南昌大学学报(医学版)
南昌大學學報(醫學版)
남창대학학보(의학판)
ACTA ACADEMIAE MEDICINAE JIANGXI
2015年
2期
21-24
,共4页
黄伟%王素莲%李瑛%刘大仁%胡晓
黃偉%王素蓮%李瑛%劉大仁%鬍曉
황위%왕소련%리영%류대인%호효
人肝微粒体%藏花酸%HPLC
人肝微粒體%藏花痠%HPLC
인간미립체%장화산%HPLC
human liver microsomes%crocetin%HPLC
目的:建立一种测定人肝微粒体孵育体系中藏花酸浓度的 HPLC 内标法。方法在人肝微粒体温孵体系中加入藏花酸样品进行孵育反应,反应结束后用冰乙腈终止反应。经沉淀法处理样品后,采用 Diamonsil C18色谱柱(4.6 mm×150 mm,5μm)进行分离。流动相:甲醇-2%冰乙酸水溶液(7525,v/v);流速:1.0 mL·min-1;柱温:30℃;紫外检测波长:440 nm;内标:吲哚美辛。结果藏花酸经孵育反应后质量浓度测定方法的线性范围为0.125~8μg·mL-1,线性关系良好(R 2=0.9999)。藏花酸低、中、高质量浓度的提取回收率分别为:97.24%、96.58%、98.12%,其 RSD 均小于15%,批内、批间差异<15%,准确度为85%~115%。结论该 HPLC 法简便、准确,符合生物样品测定的要求,能快速、可靠地检测人肝微粒体中藏花酸的浓度。
目的:建立一種測定人肝微粒體孵育體繫中藏花痠濃度的 HPLC 內標法。方法在人肝微粒體溫孵體繫中加入藏花痠樣品進行孵育反應,反應結束後用冰乙腈終止反應。經沉澱法處理樣品後,採用 Diamonsil C18色譜柱(4.6 mm×150 mm,5μm)進行分離。流動相:甲醇-2%冰乙痠水溶液(7525,v/v);流速:1.0 mL·min-1;柱溫:30℃;紫外檢測波長:440 nm;內標:吲哚美辛。結果藏花痠經孵育反應後質量濃度測定方法的線性範圍為0.125~8μg·mL-1,線性關繫良好(R 2=0.9999)。藏花痠低、中、高質量濃度的提取迴收率分彆為:97.24%、96.58%、98.12%,其 RSD 均小于15%,批內、批間差異<15%,準確度為85%~115%。結論該 HPLC 法簡便、準確,符閤生物樣品測定的要求,能快速、可靠地檢測人肝微粒體中藏花痠的濃度。
목적:건립일충측정인간미립체부육체계중장화산농도적 HPLC 내표법。방법재인간미립체온부체계중가입장화산양품진행부육반응,반응결속후용빙을정종지반응。경침정법처리양품후,채용 Diamonsil C18색보주(4.6 mm×150 mm,5μm)진행분리。류동상:갑순-2%빙을산수용액(7525,v/v);류속:1.0 mL·min-1;주온:30℃;자외검측파장:440 nm;내표:신타미신。결과장화산경부육반응후질량농도측정방법적선성범위위0.125~8μg·mL-1,선성관계량호(R 2=0.9999)。장화산저、중、고질량농도적제취회수솔분별위:97.24%、96.58%、98.12%,기 RSD 균소우15%,비내、비간차이<15%,준학도위85%~115%。결론해 HPLC 법간편、준학,부합생물양품측정적요구,능쾌속、가고지검측인간미립체중장화산적농도。
Objective To establish an HPLC method for determining the concentration of cro-cetin in human liver microsomes.Methods After incubation with crocetin in the microsomal incu-bation system,the reaction was terminated and protein was precipitated by ice-cold acetonitrile. The HPLC analysis was performed with a reversed-phase Diamonsil C18 column (4.6×150 mm, 5 μm)at a column temperature of 30 ℃.The mobile phase consisted of methanol and 2% glacial acetic acid(7525,v/v)with a flow rate of 1.0 mL·min-1 .The detection wavelength was 440 nm.The indometacin was used as the internal standard.Results The linear range of this assay was 0.125-8 μg·mL-1 (R 2 =0.999 9).The extraction recoveries of low,medium and high concen-trations of crocetin were 97.24%,96.58% and 98.12%,respectively.The intra-and inter-batch relative standard deviation (RSD)was less than 15%,and the accuracy ranged from 85% to 115%.Conclusion The established HPLC method is simple,reliable and rapid for the detection of crocetin in human liver microsomes and meets the requirements for biological sample determina-tion.