Objective To investigate the method for the determination of concentration of erucic acid methyl ester in human hepatocellular line BEL-7404.Methods The concentration of erucic acid methyl ester in the blank cell lysate,the medium and the lysate of cells that added erucic acid methyl ester were detected by thin layer chromatography-gas chromatography(TLC-GC),and the relationship between the concentration of erucic acid methyl ester and the peak area.Results A good linear correlation between peak area and concentration were obtained within a concentration range of 50~5 000 μg/ml(r=0.999 95).The amount of erucic acid methyl ester microemulsion got into human hepatocellular line BEL-7404 were 24.9%.Conclusions TLC-GC can determine the concentration of the drug in the cell accurately;this method is simple,rapid and stability.

目的 首次合成未见报道的N,N-二(2-氯乙基)芥酸酰胺,探讨其体内外抗肿瘤活性.方法 ①以芥酸和双(2-氯乙基)胺盐酸盐为原料、吡啶为催化剂合成N,N-二(2-氯乙基)芥酸酰胺,分离纯化后核磁共振谱确证结构;②用N,N-二(2-氯乙基)芥酸酰胺作用于Tca8113,用倒置显微镜镜下观察其对Tca8113细胞的生长状况的影响,PI染色经流式细胞术分析其杀伤效应;③S180小鼠动物肿瘤模型观察N,N-二(2-氯乙基)芥酸酰胺对肿瘤的体内抑制作用.结果 合成出了N,N-二(2-氯乙基)芥酸酰胺并确证了其结构;N,N-二(2-氯乙基)芥酸酰胺于体外0.15～10.0 μg/ml的浓度范围对Tca8113有较强的杀伤效应,杀伤率均超过30%,但与浓度递增无明显的正相关,在100.0 μg/ml以上的浓度反而杀伤效果不佳;体内抑制S180的生长,剂量为5 mg/kg时,抑制作用最好,生命延长率为38.20%,体重平均增长速率为2.45%.结论 采用微型合成法能够快速、方便地合成出N,N-二(2-氯乙基)芥酸酰胺;N,N-二(2-氯乙基)芥酸酰胺在在体内外具有明显的抗肿瘤活性.

Objective To study the synthesis of the N-alcohol erucamide and its activity of anti-tumor in vitro.Methods (1) Through the micro-organic synthesis technology,erucic acid and methanol were coupled with ethanolamine as raw matierals,sodium as catalyst to synthesize the N-alcohol erucamide,isolation and purification achieved by silical gel column chromatography,and Its structure was confirmed by infrared spectra and nuclear magnetic resonance spectrum;(2) BEL-7404 was cultured in media added to the N-ethanol erucamide.The cells's growth was observed in inverted microscope.The effect of inhibiting cancer cells was detected with MTT.Results The N-alcohol erucamide was synthesized successfully;Its strong inhibiting effect was recorded beyond 50.0 μg/ml.The inhibiting rates were all beyond 40%,the inhibitory concentration 50% (IC50) was 101.77588 μg/ml.Conclusion Through the microscale organic synthesis,N-alcohol erucamide can be synthesized quickly and conveniently.N-alcohol erucamide has obvious inhibiting effcet on BEL-7404 cells in vitro.

Cyclohexyl erucate was prepared by the reaction of erucic acid with cyclohexanol in concentrated sulfuric acid in microscale organic laboratory and isolated with column chromatography.Its structure was identified by IR and 1H-NMR.By observing cyclohexyl erucate on tongue Tca8113 the growth of cancer cells,research cyclohexyl erucate on Tca8113 tongue cancer cells in vitro.The results showed that unsaturated fatty acids could be synthesized quickly and conveniently by using microscale organic synthesis.The killing effect of cyclohexyl erucate on tongue cancer cells was significant when more than 500 μg/mL.

Isopropyl erucate was prepared by the reaction of erucic acid with isopropyl alcohol in concentrated sulfuric acid in microscale organic lab and purified with column chromatography. Its structure was characterized by IR and ~1H-NMR.The result showed that isopropyl erucate was synthetized and identified.Unsaturated fatty acids can be synthetized quickly and conveniently with microscale organic synthesis technique.