• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.1
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• pp.47-51
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• 2004

Ceramide III was prepared by the cultivation of Saccharomyces cerevisiae. Ceramide III was partitioned from the cell extracts by solvent extraction and analyzed by Normal Phase High Performance Liquid Chromatography (NP-HPLC) using Evaporative Light Scattering Detector (ELSD). We experimentally determined the mobile phase composition to separate ceramide III with NP-HPLC. Three binary mobile phases of n-hexane/ethanol, n-hexane/lsoprophyl Alcohol(IPA) and n-hexane/n-butanol and one ternary mobile phase of n-hexane/IPA/methanol were demonstrated. For the binary mobile phase of n-hexane/ethanol, the first mobile phase composition, 95/5(v/v), was step-increased to 72/23(v/v) at 3 min. In the binary mobile phase, the retention time of ceramide III was 7.87min, while it was 4.11 min respectively in the ternary system, where the mobile phase composition of n-hexane/IPA/methanol, 85/7/8(v/v/v), was step-increased to 75/10/15(v/v/v) at 3 min. However, in the ternary mobile phase, the more peak area of ceramide III was observed.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.3 s.47
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• pp.419-426
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• 2004

The nano-ceramide capsulation is a technique that capsulates ceramide III and tocopheryl linoleate at the mono-vesicle to act on the horny layer in skin. In this technique, $0.5{\~}5.0\;wt\%$ of hydrogenated lecithin and $0.01{\~}2.00\;wt\%$ of lysolecithin are used as the membrane-strengthen agents of the mono-vesicle and $5.0{\~}10.0\;wt\%$ of propylene glycol and $5.0{\~}10.0\;wt\%$ of ethyl alcohol are used as solvents. Active ingredients such ceramide III and tocopheryl linoleate are utilized to enhance the moisturizing efficacy and treat atopy skin. These materials do not contain synthetic emulsifiers. The optimal conditions or nano-ceramide capsulation are such that particles pass Microfludizdizer 3 times at 1,000 bar and $60{\~}70^{\circ}C$ and pH of nano capsules is $5.8{\pm}0.5.$ The average size of particles is $63.1{\pm}7.34\;nm$ showing lucid state like water by the laser light scattering. A zeta potential value is $-55.1\pm0.84\;mV.$ Through clinical tests, the moisturizing effect (in-vivo, n=8, p-value<0.05) showed $21.15\%$ of improvement comparison to comparison-samples and $36.31\%$ of improvement compared to the state before treatment. Moreover, the effectiveness of atopy skin showed positive reaction from 10 volunteers.

• Proceedings of the SCSK Conference
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• 2003.09b
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• pp.268-279
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• 2003

The nano capsulation of the ceramide was a technique that capsulated ceramide III and tocopheryl linoleate at the mono-vesicle, so as to act the horny layer in skin. It was used 0.5-5.0 wt% of hydrogenated lecithin and 0.01~2.00 wt% of lysolecithin as the membrane-strengthen agents of the mono-vesicle, 5.0~10 wt% of propylene glycol and 5.0~10.0 wt% of ethyl alcohol made by high-pressure Microfluidizer. To enhance the moisturizing efficacy and treat an atopy skin, used ceramide III and tocopheryl linoleate as the active ingredients, and it was made the nano-capsule that synthetic emulsifiers were free. The optimal condition of capsulation of nano ceramide was as follows. The conditions were 3 times at 1,000bar and 60-7$0^{\circ}C$. The particle size showed 63.1$\pm$7.34 nm such as the transparence water as the results for measuring by the laser light scattering. A zeta potential value was -55.1$\pm$0.84 ㎷. The result of the clinical test, the moisturizing effect (in-vivo, n=8, p-value<0.05) was improved 21.15% compared to control, as well as it was improved 36.31 % before the treatment. Moreover, the effectiveness of atopy skin indicated positive reaction that patients were 10 volunteers.

• Applied Chemistry for Engineering
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• v.21 no.6
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• pp.603-609
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• 2010

This paper aims to investigate the lyotropic behaviors of DSPC and CER3 when they are swollen by GLY as a solvent. The analyses were carried out on DSC, XRDs, PM, and Cryo-SEM. CER3 which has its high crystallinity and structural similarity with DSPC was well arranged up to 7.0 wt% in comparison to 20 wt% DSPC without any separation, but it was separated from the liquid crystalline (LC) phase to form another crystalline phase with the expression of its characteristic peak in XRDs and eutectic thermal behavior in DSC. Introducing CER3, two types of patterns were shown in XRD spectra; one is SPP expressed in a normal LC and another is LPP expressed in human skin SC. Therefore, it was confirmed that the incorporation of CER3 makes LC structure more similar to human skin. In Cryo-SEM study, it was shown that CER3 makes LC structure thicker and denser.

