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首页> 外文期刊>Biologicals: Journal of the International Association of Biological Standardization >Quantification of lipopolysaccharides in outer membrane vesicle vaccines against meningococcal disease. High-performance liquid chromatographic determination of the constituent 3-hydroxy-lauric acid.
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Quantification of lipopolysaccharides in outer membrane vesicle vaccines against meningococcal disease. High-performance liquid chromatographic determination of the constituent 3-hydroxy-lauric acid.

机译：定量针对脑膜炎球菌疾病的外膜囊泡疫苗中的脂多糖。高效液相色谱法测定组成的3-羟基月桂酸。

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摘要

A high-performance liquid chromatographic (HPLC) assay for quantification of lipopolysaccharides (LPSs, endotoxins) in outer membrane vesicle vaccines against meningococcal disease has been developed. The LPS constituent, 3-hydroxy-lauric acid, served as marker substance for the quantification. LPS from the vaccine was precipitated by ethanol and the fatty acid constituents, including 3-hydroxy-lauric acid, were released by acidic hydrolysis, collected and purified by solid phase extraction on C18 disc-cartridges and converted into phenacyl esters for UV detection at 240 nm. Quantification of the derivatized 3-hydroxy-lauric acid was achieved by HPLC using a Brownlee RP-18 reversed phase column with acetonitrile/water (68:32, v/v) as mobile phase. The method was found to be linear over the range 3-49 microg LPS/ml with a sensitivity of 1.6 (microg/ml)(-1). The repeatability (within-day precision) of the method at three levels (3-49 microg LPS/ml) was 6-14% relative standard deviation and the intermediate (between-day) precision was 7% relative standard deviation (at level 15 microg LPS/ml). The method has been successfully used in the quality control of a meningococcal B outer membrane vesicle vaccine, containing 4-8% LPS relative to protein (w/w), in our laboratory for three years. Copyright 2002 The International Association for Biologicals.
机译：已开发出一种高效液相色谱（HPLC）测定方法，用于定量抗脑膜炎球菌病的外膜囊泡疫苗中的脂多糖（LPS，内毒素）。 LPS成分3-羟基月桂酸用作定量的标记物。疫苗中的LPS在乙醇中沉淀，脂肪酸成分（包括3-羟基月桂酸）通过酸性水解释放出来，收集并通过固相萃取在C18圆盘盒上纯化，并转化为苯甲酸酯，以便在240进行紫外线检测纳米使用Bownlee RP-18反相色谱柱，以乙腈/水（68:32，v / v）作为流动相，通过HPLC对衍生化的3-羟基月桂酸进行定量。发现该方法在3-49微克LPS / ml范围内是线性的，灵敏度为1.6（微克/ ml）（-1）。在三个级别（3-49微克LPS / ml）下，该方法的重复性（日精度）为相对标准偏差的6-14％，中间（日间）精度为相对标准偏差的7％（15级）微克LPS /毫升）。在我们的实验室中，该方法已成功用于质量控制为相对于蛋白质（w / w）为4-8％LPS的脑膜炎球菌B外膜囊泡疫苗的质量控制，为期三年。版权所有2002国际生物制品协会。

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