TITLE

Fast LC separation of a myoglobin digest: a case study using monolithic and particulate RP 18 silica capillary columns

AUTHOR(S)
Rozenbrand, Johan; Van Bennekom, Wouter P.; Unger, Klaus K.; de Jong, Gerhardus J.
PUB. DATE
August 2006
SOURCE
Analytical & Bioanalytical Chemistry;Aug2006, Vol. 385 Issue 6, p1055
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
A method was developed for the fast separation of a myoglobin digest using a monolithic RP 18 silica capillary column of 100 μm I.D. The results were compared with those obtained with a particulate RP 18 silica capillary column of 100 μm I.D. at a flow-rate between 0.6 and 1.2 μl/min. The digest was analyzed at the monolithic column at a flow-rate up to 2.8 μl/min. This high flow-rate could not be applied to the particulate column due to the high back-pressure. When the starting composition of the gradient was changed from 0 to 20% and a gradient steepness of 16%/min was used, the analysis time was less than 4 min. A positive Mascot identification score of 115 was achieved for the MS–MS data. When a lower gradient steepness was employed, the chromatographic resolution and the peak capacity did not increase for most compounds. The intraday repeatability for the retention time of the monolithic column was better than 1.5% at 2.8 μl/min and even less than 0.5% using a flow-rate of 0.6 or 1.0 μl/min. For the particulate column, it was between 0.5 and 1.4% for a flow-rate of 0.6 μl/min, probably due to the high column back-pressure. The interday reproducibility for the retention time of the monolithic column was less than 0.9% using a flow-rate of 1.0 μl/min.
ACCESSION #
21396819

 

Related Articles

  • MYOGLOBIN. Schnell, Zoanne Burgess; Van Leeuwen, Anne M.; Kranpitz, Todd R. // Davis's Comprehensive Handbook of Laboratory & Diagnostic Tests ;Jan2003, p743 

    Presents a comprehensive guide to laboratory and diagnostic tests for myoglobin. Reference value; Indications; Nursing implications and procedure.

  • Stereoselective and driving-force-dependent photoinduced electron-transfer reactions of zinc myoglobin with optically active N, N'-dimethylcinchoninium and N, N'-dimethylcinchonidinium ions. Takashima, Hiroshi; Araki, Ayako; Takemoto, Keiko; Yoshikawa, Naokazu; Tsukahara, Keiichi // Journal of Biological Inorganic Chemistry;May2006, Vol. 11 Issue 3, p316 

    In order to understand the detailed mechanism of the stereoselective photoinduced electron-transfer (ET) reactions of zinc-substituted myoglobin (ZnMb) with optically active molecules by flash photolysis, we designed and prepared new optically active agents, such as N, N'-dimethylcinchoninium...

  • Determination of myoglobin concentration in blood-perfused tissue. Masuda, Kazumi; Truscott, Kent; Ping-Chang Lin; Kreutzer, Ulrike; Youngran Chung; Sriram, Renuka; Jue, Thomas; Lin, Ping-Chang; Chung, Youngran // European Journal of Applied Physiology;Sep2008, Vol. 104 Issue 1, p41 

    The standard method for determining the myoglobin (Mb) concentration in blood-perfused tissue often relies on a simple but clever differencing algorithm of the optical spectra, as proposed by Reynafarje. However, the underlying assumptions of the differencing algorithm do not always lead to an...

  • In vitro monitoring of clindamycin in human urine using flow injection chemiluminescence. Xiaodong Shao; Xiaofeng Xie; Zhenghua Song // Microchimica Acta;Mar2007, Vol. 157 Issue 3/4, p159 

    A sensitive chemiluminescence method for the determination of clindamycin is presented. The method is based on the inhibitory effect of clindamycin on the chemiluminescence reaction between luminol and myoglobin in a flow-injection system. The decrement in chemiluminescence intensity is linear...

  • Effect of non-enzymatic glycation on esterase activities of hemoglobin and myoglobin. Subhrojit Sen; Tania Bose; Anjana Roy; Abhay Chakraborti // Molecular & Cellular Biochemistry;Jul2007, Vol. 301 Issue 1/2, p251 

    Abstract  Heme proteins––hemoglobin and myoglobin possess esterase activities. Studies with purified hemoglobin from normal individuals and diabetic patients revealed that the esterase activity as measured from hydrolysis of p-nitrophenyl acetate (p-NPA) was higher in...

  • Tilting after Dutch windmills: probably no long-lived Davydov solitons in proteins. Austin, Robert H.; Aihua Xie; Dan Fu; Warren, Warren W.; Redlich, Britta; van der Meer, Lex // Journal of Biological Physics;Jan2009, Vol. 35 Issue 1, p91 

    We present a summary of picosecond pump-probe and photon echo experiments in the mid-IR at 6 �m on the protein myoglobin. The intriguing temperature dependence of the amide I band in Mb is rather similar to the temperature dependence of the amide I band of acetanilide, the molecule that...

  • Physiology: Myoglobin's new clothes. Cossins, Andrew; Berenbrink, Michael // Nature;7/24/2008, Vol. 454 Issue 7203, p416 

    The article focuses on the study conducted to test the physiological effects of myoglobin especially in the production of nitric oxide which was known to limit tissue damage caused by restricted blood flow. Significant findings were revealed from the paper by U. B. Hendgen-Cotta and colleagues...

  • Prognostic value of serum myoglobin in patients after cardiac surgery. Hofmann, Denis; Buettner, Marco; Rissner, Florian; Wahl, Manuela; Sakka, Samir G. // Journal of Anesthesia;2007, Vol. 21 Issue 3, p304 

    Serum myoglobin as a marker of myocardial damage and injury has been shown to be of prognostic value in patients with cardiovascular events. In this study, we analyzed the prognostic value of serum myoglobin in comparison to other parameters of muscle damage and renal function in patients after...

  • Two species of antarctic icefishes (genus Champsocephalus) share a common genetic lesion leading to the loss of myoglobin expression. Grove, Theresa J.; Hendrickson, Jamie W.; Sidell, Bruce D. // Polar Biology;Sep2004, Vol. 27 Issue 10, p579 

    Species of the suborder Notothenioidei dominate the fish fauna of coastal Antarctic waters. Members of one notothenioid family, Channichthyidae (Antarctic icefishes), are unique among all vertebrates in lacking the circulating oxygen-binding protein hemoglobin. Icefish species also do not...

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics