中国药物评价
中國藥物評價
중국약물평개
Chinese Journal of Drug Evaluation
2015年
2期
73-75
,共3页
L-亮氨酸叔丁酯%Marfey试剂%柱前衍生化%高效液相色谱法
L-亮氨痠叔丁酯%Marfey試劑%柱前衍生化%高效液相色譜法
L-량안산숙정지%Marfey시제%주전연생화%고효액상색보법
Key Words ] L-Leucine Tert-Butyl Ester Hydrochloride%Marfey′s reagent%Pre-column derivatization%High Performance Liquid Chromatography( HPLC)
目的：建立柱前衍生化-高效液相色谱法（ HPLC）测定L-亮氨酸叔丁酯盐酸盐中对映异构体的方法。方法：采用Nα-（2，4-二硝基-5-氟苯基）-L-丙氨酰胺（Marfey试剂）作为柱前衍生试剂，反应温度40℃，反应时间2.5 h，衍生试剂加入量为35μL的1％Marfey溶液；色谱柱：Alltech Apollo C18（150 mm ×4.6 mm，5μm），流动相A：水，流动相B：甲醇，线性梯度洗脱，检测波长：340 nm，柱温：30℃，流速：1.0 mL· min－1，进样量：10μL。结果：L-亮氨酸叔丁酯盐酸盐在1.15～23.2μg· mL－1范围内线性关系良好，r＝0．9995；测得检测限为0.4 ng，定量限为1.7 ng；平均回收率104.4％，相对标准偏差（ RSD）为3.4％；重复性实验中RSD为3.3％；此方法耐用性良好，L-亮氨酸叔丁酯盐酸盐衍生产物溶液体系稳定。结论：本方法灵敏、简便、专属性强，能对L-亮氨酸叔丁酯盐酸盐的对映异构体进行检测及定量分析。
目的：建立柱前衍生化-高效液相色譜法（ HPLC）測定L-亮氨痠叔丁酯鹽痠鹽中對映異構體的方法。方法：採用Nα-（2，4-二硝基-5-氟苯基）-L-丙氨酰胺（Marfey試劑）作為柱前衍生試劑，反應溫度40℃，反應時間2.5 h，衍生試劑加入量為35μL的1％Marfey溶液；色譜柱：Alltech Apollo C18（150 mm ×4.6 mm，5μm），流動相A：水，流動相B：甲醇，線性梯度洗脫，檢測波長：340 nm，柱溫：30℃，流速：1.0 mL· min－1，進樣量：10μL。結果：L-亮氨痠叔丁酯鹽痠鹽在1.15～23.2μg· mL－1範圍內線性關繫良好，r＝0．9995；測得檢測限為0.4 ng，定量限為1.7 ng；平均迴收率104.4％，相對標準偏差（ RSD）為3.4％；重複性實驗中RSD為3.3％；此方法耐用性良好，L-亮氨痠叔丁酯鹽痠鹽衍生產物溶液體繫穩定。結論：本方法靈敏、簡便、專屬性彊，能對L-亮氨痠叔丁酯鹽痠鹽的對映異構體進行檢測及定量分析。
목적：건립주전연생화-고효액상색보법（ HPLC）측정L-량안산숙정지염산염중대영이구체적방법。방법：채용Nα-（2，4-이초기-5-불분기）-L-병안선알（Marfey시제）작위주전연생시제，반응온도40℃，반응시간2.5 h，연생시제가입량위35μL적1％Marfey용액；색보주：Alltech Apollo C18（150 mm ×4.6 mm，5μm），류동상A：수，류동상B：갑순，선성제도세탈，검측파장：340 nm，주온：30℃，류속：1.0 mL· min－1，진양량：10μL。결과：L-량안산숙정지염산염재1.15～23.2μg· mL－1범위내선성관계량호，r＝0．9995；측득검측한위0.4 ng，정량한위1.7 ng；평균회수솔104.4％，상대표준편차（ RSD）위3.4％；중복성실험중RSD위3.3％；차방법내용성량호，L-량안산숙정지염산염연생산물용액체계은정。결론：본방법령민、간편、전속성강，능대L-량안산숙정지염산염적대영이구체진행검측급정량분석。
Objective:To establish a new RP-HPLC spectrometric method for enantioseparation of L-Leucine Tert-Butyl Ester Hydro-chloride.Methods:The separation of enantiomer was studied using Nα-(2,4-dinitro-5-fluorophenyl)-L-alaninamide (Marfey′s reagent) as the pre-column derivatizi ng reagent,the optimum condition of derivative reaction were 1%Marfey′s reagent 35μL for 2.5 h at 40℃. An Alltech Apollo C18 column (150 mm ×4.6 mm, 5 μm) with a gradient elution , the mobile phase A was water and mobile phase B was methanol.The UV detector was set at 340 nm, the column temperature was set at 30℃,the flow rate was set at 1.0 mL/min and the injecting volume was set at 10μL.Results:The method was linear in the range of 1.15~23.2 μg· mL-1(r=0.9995),the limit of de-tection was 0.4 ng, and the limit of quantitation was 1.7 ng.The average recovery rate was 104.4% and RSD was 3.4%.The RSD in the repeatability test was 3.3%.The ruggedness of the method was verified.The sample solvent was stable as well as the reaction system during detection.Conclusion:The results indicated the method showed the advantages of high sensitivity, convenience and good specifici-ty,and could be applied for determine the enantiomers of L-Leucine Tert-Butyl Ester Hydrochloride.
