Ledger Robin
School of Pharmacy, University of Otago, P.O. Box 913, Dunedin, New Zealand.
J Biochem Biophys Methods. 2003 Aug 29;57(2):105-14. doi: 10.1016/s0165-022x(03)00099-x.
Less than 11% of the dose of bupivacaine could be accounted for in urine from 10 patients receiving continuous epidural infusions. HPLC analysis of metabolites confirmed (S)-bupivacaine was more extensively metabolised than (R)-bupivacaine, and dealkylation was the predominant metabolic pathway although co-elution of metabolites made quantitation difficult. The percentage of (S)-2',6'-pipecoloxylidide and co-eluting metabolites excreted relative to (R)-2',6'-pipecoloxylidide from three patients was 0.32+/-0.05, while for seven patients it was 1.28+/-0.09. Conversely, the percentage of (S)-3'-hydroxy bupivacaine and co-eluants excreted relative to (R)-2',6'-pipecoloxylidide from the three patients (1.76+/-0.48) was greater than the seven patients (0.19+/-0.09). Urinary metabolites were analysed for evidence of aliphatic hydroxylation of bupivacaine. Chiral liquid chromatography-mass spectrometry (LC-MS) on an alpha(1)-glycoprotein column at pH 7 used hydroxylamine acetate as the volatile mobile phase. Compounds tentatively identified as hydroxybupivacaines by MRM were verified by their product ion spectra in a subsequent MS-MS run. Eighteen oxygenated metabolites of bupivacaine were detected, half of which were hydroxylated on nonaromatic groups. Equal numbers of mono- and dihydroxybupivacaines were excreted. There was no evidence to suggest the presence of (S)-4'-hydroxybupivacaine, 2'-hydroxymethylbupivacaine, 3'-hydroxy-2',6'-pipecoloxylidide or a piperidone. The metabolite previously identified as (S)-4'-hydroxybupivacaine was not hydroxylated on the xylyl group.
在接受连续硬膜外输注的10名患者的尿液中,布比卡因的剂量占比不到11%。代谢物的高效液相色谱分析证实,(S)-布比卡因比(R)-布比卡因代谢更广泛,脱烷基化是主要代谢途径,尽管代谢物的共洗脱使得定量分析困难。三名患者中,相对于(R)-2',6'-哌啶甲酰苯胺,(S)-2',6'-哌啶甲酰苯胺及共洗脱代谢物的排泄百分比为0.32±0.05,而七名患者的该值为1.28±0.09。相反,三名患者中,相对于(R)-2',6'-哌啶甲酰苯胺,(S)-3'-羟基布比卡因及共洗脱物的排泄百分比(1.76±0.48)高于七名患者(0.19±0.09)。分析尿液代谢物以寻找布比卡因脂肪族羟基化的证据。在pH 7的α(1)-糖蛋白柱上进行手性液相色谱-质谱联用(LC-MS)分析,使用乙酸羟胺作为挥发性流动相。通过多反应监测(MRM)初步鉴定为羟基布比卡因的化合物,在随后的串联质谱(MS-MS)运行中通过其产物离子谱进行了验证。检测到18种布比卡因的氧化代谢物,其中一半在非芳香族基团上发生了羟基化。单羟基布比卡因和二羟基布比卡因的排泄量相等。没有证据表明存在(S)-4'-羟基布比卡因、2'-羟甲基布比卡因、3'-羟基-2',6'-哌啶甲酰苯胺或哌啶酮。先前鉴定为(S)-4'-羟基布比卡因的代谢物在二甲苯基上未发生羟基化。