• Bulletin of the Korean Chemical Society
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• v.28 no.6
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• pp.1021-1030
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• 2007

The effects of the creamide III (CER3) and cholesterol (CHOL) on the structure of a non-hydrous distearoyl phosphatidylcholine (DSPC)-based lamellar liquid crystal (LC) hydrated by only propylene glycol (PG) without water were investigated by differential scanning calorimetry (DSC), X-ray diffractions (XRDs), and polarized microscope (PM). As soon as CER3 was incorporated into the lamellar phase, the characteristic LPP was appeared as well as the characteristic SPP, and the formation of separated CER3 crystalline phase was observed depending upon the increase of CER3 content by XRDs. Also, by DSC, it was shown that the increase of CER3 made the monotectic thermal transition be changed to the eutectic thermal transition which indicates the formation of separated CER3 crystalline phases and the main transition temperatures (Tc1) to be gradually decreased and the enthalpy change (ΔH) to be linearly increased. Incorporating CHOL, the formation of LPP and SPP showed almost similar behaviors to CER3, but incorporating small amounts of CHOL showed the characteristic peaks of CHOL which meant the existence of crystalline CHOL phase due to the immiscibility of CHOL with DSPC swollen by PG differently from CER3, and increasing CHOL made the intensity of the 1st order diffraction for LPP weakened as well as the intensities of the characteristic diffractions for DSPC. Also, in the results of DSC, it showed more complex thermal behaviors having several Tc than CER3 due to its bulky chemical structure. In the present study, the inducement of CER3 and CHOL as other lipids present in human stratum corneum (SC) into a non-hydrous lamellar phase is discussed in terms of the influence on their structural and thermal transition.

• Archives of Pharmacal Research
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• v.24 no.2
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• pp.136-143
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• 2001

Fumonisins are specific inhibitors of ceramide synthase in sphingolipid metabolism. An alteration in sphingolipid metabolism as a result of fumonisin exposure is related to cell death (Yoo et al., 1992). The objective of this study was to investigate whether elevated free sphinganine levels are related to the sensitivity of cultured cells to fumonisin exposure. Fumonisin $B_1$ elevated the intracellular free sphinganine concentraions in both LLC-$PK_1$ and Chinese hamster ovary (CHO) cells. However, CHO cells are resistant to fumonisin cytotoxicity at 50${u}m$, while LLC-$PK_1$ cells are sensitive at concentrations greater than 357M. The intracellular concentration of free sphinganine in LLC-$PK_1$ cells treated at 50${u}m$ fumonisin $B_1$ for 72 h was approximately 1450 pmol/mg protein relative to the 37 pmol observed in the control culture. Under the same conditions, the population of apoptotic cells in the 50${u}m$ fumonisin $B_1$-treated culture was approximately 37% of the total compared to 12% in the control. The caspase III-like activity after 72 h in the 50${\mu}$M fumonisin $B_1$-exposed culture Increased to approximately 50 $pmol/mg$ protein/hr compared to 6 $pmol/mg$ protein/hr in the control. L-cycloserine, a serine palmitoyltransferase inhibitory reduced the fumonisin $B_1$-stimulated caspase III-like activity down to the control level. Under the same culture conditions, the intracellular concentration of free sphinganine after-cycloserine plus fumonisin $B_1$ treatment was 140 pmol/mg protein compared to 1450 $pmol/mg$ protein in fumonisin $B_1$ alone. The intracellular concentration of free sphinganine in CHO cells treated with 50${u}m$ fumonisin $B_1$ for 72 h was al)proximately 460 pmol/mg protein, indicating that the mass amount of elevated free sphinganine in the CHO cells was about 32% of that in LLC-$PK_1$ cells. Adding exogenous sphinganine to the CHO cells along with 50${u}m$ fumonisin $B_1$ treatment for 72 h caused both necrosis and apoptosis. In conclusion, the elevated endogenous sphinganine acts as a contributing factor to the fumonisin-induced cell death.