Das A K, Hajra A K
Biochim Biophys Acta. 1984 Nov 14;796(2):178-89. doi: 10.1016/0005-2760(84)90346-1.
Chemical and enzymatic methods have been developed to measure small quantities (10(-8) - 10(-10) mol) of acyldihydroxyacetone phosphate in animal tissues. Lipids extracted from tissue samples with acidic CHCl3/methanol were subjected to solvent partitioning at two different pH values for partial purification of this keto-lipid from other lipids. This lipid was then estimated radiometrically either by chemical reduction with NaB3H4 or by enzymatic reduction with [4B-3H]NADPH using a partially purified acyldihydroxyacetone-phosphate reductase (EC 1.1.1.101). Thin-layer chromatography revealed the presence of a number of 3H-labeled lipids in the NaB3H4-reduced product and further purification of the product was necessary to estimate the amount of acyl[2-3H]glycerol 3-phosphate formed. The enzymatic reduction was very specific for acyl/alkyldihydroxyacetone phosphate. The amounts (nmol/g) of these keto-lipids estimated in different tissues by the enzymatic method were 10.06 +/- 0.64 (guinea pig liver), 4.3 +/- 0.15 (rat liver), 2.1 (rat testis), 1.5 (rad kidney) and 1.2 (rat brain). Monoacylglycerol 3-phosphate, i.e., lysophosphatidic acid, which was co-purified with acyldihydroxyacetone phosphate, was found to be present in relatively larger amounts in tissues. The amounts (nmol/g) of this lipid, estimated by enzymatically measuring the amounts of sn-glycerol 3-phosphate released after alkaline methanolysis of the partially purified lipid extracts, were 143 (guinea pig liver), 58 (rat liver), 53 (rat kidney) and 92 (rat brain). Stearic acid (18:0) was found to be the major (65%) fatty acid present in the lysophosphatidate purified from guinea pig liver.
已经开发出化学和酶法来测量动物组织中少量（10⁻⁸ - 10⁻¹⁰摩尔）的酰基二羟基丙酮磷酸。用酸性氯仿/甲醇从组织样品中提取的脂质在两个不同的pH值下进行溶剂分配，以便从其他脂质中部分纯化这种酮脂质。然后，通过用NaB₃H₄进行化学还原或使用部分纯化的酰基二羟基丙酮磷酸还原酶（EC 1.1.1.101）用[4B-³H]NADPH进行酶还原，以放射性测量法估算这种脂质。薄层色谱显示在NaB₃H₄还原产物中存在多种³H标记的脂质，并且需要对产物进行进一步纯化以估算形成的酰基[2-³H]甘油3-磷酸的量。酶还原对酰基/烷基二羟基丙酮磷酸具有高度特异性。通过酶法在不同组织中估算的这些酮脂质的量（nmol/g）分别为10.06±0.64（豚鼠肝脏）、4.3±0.15（大鼠肝脏）、2.1（大鼠睾丸）、1.5（大鼠肾脏）和1.2（大鼠大脑）。发现与酰基二羟基丙酮磷酸共纯化的单酰甘油3-磷酸，即溶血磷脂酸，在组织中的含量相对较高。通过酶法测量部分纯化的脂质提取物碱性甲醇解后释放的sn-甘油3-磷酸的量来估算这种脂质的量（nmol/g），分别为豚鼠肝脏143、大鼠肝脏58、大鼠肾脏53和大鼠大脑92。发现硬脂酸（18:0）是从豚鼠肝脏纯化的溶血磷脂酸中存在的主要（65%）脂肪酸。
