目的：建立高效液相色谱-质谱联用测定人血浆中二甲氨基乙醇（DMAE）浓度的方法。方法：采用 API 3000 型 LC-MS/MS 液质联用仪,Phenomenex Ultimate C18 色谱柱（50 mm×3.0 mm,5 µm）,流动相为甲醇-水（80∶20）（含 0.1% 甲酸）,流速 0.3 mL·min-1。样品在碱性条件下用乙酸乙酯漩涡萃取,定量转移出乙酸乙酯层后再用 0.1% 的甲酸水溶液反萃,最后取酸性水溶液进样。以 MRM 模式检测,外标法 定量。结果：DMAE 的保留时间为 2.79 min,在 3.125~500 ng·mL-1浓度范围内线性关系良好,定量下限为 3.125 ng·mL-1,日内、日间 RSD 分别小于 4.8% 和 6.0%,方法回收率为 98.7%~101.9%,萃取回收率为 32.5%~34.5%。21 位健康受试者交叉服用 2 种胶囊制剂,在单次口服 300 mg 盐酸甲氯酚酯后,采用本方 法测定血浆中 DMAE 经时浓度,比较 2 种制剂的药代动力学参数并评价其相对生物利用度。试验制剂和 参比制剂的 Cmax 分别为（277.66±107.25）、（266.18±118.37）µg·L-1,AUC0-t 分别为（721.05±172.72）、（736.03±216.90）h·µg·L-1,AUC0-∞分别为（776.17±203.89）、（835.52±269.99）h·µg·L-1,2 种制剂生物等效。结论：本法具有快速、简便、灵敏、准确等特点,适用于血浆中 DMAE 的浓度测定及药代动力学、生物利用度研究。
Objective：To establish an LC-MS/MS method for determination of 2-dimethylaminoethanol（DMAE） in human plasma. Methods：The assay was conducted with an API 3000 LC-MS/MS system consisting of a Phenomenex Ultimate C18 column（50 mm×3.0 mm,5 μm）. The mobile phase of methanol-water（80∶20,with 0.1% formic acid）was pumped at 0.3 mL·min-1 through the column. The alkalinized sample was extracted with ethyl acetate vortex,quantitatively transferred out of the organic layer to another tube and was re-extracted by 0.2 mL water（with 0.1% formic acid）. The acidic water was finally injected onto the column. Use MRM mode to detect. Results：The retention time of DMAE was 2.79 min. A linearity of DMAE curve was within 3.125-500 ng·mL-1. The lowest concentration of detection in plasma was 3.125 ng·mL-1,the intra-day RSD and the inter-day RSD was less than 4.8% and 6.0% respectively;the method recovery was 98.7%-101.9%;the extract efficiency was 32.5%-34.5%. 21 healthy subjects received single dose of two capsule preparations under a cross-over design. After orally taking 300 mg meclofenoxate hydrochloride,the plasma concentration of DMAE was measured by this method. The main pharmacokinetic parameters were as follows：the Cmax were（277.66±107.25）and（266.18±118.37）µg·L-1,AUC0-t were（721.05±172.72）and（736.03±216.90）h·µg·L-1,AUC0-∞ were（776.17±203.89）and（835.52±269.99）h·µg·L-1. The two capsule preparations were bioequivalence. Conclusion：The method is found to be simple,rapid,sensitive. It is qualified for determination of DMAE in human plasma.
