中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2014年
4期
281-284
,共4页
谈绮文%郭玮%顾梅秀%张春燕%潘柏申
談綺文%郭瑋%顧梅秀%張春燕%潘柏申
담기문%곽위%고매수%장춘연%반백신
尿分析%草酸%柠檬酸%电泳,毛细管
尿分析%草痠%檸檬痠%電泳,毛細管
뇨분석%초산%저몽산%전영,모세관
Urinalysis%Oxalic acid%Citric acid%Electrophoresis,capillary
目的 建立以磷酸二氢钾为缓冲体系测定尿液中微量草酸和柠檬酸的毛细管电泳分析法,考察了缓冲液体系选择、缓冲液pH值、缓冲液浓度、分离电压、进样时间对分离测定的影响.方法 在毛细管区带电泳模式下,采用非涂层石英毛细管、磷酸二氢钾缓冲液进行电泳,直接紫外检测.收集2013年2月至3月间复旦大学附属中山医院尿石症患者24 h尿标本5份,健康体检患者24 h尿液标本5份,对该方法进行系统的方法学评价.对优化分离条件下检测的出峰时间与峰面积进行重复性验证,标准样品的浓度线性验证和加样回收率验证.结果 当毛细管柱内径为50 μm,长度为50 cm(有效长度为40 cm)、分离电压为-30 kV、缓冲溶液采用50 mmol/L(pH 6.5)、检测波长为200 nm时,尿液中草酸和柠檬酸在5 min内得到基线分离,草酸和柠檬酸迁移时间和峰面积的批内标准偏差分别小于1.0％和3.0％,批间标准偏差小于2.0％和4.0％,检测限均为1 mg/L,尿液中草酸和柠檬酸浓度在0～500 mg/L范围内线性关系均良好(r=0.999 5、0.995 4,P ＜0.05),回收率分别为102.38％和92.74％.结论 该方法简单、准确、成本低,可为尿液样品中微量草酸和柠檬酸的分析测定提供参考.
目的 建立以燐痠二氫鉀為緩遲體繫測定尿液中微量草痠和檸檬痠的毛細管電泳分析法,攷察瞭緩遲液體繫選擇、緩遲液pH值、緩遲液濃度、分離電壓、進樣時間對分離測定的影響.方法 在毛細管區帶電泳模式下,採用非塗層石英毛細管、燐痠二氫鉀緩遲液進行電泳,直接紫外檢測.收集2013年2月至3月間複旦大學附屬中山醫院尿石癥患者24 h尿標本5份,健康體檢患者24 h尿液標本5份,對該方法進行繫統的方法學評價.對優化分離條件下檢測的齣峰時間與峰麵積進行重複性驗證,標準樣品的濃度線性驗證和加樣迴收率驗證.結果 噹毛細管柱內徑為50 μm,長度為50 cm(有效長度為40 cm)、分離電壓為-30 kV、緩遲溶液採用50 mmol/L(pH 6.5)、檢測波長為200 nm時,尿液中草痠和檸檬痠在5 min內得到基線分離,草痠和檸檬痠遷移時間和峰麵積的批內標準偏差分彆小于1.0％和3.0％,批間標準偏差小于2.0％和4.0％,檢測限均為1 mg/L,尿液中草痠和檸檬痠濃度在0～500 mg/L範圍內線性關繫均良好(r=0.999 5、0.995 4,P ＜0.05),迴收率分彆為102.38％和92.74％.結論 該方法簡單、準確、成本低,可為尿液樣品中微量草痠和檸檬痠的分析測定提供參攷.
목적 건립이린산이경갑위완충체계측정뇨액중미량초산화저몽산적모세관전영분석법,고찰료완충액체계선택、완충액pH치、완충액농도、분리전압、진양시간대분리측정적영향.방법 재모세관구대전영모식하,채용비도층석영모세관、린산이경갑완충액진행전영,직접자외검측.수집2013년2월지3월간복단대학부속중산의원뇨석증환자24 h뇨표본5빈,건강체검환자24 h뇨액표본5빈,대해방법진행계통적방법학평개.대우화분리조건하검측적출봉시간여봉면적진행중복성험증,표준양품적농도선성험증화가양회수솔험증.결과 당모세관주내경위50 μm,장도위50 cm(유효장도위40 cm)、분리전압위-30 kV、완충용액채용50 mmol/L(pH 6.5)、검측파장위200 nm시,뇨액중초산화저몽산재5 min내득도기선분리,초산화저몽산천이시간화봉면적적비내표준편차분별소우1.0％화3.0％,비간표준편차소우2.0％화4.0％,검측한균위1 mg/L,뇨액중초산화저몽산농도재0～500 mg/L범위내선성관계균량호(r=0.999 5、0.995 4,P ＜0.05),회수솔분별위102.38％화92.74％.결론 해방법간단、준학、성본저,가위뇨액양품중미량초산화저몽산적분석측정제공삼고.
Objective To establish a method for determining oxalate and citrate in urine simultaneously by capillary electrophoresis.The components,the concentration and pH of the buffer solution,the separation voltage and the injection time on theseparation were studied in detail.Methods The separations were carried out using potassium dihydrogen phosphatebuffer ina fused-silica capillary tubeby capillary zone electrophoresis (CZE) and the detection were monitored by UV.24 h-urine samples from patients (n =5) and health control (n =5) were collected from Zhongshan Hospital of Fudan University for systematically validating the method developed.Results The optimized separations were carried out using a 50 mmol/L potassium dihydrogen phosphatebuffer solution (pH 6.5) in a fused-silica capillary tube of 50 cm × 50 μm I.D.Injections were made by using the pressure mode for 10 s at 34 mbar.The detections were monitored by a UV at 200 nm after samples were separated at avohage of 30 kV.Under the seconditions,urinary oxalate and citrate were separated completely within 5 min.The relative standard deviations of migration time and peak area within-run foroxalate and citrate were less than 1％ and 3.0％ and the betweenrun relative standard deviations were less than 2.0％ and 4.0％,respectively.The detection limits were 1 mg/L for both oxalate and citrate.The linearity ranges of oxalate and citrate were both 0-500 mg/L with the correlation coefficient between 0.999 5 and 0.995 4 (P ＜ 0.05),respectively.The average recoveries were 102.38％ for oxalate and 92.74％ for citrate.Conclusion This method is proved to be simple,sensitive and accurate,and also applied to determine oxalate and citrate in urine samples with satisfactory results.
