Light-Scattering Detection: Its Application in Tissue Distribution of Sphingolipids in Fish

Duan¹,

Sugawara²,

Hirata³

2010

J. Oleo Sci.

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Sphingolipids are ubiquitous in all eukaryotic organisms and known to be essential constituents of cellular membranes. Recently, various physiological functions of dietary sphingolipids, such as preventing cancer, improving skin barrier and contributing to central nervous system myelination have been demonstrated. To characterize the sphingolipids from fish as food components, tissue distribution of sphingomyelin and glycosylceramide (ceramide monohexoside, CMH) in fi sh were determined in this study. We established a rapid, accurate and effective method for separation, purification and determination of sphingolipids by using high-performance liquid chromatography with evaporative light-scattering detector (ELSD-HPLC). Sphingolipids were extracted and quantified from pacific saury (Cololabis saira). Sphingomyelin in different tissues of Cololabis saira ranged from 2.5 ± 0.2 mg/g to 27.6 ± 2.1 mg/g, the content in brain was the highest, followed by eyes, and CMH contents were less than 23.0 ± 2.4 mg/g in all tissues. These results revealed that fi sh contained CMH and sphingomyelin as same levels as most of the terrestrial organisms and suggested marine organisms could be used as a potential source of precious and useful complex lipids.

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Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Rapid Quantitative Analysis of Sphingolipids in Seafood Using HPLC with Evaporative Light-Scattering Detection: Its Application in Tissue Distribution of Sphingolipids in Fish

Duan¹,

Sugawara²,

Hirata³

2010

J. Oleo Sci.

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“…The pellet containing the membrane fraction was then washed three times with 1 ml of cold PBS, centrifuged, and suspended in 220 l of PBS. Lipid extractions and ceramide determinations from each of above fractions were carried out according to the method of Zhou et al (30). Briefly, a 200-l sample of each fraction was mixed with 4 ml chloroform-methanol (2:1) and extracted for its lipid contents for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Akt/eNOS signaling pathway mediates inhibition of endothelial progenitor cells by palmitate-induced ceramide

Tan

et al. 2015

American Journal of Physiology-Heart and Circulatory Physiology

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Palmitate (PA) impairs endothelial progenitor cells (EPCs). However, the molecular mechanism underlying the suppressive function of PA remains largely unknown. Ceramide, a free fatty acid metabolite, mediates multiple cellular signals. We hypothesized that ceramide acts as an intermediate molecule to mediate inhibition of EPCs by PA. We first demonstrated that PA could inhibit the attachment, migration, and tube formation of EPCs through suppression of the Akt/endothelial nitric oxide (NO) synthase (eNOS) signaling pathway. In addition, we observed that PA could induce ceramide accumulation in EPCs. To test whether the accumulation of ceramide causes EPC dysfunction, the ceramide synthesis inhibitors myriocin and fumonisin B1 were used. We that found both inhibitors could effectively abolish PA-mediated EPC inhibition. Furthermore, the ceramide deacylation inhibitor N-oleoylethanolamine could augment the inhibitory effect of PA on EPCs, indicating that it is ceramide, not its metabolites, that mediates the suppression of EPCs by PA. We have previously shown that Akt/ eNOS phosphorylation was reduced after PA treatment, which, in turn, hampered the normal bioavailability of NO, leading to impaired functions of EPCs. To test the role for ceramide in this process, a clinically used NO donor, sodium nitroprusside, was used. We found that sodium nitroprusside could rescue the suppressive effects of ceramide on EPCs, suggesting that ceramide-mediated EPC inhibition might be through reduction of NO production. Taken together, our findings indicated that ceramide-induced reduction of NO might be the molecular mechanism for PA-mediated EPC inhibition; thus, targeting either ceramide or NO production might be an effective means for improvement of EPC functions in diseases.

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“…This detection method has been applied effectively for at least the last two decades to quantify a wide spectrum of natural and synthetic compounds including pharmaceuticals (i.e., drug molecules and excipients including polymers, sugars, lipids, and surfactants) (4,24,(26)(27)(28)(29), natural products and biologics (30,31). foods, and beverages (32,33). Additionally, size exclusion chromatography (SEC) has been used to resolve polymers based on differences in molecular size where separation occurs as a result of pore size of the packing material (11).…”

Section: Introductionmentioning

confidence: 99%

Development of a Robust Method for Simultaneous Quantification of Polymer (HPMC) and Surfactant (Dodecyl β-D-Maltoside) in Nanosuspensions

Patel

Bummer

2015

AAPS PharmSciTech

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Abstract. This report describes the development of a chromatographic method for the simultaneous quantification of a polymer, hydroxypropyl methylcellulose (HPMC), and a surfactant, dodecyl β-Dmaltoside (DM), that are commonly used in the physical stabilization of pharmaceutical formulations such as nanosuspensions and solid dispersions. These excipients are often challenging to quantify due to the lack of chromophores. A reverse phase size exclusion chromatography (SEC) with evaporative light scattering detector (ELSD) technique was utilized to develop an accurate and robust assay for the simultaneous quantification of HPMC and DM in a nanosuspension formulation. The statistical design of experiments was used to determine the influence of critical ELSD variables including temperature, pressure, and gain on accuracy, precision, and sensitivity of the assay. A robust design space was identified where it was determined that an increase in the temperature of the drift tube and gain of the instrument increased the accuracy and precision of the assay and a decrease in the nebulizer pressure value increased the sensitivity of the assay. In the optimized design space, response data showed that the assay could quantify HPMC and DM simultaneously with good accuracy, precision, and reproducibility. Overall, SEC-ELSD proved to be a powerful technique for the simultaneous quantification of HPMC and DM. This technique can be used to quantify the amount of HPMC and DM in nanosuspensions, which is critical to understanding their effects on the physical stability of nanosuspensions.KEY WORDS: design of experiments; dodecyl β-D-maltoside; evaporative light scattering detector; HPMC; nanosuspensions.

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Quantitation of yeast ceramides using high-performance liquid chromatography–evaporative light-scattering detection

Cited by 17 publications

References 24 publications

Rapid Quantitative Analysis of Sphingolipids in Seafood Using HPLC with Evaporative Light-Scattering Detection: Its Application in Tissue Distribution of Sphingolipids in Fish

Rapid Quantitative Analysis of Sphingolipids in Seafood Using HPLC with Evaporative Light-Scattering Detection: Its Application in Tissue Distribution of Sphingolipids in Fish

Akt/eNOS signaling pathway mediates inhibition of endothelial progenitor cells by palmitate-induced ceramide

Development of a Robust Method for Simultaneous Quantification of Polymer (HPMC) and Surfactant (Dodecyl β-D-Maltoside) in Nanosuspensions

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