| Objectives:1. To construct an ultra performance liquid chromatography （UPLC） method to detect the serum concentration of levetiracetam （LEV）.2. To construct an UPLC method to detect the serum concentration of active metabolite of oxcarbazepine 10,11-dihydro-10-hydroxyl carbamazepine（MHD）.3. To observe and analyze the correlation of plasma LEV concentration with clinical efficacy in patients with epilepsy.4. To observe and analyze the correlation of plasma MHD concentration with clinical efficacy in patients with epilepsy.Methods:1. Serum LEV concentration was detected using the constructed UPLC-UV method.Aminophylline was served as the internal standard, and plasma sample was injected for analysis after protein precipitation with perchloric acid. Detection conditions were shown as follows: chromatographic column: 50 mmx2.1 mm ACQUITY UPLCBEH C18 1.7 μm chromatographic column （Waters, USA）; ACQUITY UPLC column on-line filtration system （Waters, USA）; the detection wave length was 210nm; flow velocity was 0.15 mL·min-1, flowing phase was 0.3% phosphoric acid solution:acetonitrile= 94: 6 （V/V）; column temperature was 40℃ and sample size was 5 μL.2. Serum MHD concentration was detected using the constructed UPLC-UV method.6-methoxy salicylic acid was treated as the internal standard, and plasma sample was injected for analysis after protein precipitation with perchloric acid. Detection conditions were shown as follows: chromatographic column: 50 mm×2.1 mm ACQUITY UPLCBEH C18 1.7 μm chromatographic column （Waters, USA）;ACQUITY UPLC column on-line filtration system （Waters, USA）; the detection wave length was 254nm; flow velocity was 0.20 mL·min-1, flowing phase was 0.05%formic acid solution: acetonitrile= 77: 23 （V/V）; column temperature was 40℃,sample temperature was 4℃ and sample size was 5 μL.3. Basic clinical information from children confirmed with epilepsy and had received serum LEV concentration detection in outpatient or wards of Affiliated Children’s Hospital of Kunming Medical University from May 2016 to January 2017 was collected. SPSS 19.0 software was adopted for statistical analysis.4. Basic clinical information from children confirmed with epilepsy and had received serum MHD concentration detection in outpatient or wards of Affiliated Children’s Hospital of Kunming Medical University from May 2016 to January 2017 was collected. SPSS 19.0 software was adopted for statistical analysis.Results:1. Serum LEV concentration showed excellent linear relation within the range of 1.25-50 μg·mL-1,which obeyed the followed equation y = 0.0205x - 0.0149（R2=0.9996, r = 0.9999）. Such equation was associated with excellent specificity, with precision, accuracy, recovery rate and stability being within the limit requirements.2. Serum MHD concentration showed excellent linear relation within the range of 2.5-50 μg·mL-1, which obeyed the followed equation y = 0.1102x - 0.0058 （R2=0.9998, r = 0.9999）. Such equation was associated with excellent specificity, with precision, accuracy, recovery rate and stability being within the limit requirements.3. Differences in serum LEV concentration among different sex, age and weight were not statistically significant upon statistical analysis; difference in serum concentration between response group and non-response group was not statistically significant;difference in response rate between serum concentration of <4.17μg·mL-1 group and that of 4.17～12.48μg·mL-1 group was statistically significant; and differences in ALT,AST, TBIL, TP, Cr and UREA between affected children taking LEV group and healthy control group were not statistically significant.4. Differences in serum LEV concentration among different sex, age and weight were not statistically significant upon statistical analysis; difference in serum concentration between response group and non-response group was not statistically significant;difference in response rate among all serum concentration groups was not statistically significant. and differences in ALT, AST, TBIL, TP, Cr and UREA between affected children taking LEV group and healthy control group were not statistically significant.Conclusions:1. An UPLC method is constructed in this paper to determine serum LEV concentration, which can be applied in determining plasma LEV concentration in clinic, thus supporting the pharmacokinetic study of LEV as well as its related clinical follow-up study.2. An UPLC method .is constructed in this paper to determine plasma 10,11-dihydro-10-hydroxyl carbamazepine concentration, which is associated with the advantages of easily accessible internal standard, simple operation, good stability,easy. repetition and cost-effectiveness. As a result, it can be applied in determining plasma 10,11-dihydro-10-hydroxyl carbamazepine concentration in clinic.3. LEV treatment can lead to excellent efficacy and high response rate; in addition,serum LEV concentration is partly related to response rate,but it has no obvious injury to liver and kidney functions. Best efficacy can be achieved through combining serum concentration monitoring with clinical response rate of affected children to adjust dose. The blood concentration range of LEV in children who suffer from epilepsy in Kunming is 2.5～28.39 μg mL-1.4. OXC treatment can lead to excellent efficacy and high response rate; in addition,serum MDH concentration is partly related to response rate,but it has no obvious injury to liver and kidney functions. The blood concentration range of LEV in children who suffer from epilepsy in Kunming is 2.43～40.94 μg ·mL-1. |
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