[1] 马春红. 氯酯醒胶囊治疗血管性痴呆的有效性及药理分析[J]. 中国医药指南,2014,12(12):82
[2] 朱玲. 氯酯醒抗衰老作用的药理解析[J].中国现代医药杂志,2007,9(6):127
ZHU L.Pharmacological analysis of the anti-aging effect of meclofenoxate[J].Mod Med J China,2007,9(6):127
[3] 毛蕾,兰志勋,马德祥.盐酸甲氯芬酯对老年女性患者全麻后复苏以及早期认知功能的影响[J].实用医院临床杂志,2019,16
(2):57
4 MAO L,L AN ZX ,M A DX .The effect of meclofenoxate hydrochloride on recovery and early postoperative cognitive dysfunction of elder female patients under general anesthesia[J]. Pract J Clin Med,2019,16(2):57
[4] 马建. 盐酸甲氯芬酯治疗重型颅脑损伤的效果观察及对 GCS 评分的影响[J].中国合理用药探索,2018,15(12):143
MA J.Observation on therapeutic effect of meclofenoxate hydrochloride on severe traumatic brain injury and its effect on GCS scores[J].Chin J Ration Drug Use,2018,15(12):143
[5] 蒲友华,刘定远.注射用盐酸甲氯芬酯治疗新生儿缺氧缺血性脑病临床研究[J].儿科药学杂志,2008,14(4):47
PU YH,LIU DY.The clinical research of meclofenoxate in the treatment of neonatal hypoxic-ischemic encephalopathy[J].J Pediatr Pharm,2008,14(4):47
[6] 陆磊,孙晓江,张进,等.盐酸甲氯芬酯治疗急性脑出血的疗效评价[J].药学服务与研究,2007,7(4):271
LU L,S UN XJ ,Z HANG J ,e t al .T he clinical efficacy of meclofenoxate hydrochloride in treatment of patients with acute intracerebral hemorrhage[J].Pharm Care Res,2007,7(4): 271
[7] 付剑亮,孙晓江,赵玉武,等.氯酯醒治疗血管性痴呆的有效性和安全性研究[J].中国现代医学杂志,2007,17(13):1615
FU JL,SU N XJ,Z HAO YW ,e t al .Sa fety and efficacy of meclofenoxate in treatment of patients with vascular dementia[J]. Chin J Mod Med,2007,17(13):1615
[8] 牛超,惠延年,王雨生,等.盐酸甲氯芬酯对人视网膜色素上皮细胞增生的影响[J].眼科新进展,2006,26(4):245
NIU C,HUI YN,WANG YS,et al.Influence of meclofenoxate hydrochloride on proliferation of human retinal pigment epithelial cells[J].Recent Adv Ophthalmol,2006,26(4):245
[9] ZOU J J ,JIH J ,WU DW ,et al ．Bioequivalence and pharmacokinetic comparison of a single 200-mg dose of meclofenoxate hydrochloride capsule and tablet formulations in healthy chinese adult male volunteers：a randomized sequence, open-label,two-period crossover study[J]．EM Inc USA,2008, 30(9)：165
[10] 华卡,李健和,朱运贵,等.盐酸甲氯芬酯胶囊在健康人体内的生物等效性研究[J].中国药房,2011,22(18):1669
HUA K,LI JH,ZHY YG,et al .Bioequivalence study of meclofenoxate hydrochloride capsule in healthy volunteer[J].J China Pharm,2011,22(18):1669
[11] 荆莉,于翠霞,张倩,等.盐酸甲氯芬酯胶囊在健康受试者体内血药浓度测定及生物等效性研究[J].中国药师,2012,15(6): 800
JIN L,YU CX,ZHANG Q,et al.Determination of meclofenoxate hydrochloride capsules in human plasma by HPLC and bioequivalence study[J].China Pharm,2012,15(6):800
[12] 石玲子,南峰,郭之晨,等.反相高效液相色谱法测定人血浆中对氯苯氧乙酸的含量[J].现代预防医学,2009,36(4):722
SHI LZ,NAN F,GUO ZC,et al .Determination of 4-chlorophenoxyacetic acid in human serum by reversed phase high performance liquid chromatography[J].Mod Prev Med,2009,36(4):722
[13] MONEEB M,ELGAMMAL F,SABRY S.Development and validation of a stability indicating HPLC-diode array-fluorescence method for the determination of meclofenoxate hydrochloride and p-chlorophenoxyacetic acid[J].J Appl Pharm,2016,6(7):1
[14] 周卫民,李健和,黎银波,等.盐酸甲氯芬酯分散片含量测定的方法比较[J].中国医药导报,2009,6(11):49
ZHOU WM,LI JH,LI YB,et al.Methods comparison of the determination of meclofenoxate hydrochloride dispersible tablet[J]. Chin Med Herald,2009,6(11):49
[15] 贺凌云,夏宏鹏,吴建平,等.高效液相色谱法测定盐酸甲氯芬酯胶囊的含量[J].分析试验室,2007,26(9):74
HE LY,XIA HP,WU JP,et al.Determination of meclofenoxate hydrochloride capsules by HPLC[J].Anal Lab,2007,26(9):74
[16] 中华人民共和国药典 2015 年版.四部:9012 生物样品定量分析方法验证指导原则华人民共和国药典 2015 年版.四部:9012 生物样品定量分析方法验证指导原则[S].2015:363
ChP 2015.Vol IV:9012:Verification Guidelines for Quantitative Analysis Methods of Biological SampleshP 2015.Vol IV:9012:Verification Guidelines for Quantitative Analysis Methods of Biological Samples[S].2015:363
