中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2011年
9期
769-774
,共6页
李影%唐爱国%穆萨%肖乐东
李影%唐愛國%穆薩%肖樂東
리영%당애국%목살%초악동
肾功能不全,慢性%氨基酸类,芳香%色谱法,高压液相
腎功能不全,慢性%氨基痠類,芳香%色譜法,高壓液相
신공능불전,만성%안기산류,방향%색보법,고압액상
Renal insufficiency,chronic%Amino acids,aromatic%Chromatography,high pressure liquid
目的 建立同时测定血清AAA含量的HPLC-FLD法，探讨CRI患者血清AAA含量变化及其临床应用价值。方法血清标本来自于100名健康体检者和80例CRI患者。将CRI患者按2002年美国肾脏基金会(NKF)诊断分期标准进行分期：CKD 2期4例、CKD 3期12例、CKD 4期12例和CKD 5期52例；按CRI不同病因分组：慢性肾炎型32例、糖尿病型36例和高血压型12例。血清经高氯酸去蛋白后，离心取上清液测定，外标法定量。采用Megres C18色谱柱，流动相为乙腈：水（体积比为1∶9），流速为1.0 ml/min,荧光检测器在不同时间段设定特定波长对血清AAA进行测定。对健康对照组和CRI患者组血清中AAA总量、Tyr、Phe和Trp含量及不同分期和不同病因CRI患者血清Tyr、Phe和Trp含量进行比较，同时评价血清AAA总量诊断CRI的敏感度与特异度。结果Tyr、Phe和Trp线性范围分别为0.550～275.000、3.050 ～1 220.000和0.049～49.000 μmol/L，最低检测限分别为0.014、0.500和0.005 μmol/L,平均回收率分别为100.9％、101.3％和98.5％，日内精密度为2.32％～3.92％（平均为3.13％），日间精密度为3.18％ ～4.20％（平均为3.58％）。CRI患者组血清AAA总量、Tyr、Trp含量及Tyr/Phe比值分别为(135.74±12.23)、(52.27±8.25)、(21.49±4.25) μmoL/L和[0.87（0.68 ～1.05）]，低于健康对照组的(174.47±11.57)、（63.53±4.68）、(44.22±3.67) μmol/L和[0.97（0.94～1.00）]，差异均有统计学意义（t=- 14.709、4.452、22.100,U=266.000,P均＜0.05）。不同分期CRI患者Tyr、Phe和Trp含量差异无统计学意义；Tyr含量在慢性肾炎组、高血压组和糖尿病组间差异无统计学意义，Phe在慢性肾炎组与高血压组、慢性肾炎组与糖尿病组间差异有统计学意义（U= 114.00、395.00，P均＜0.05），Trp在慢性肾炎组与糖尿病组间差异有统计学意义(U=349.00,P＜0.05)。血清AAA总量诊断CRI的敏感度、特异度分别为90％ (72/80)和100％ (100/100)。结论HPLC-FLD法测定血清AAA简便、快速，敏感度高及特异度好，同时测定血清AAA含量对CRI患者的诊断和评价有一定价值。
目的 建立同時測定血清AAA含量的HPLC-FLD法，探討CRI患者血清AAA含量變化及其臨床應用價值。方法血清標本來自于100名健康體檢者和80例CRI患者。將CRI患者按2002年美國腎髒基金會(NKF)診斷分期標準進行分期：CKD 2期4例、CKD 3期12例、CKD 4期12例和CKD 5期52例；按CRI不同病因分組：慢性腎炎型32例、糖尿病型36例和高血壓型12例。血清經高氯痠去蛋白後，離心取上清液測定，外標法定量。採用Megres C18色譜柱，流動相為乙腈：水（體積比為1∶9），流速為1.0 ml/min,熒光檢測器在不同時間段設定特定波長對血清AAA進行測定。對健康對照組和CRI患者組血清中AAA總量、Tyr、Phe和Trp含量及不同分期和不同病因CRI患者血清Tyr、Phe和Trp含量進行比較，同時評價血清AAA總量診斷CRI的敏感度與特異度。結果Tyr、Phe和Trp線性範圍分彆為0.550～275.000、3.050 ～1 220.000和0.049～49.000 μmol/L，最低檢測限分彆為0.014、0.500和0.005 μmol/L,平均迴收率分彆為100.9％、101.3％和98.5％，日內精密度為2.32％～3.92％（平均為3.13％），日間精密度為3.18％ ～4.20％（平均為3.58％）。CRI患者組血清AAA總量、Tyr、Trp含量及Tyr/Phe比值分彆為(135.74±12.23)、(52.27±8.25)、(21.49±4.25) μmoL/L和[0.87（0.68 ～1.05）]，低于健康對照組的(174.47±11.57)、（63.53±4.68）、(44.22±3.67) μmol/L和[0.97（0.94～1.00）]，差異均有統計學意義（t=- 14.709、4.452、22.100,U=266.000,P均＜0.05）。不同分期CRI患者Tyr、Phe和Trp含量差異無統計學意義；Tyr含量在慢性腎炎組、高血壓組和糖尿病組間差異無統計學意義，Phe在慢性腎炎組與高血壓組、慢性腎炎組與糖尿病組間差異有統計學意義（U= 114.00、395.00，P均＜0.05），Trp在慢性腎炎組與糖尿病組間差異有統計學意義(U=349.00,P＜0.05)。血清AAA總量診斷CRI的敏感度、特異度分彆為90％ (72/80)和100％ (100/100)。結論HPLC-FLD法測定血清AAA簡便、快速，敏感度高及特異度好，同時測定血清AAA含量對CRI患者的診斷和評價有一定價值。
목적 건립동시측정혈청AAA함량적HPLC-FLD법，탐토CRI환자혈청AAA함량변화급기림상응용개치。방법혈청표본래자우100명건강체검자화80례CRI환자。장CRI환자안2002년미국신장기금회(NKF)진단분기표준진행분기：CKD 2기4례、CKD 3기12례、CKD 4기12례화CKD 5기52례；안CRI불동병인분조：만성신염형32례、당뇨병형36례화고혈압형12례。혈청경고록산거단백후，리심취상청액측정，외표법정량。채용Megres C18색보주，류동상위을정：수（체적비위1∶9），류속위1.0 ml/min,형광검측기재불동시간단설정특정파장대혈청AAA진행측정。대건강대조조화CRI환자조혈청중AAA총량、Tyr、Phe화Trp함량급불동분기화불동병인CRI환자혈청Tyr、Phe화Trp함량진행비교，동시평개혈청AAA총량진단CRI적민감도여특이도。결과Tyr、Phe화Trp선성범위분별위0.550～275.000、3.050 ～1 220.000화0.049～49.000 μmol/L，최저검측한분별위0.014、0.500화0.005 μmol/L,평균회수솔분별위100.9％、101.3％화98.5％，일내정밀도위2.32％～3.92％（평균위3.13％），일간정밀도위3.18％ ～4.20％（평균위3.58％）。CRI환자조혈청AAA총량、Tyr、Trp함량급Tyr/Phe비치분별위(135.74±12.23)、(52.27±8.25)、(21.49±4.25) μmoL/L화[0.87（0.68 ～1.05）]，저우건강대조조적(174.47±11.57)、（63.53±4.68）、(44.22±3.67) μmol/L화[0.97（0.94～1.00）]，차이균유통계학의의（t=- 14.709、4.452、22.100,U=266.000,P균＜0.05）。불동분기CRI환자Tyr、Phe화Trp함량차이무통계학의의；Tyr함량재만성신염조、고혈압조화당뇨병조간차이무통계학의의，Phe재만성신염조여고혈압조、만성신염조여당뇨병조간차이유통계학의의（U= 114.00、395.00，P균＜0.05），Trp재만성신염조여당뇨병조간차이유통계학의의(U=349.00,P＜0.05)。혈청AAA총량진단CRI적민감도、특이도분별위90％ (72/80)화100％ (100/100)。결론HPLC-FLD법측정혈청AAA간편、쾌속，민감도고급특이도호，동시측정혈청AAA함량대CRI환자적진단화평개유일정개치。
