Qureshi N, Takayama K, Ribi E
J Biol Chem. 1982 Oct 10;257(19):11808-15.
Endotoxin extracted from the heptose-less mutant of Salmonella typhimurium was hydrolyzed in 0.1 N HCl in methanol/water (1:1, v/v) at 100 degrees C to yield lipid A, which was then fractionated on a Sephadex LH-20 column to yield a major monophosphoryl lipid A fraction. The monophosphoryl lipid A was further fractionated by preparative thin layer chromatography. This process yielded three major bands (TLC-1, -3, and -5) and two minor bands (TLC-7 and -9). The purity of these fractions was established by ion exchange and reverse phase high performance liquid chromatography. The thin layer fractions were analyzed by fast atom bombardment mass spectrometry. TLC-1 and -3 gave molecular ions (M-H)- at m/e 1730 and 1716, respectively. Both of these fractions contained beta-hydroxymyristic, lauric, and 3-myristoxymyristic acids in O-acyl linkages. The molecular formula and Mr of TLC-1 are C95H179O22N2P and 1731.16; those of TLC-3 are C94H177O22N2P and 1717.15. TLC-1 was a methyl homolog of TLC-3. The major component of TLC-5 (C80H151O22N2P and Mr = 1506.99) gave a molecular ion at m/e 1506 and contained two beta-hydroxymyristic acids and a lauric acid in the O-acyl linkages. The major component of TLC-7 (C66H125O19N2P and Mr = 1280.83) and the single component of TLC-9 gave molecular ions at m/e 1280 and 1098, respectively. TLC-7 contained lauric and beta-hydroxymyristic acids in the O-acyl linkages. TLC-9 (C54H103O18N2P and Mr = 1098.69) contained a single O-acylated beta-hydroxymyristate group. TLC-1 and -3 were nontoxic in the chick embryo lethality test and regressed established tumors in the syngeneic guinea pigs.
从鼠伤寒沙门氏菌的庚糖缺陷型突变体中提取的内毒素，在100℃下于甲醇/水（1:1，v/v）的0.1N盐酸中水解，得到脂质A，然后在葡聚糖LH - 20柱上进行分离，得到主要的单磷酸化脂质A组分。单磷酸化脂质A通过制备型薄层色谱进一步分离。该过程产生了三个主要条带（TLC - 1、- 3和- 5）和两个次要条带（TLC - 7和- 9）。这些组分的纯度通过离子交换和反相高效液相色谱确定。薄层组分通过快原子轰击质谱分析。TLC - 1和- 3分别在m/e 1730和1716处给出分子离子（M - H）-。这两个组分在O - 酰基连接中都含有β - 羟基肉豆蔻酸、月桂酸和3 - 肉豆蔻氧基肉豆蔻酸。TLC - 1的分子式和Mr分别为C95H179O22N2P和1731.16；TLC - 3的分子式和Mr分别为C94H177O22N2P和1717.15。TLC - 1是TLC - 3的甲基同系物。TLC - 5的主要成分（C80H151O22N2P，Mr = 1506.99）在m/e 1506处给出分子离子，并且在O - 酰基连接中含有两个β - 羟基肉豆蔻酸和一个月桂酸。TLC - 7的主要成分（C66H125O19N2P，Mr = 1280.83）和TLC - 9的单一成分分别在m/e 1280和1098处给出分子离子。TLC - 7在O - 酰基连接中含有月桂酸和β - 羟基肉豆蔻酸。TLC - 9（C54H103O18N2P，Mr = 1098.69）含有一个单一的O - 酰化β - 羟基肉豆蔻酸基团。TLC - 1和- 3在鸡胚致死试验中无毒，并且能使同基因豚鼠体内已形成的肿瘤消退。
