高效液相色谱-圆二色谱法测定Warfarin

智能文字提取功能测试中

高压液相色谱（HPLC）是定量估计微量化合物以及分离和纯化混合物的最有效方法。高效液相色谱法也被广泛用于食品和医药产品的分析。在这些领域中，样品通常表现出光学活性，从而能够分析对映体过量以及分离和纯化这些对映体。JASCO已使CD检测器（PMT和InGaAs）与HPLC（CD-4095）兼容，并使HPLC流动池与J-1500和J-1700 CD光谱仪兼容。这两种型号都具有高灵敏度、高分辨率和宽动态范围，检测精度高达200 nm。本申请说明表明，J-1500和J-1700 CD光谱仪可与JASCO HPLC系统（CD-4095）结合使用，以分离和获得手性物质的CD色谱图。Application Note Measurement of warfarin using circulardichroism coupled with HPLC2/3Application Note CDS Spect r oscop y Measurement of warfarin using circular dichroism coupled with HPLC High-p r ess u re l iq u id chroma t ograp h y (HPLC ) is t h e m o st effec ti ve m eth o d fo r th e qua n ti t a ti ve e s t i mati o n o f t r ac e le v e l s of co m po u n ds a nd f o r mixtur e se pa r a ti o n an d p uri f ication . HP L C is also widely u sed f o r th e a n a l ysis o f f ood an d m e d i c in al p ro d u c t s. In th e se fie l d s ， s a mpl e s of t e n ex h i bi t o pt i ca l a ct i vity， al l o w in g fo r t h e a naly sis of en a n ti ome ri c excess a n d the separa ti on a n d p ur i fication of th ese e n a n ti o mers . JA SCO h as made b oth CD detec t o r s (PMT and InG aA s)c o mp a tibl e fo r u se wi th HPLC (C D -4095)， as we l l a s an HP LC f l o w cel l fo r u se w ith the J-1500 a n d J -1700 CD sp ectr o m eters . Both mo d e l s p rovide h i g h s en sit i vity ，reso lu ti o n ， and a w i de dy n am i c range ， with h igh a c c u r acy detection down to 200 nm . In th i s ap p l i ca ti o n n ote ， the CD meas ur eme n t o f warfa rin w as ob t a i ned u s i ng a J -1500 s p ectropolarimeter in stati c m o de a n d coupl e d to an HPLC system (C D -4095) for in-li n e CD detection o f t he HPLC e lu e n t . Keywords J -1500， c ir c u l a r di c h r o i sm， J-1700， H PL C， CD-4095，me d icinal prod uc ts ， c h ir a l a n a ly sis， ph ar m a ce u tical JASCO J-1500-PAL h igh -throughput system View product information at w ww .j a s coi n c .com JASCO INC. 28600 Mary's Court ， Ea s t on， MD 21601 U S A Experimental Data acquisition interval 0.1nm Response time 2 sec Spectral bandwidth 1nm Scan speed 100 nm/min Accumulations 1 time Path length 1mm Wavelength range 210-400nm Sample 200 ug/mL warfarin CD chromatogram measurement conditions Data interval 1 sec Response time 1sec Spectral bandwidth 1nm Path length 10mm Sample concentration 200 pg/mL Injection volume 10 pL Wavelength range 220，263nm Flow rate 0.5 mL/min Mobile phase pH 2.0 aqueous phosphoric acid/acetonitrile (40/60) Column CHIRALCEL OD-RH (4.6 mm 1.D. x150 mm L， 5 um) Results Fi g u re 1 s h ow s th e C D a n d a b so rpti o n sp ectr a of R -(+)-wa r fa r i n a n d S -(-)-w arfa r i n . C D pe a k s are o b s e rv ed a t 220， 263，a n d 306 nm w hic h ca n b e u sed f or th e m easu r em e nt of the C D c hr o mato g ram. 28600 Mary's Cour t ， E asto n ， MD 21601US A F igu r e 2 shows th e CD a n d a b sorption c h romatogram s of R -(+)-wa r farin and S-(-)-w a r fa r i n at 220 an d 263 nm . The m e a su r em e n t wa s p e r fo r med u sing th e T i m e Co u rse Me a su remen t m o d e and w a s initi a ted b y the trigg e r s i g n a l from t h e ma n u al inj ect o r o r au t osamp l e r . T h e pea k re t en t io n fo r R-(+)-war f a r i n i s obser v ed at 7.61 m in u t es an d the pe a k r e t e n ti o n pr ov i d es h igh er sen si t i v i t y . The CD pe a k area de tect ed at 220 nm i s six tim es l arg e r th a n th e pe ak a t 263 n m a nd the ar ea of th e p eak d e tecte d at 220 nm i s do u ble th at a t 263 n m. T h e a r ea ra ti o o f R -(+)-wa r fa rin a n d S -(-)-w ar f ar in i s 1：1. Figure 2. The CD and absorption chromatogram of warfarin detected at 220 nm (green) and 263 nm (blue). Conclusion This app l ication note dem o n s tr ate s th at the J-1500 a n d J-1700 C D spectrometers can be u s ed in conju n ction w i t h the J ASCO H PL C sy s t e m (C D-4095) i n o r d er t o s ep ara t e an d o bt a in C D chro mat og r a ms o n c h ir a l sub st a nc es . JASCO INC. 28600 Mary's Cour t ， E asto n ， MD 21601US A