Objective A HPLC-FLD method was developed to determine the levels of serum AAA in CRI patients, and to study the variation of serum AAA in CRI patients and its clinical significances. MethodsSerum samples were collected from 100 health controls and 80 CRI patients. According to 2002 National Kidney Foundation (NKF) staging diagnosis, CRI patients included 4 of stage 2, 12 of stage 3, 12 of stage 4, 52 of stage 5. According to pathogenesis, CRI patients were also divided into 3 groups :chronic nephritis group ( n = 32), DM group ( n = 36), hypertension group ( n = 12 ).Serums were deproteinized by equal volume of 5％ (v/v) PCA and supernate were analyzed direcdy. External standard method was used as quantitative method. The analytical column was Megres C18. 10％ acetonitrile in water was used as mobile phase. Flow rote was 1.0 ml/min. The wavelengths of fluorescence excitation and emission were changed with specific time. The levels of Tyr, Phe and Trp in CRI groups, different CKD stages and different pathogenesis were compared with healthy control groups to evaluated the sensitivity and specificity of serum AAA for CRI diagnosis. Results The linear ranges of the method were 0. 550 -275.000, 3. 050 - 1220. 000 and 0. 049 -49. 000 pμmol/L for Tyr, Phe and Trp, respectively. The limit of detection (LOD) was 0.014 μmol/L for Tyr, 0.500μmol/L for Phe, and 0.005 μmol/L for Trp. The average recovery was 100. 9％, 101.3％ and 98. 5％ for Tyr, Phe and Trp, respectively. Intra-day CVwas 3. 18％ -4. 20％ ( mean was 3. 13％ )and inter-day CV was 3. 18％ -4. 20％ ( mean was 3. 58％ ). The concentration of serum AAA, Tyr and Trp and the ratio of Tyr/Phe in CRI patients were( 135.74 ±23.23 )μmol/L, (52.27 +8.25) μmol/L, (21.49 ±4.25) μmol/L and[0.87(0.68 - 1.05)]μmol/L. which were lower than that in healthy groups (t value was -14. 709, 4.452, 22. 100, U value was 266.000,respectively, P＜0. 05). The concentration of serum AAA, Tyr and Trp and the ratio of Tyr/Phe in healthy groups were ( 174. 47 ± 11.57 ) μmol/L, ( 63.53 ± 4. 68 ) μmol/L, (44. 22 ± 3. 67 ) μmol/L and[0. 97(0. 94 - 1.00)]μmoL/L. There were no statistically significant difference between the different stage of CRI. Compared with the concentration of Tyr, Phe and Trp among chronic nephritis group, DM group,hypertension group, the concentration of Tyr had no significant changes among these three kinds of diseases (P ＞ 0. 05 ). The concentration of Phe had significant changes between Chronic nephritis group and DM group, Chronic nephritis group and hypertension group ( U = 395.00, 114. 00, P ＜ 0. 05 ) ; the concentration of Trp haad significant changes between Chronic nephritis group and DM group ( U = 349.00, P ＜ 0. 05 ).The diagnostic sensitivity and specificity of serum AAA for CRI were 90％ (72/80) and 100. 0％ (100/100).Conclusions The method of high-performance liquid chromatography with fluorescence detection ( HPLC-FLD) is simple, rapid, sensitive and specific. Simultaneous determination of serum AAA was benefit to the diagnosis and evaluation of CRI patients.
