西南国防医药
西南國防醫藥
서남국방의약
MEDICAL JOURNAL OF NATIONAL DEFENDING FORCES IN SOUTHWEST CHINA
2014年
6期
589-591
,共3页
高效液相色谱法%儿茶酚氧甲基转移酶%原儿茶酸%香草酸%肝微粒体
高效液相色譜法%兒茶酚氧甲基轉移酶%原兒茶痠%香草痠%肝微粒體
고효액상색보법%인다분양갑기전이매%원인다산%향초산%간미립체
HPLC%catechol-O-methyltransferase(COMT)%protocatechuic acid%vanillic acid%liver microsome
目的：建立测定大鼠离体肝微粒中儿茶酚氧甲基转移酶（ COMT）底物（原儿茶酸）和代谢产物（香草酸）含量的HPLC法。方法离体肝微粒体样品经酸化后，乙酸乙酯提取。以绿原酸作为内标，色谱柱为 Welchrom C18，流动相为甲醇：0.1％磷酸＝25：75，流速为1 ml/min，检测波长为260 nm、326 nm，柱温30℃，进样量20μl。结果原儿茶酸和香草酸分别在0.5~20μg/ml和0.375~15μg/ml范围内线性关系良好，r≥0.9995；原儿茶酸的日内精密度和日间精密度RSD≤13.2％，方法回收率为90.3％~115.4％，提取率≥94.9％；香草酸的日内精密度和日间精密度RSD≤8.5％，方法回收率为93.4％~103.1％，提取回收率≥88.5％。结论该方法灵敏、简便、准确，可用于大鼠肝微体中COMT活性的测定。
目的：建立測定大鼠離體肝微粒中兒茶酚氧甲基轉移酶（ COMT）底物（原兒茶痠）和代謝產物（香草痠）含量的HPLC法。方法離體肝微粒體樣品經痠化後，乙痠乙酯提取。以綠原痠作為內標，色譜柱為 Welchrom C18，流動相為甲醇：0.1％燐痠＝25：75，流速為1 ml/min，檢測波長為260 nm、326 nm，柱溫30℃，進樣量20μl。結果原兒茶痠和香草痠分彆在0.5~20μg/ml和0.375~15μg/ml範圍內線性關繫良好，r≥0.9995；原兒茶痠的日內精密度和日間精密度RSD≤13.2％，方法迴收率為90.3％~115.4％，提取率≥94.9％；香草痠的日內精密度和日間精密度RSD≤8.5％，方法迴收率為93.4％~103.1％，提取迴收率≥88.5％。結論該方法靈敏、簡便、準確，可用于大鼠肝微體中COMT活性的測定。
목적：건립측정대서리체간미립중인다분양갑기전이매（ COMT）저물（원인다산）화대사산물（향초산）함량적HPLC법。방법리체간미립체양품경산화후，을산을지제취。이록원산작위내표，색보주위 Welchrom C18，류동상위갑순：0.1％린산＝25：75，류속위1 ml/min，검측파장위260 nm、326 nm，주온30℃，진양량20μl。결과원인다산화향초산분별재0.5~20μg/ml화0.375~15μg/ml범위내선성관계량호，r≥0.9995；원인다산적일내정밀도화일간정밀도RSD≤13.2％，방법회수솔위90.3％~115.4％，제취솔≥94.9％；향초산적일내정밀도화일간정밀도RSD≤8.5％，방법회수솔위93.4％~103.1％，제취회수솔≥88.5％。결론해방법령민、간편、준학，가용우대서간미체중COMT활성적측정。
Objective To establish a HPLC method for the simultaneous determination of content of catechol-O-methyltransferase( COMT)substrate( protocatechuic acid)and product( vanillic acid)in isolated rat liver microsomes. Methods The isolated liver microsome samples were acidified by HCl and then extracted with ethyl acetate;chlorogenic acid was chosen as internal standards;the detection was performed on a Welchrom C18 column,with mobile phase of methanol:0. 1% phosphoric acid=25:75;the flow rate was 1 ml/min and the detection wavelength were set at 260 nm and 326 nm;the column temperature was 30 ℃,and injection volume was 20 μl. Results The linear ranges of protocatechuic acid and vanillic acid were 0. 5-20 μg/ml and 0. 375-15 μg/ml, respectively;the intra-and inter-assay precisions of protocatechuic acid was less than 13. 2%,the method recovery was between 90. 3%and 115. 4%,and the extraction recovery was greater than 94. 9%;while the intra-and inter-assay precisions of vanillic acid were less than 8. 5%,the method recovery was between 93. 4% and 103. 1%,and the extraction recovery was greater than 88. 5%. Conclusions This method is sensitive,simple and accurate and can be applied to detect COMT enzyme activity in isolated rat liver microsomes.
