The new method allowed chromatographic separation and quantification of 25OHD2, 25OHD3, along with 25OHD3 epimer 3-epi-25OHD3 and isobars 1-α-hydroxyvitamin-D3 (1αOHD3), and 7-α-hydroxy-4-cholesten-3-one (7αC4). To demonstrate the practical usefulness of our method, blood samples were collected from 5 healthy male Caucasian volunteers age range 22 to 37 years and 25OHD2, 25OHD3 along with co-eluting epimers and analogues were quantified. It involved i) liquid-liquid extraction, ii) tandem columns (a high resolution ZORBAX C18 coupled to an ULTRON chiral, with guard column and inlet filter), iii) Stanozolol-D3 as internal standard, and iv) identification via ESI and monitoring of three fragmentation transitions. MethodsĪ positive ion electrospray ionisation (ESI) LC-MS/MS method was used in the Multiple Reaction Monitoring (MRM) mode for quantification. The aim of this study was to develop a novel LC-MS/MS method that can accurately identify and quantitate 25OHD3 and 25OHD2 through chromatographic separation of 25OHD from its epimers and isobars. However, among other interferents, this method may be compromised by overlapping peaks and identical masses of epimers and isobars, resulting in inaccuracies in circulating 25OHD measurements. Liquid chromatography tandem mass spectrometry (LC- MS/MS) has been shown to facilitate accurate separation and quantification of the major circulating metabolite 25-hydroxyvitamin-D3 (25OHD3) and 25-hydroxyvitamin-D2 (25OHD2) collectively termed as 25OHD. Recently, the accuracies of many commercially available immunoassays for Vitamin D have been questioned.
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Since it's not a FPS - you can't just shoot in a straight line or adjust to the shell drop just a little, shells arcs are really high. The game world is 3D, but you're only able to aim at 2 axis. but, somehow it makes you miss ship B regardless!!! this is what i find fascinating? Its like ship B has a forcefield around it that prevented the shells from landing where they would have landed had ship B not been in the way I've already described to you how it works before. Now lets say there happened to be ship B right there, you were actually aiming at the water line of ship B, but the computer cannot know that, it only knows you aimed poorly at ship A. the shells hit just a bit too low *where you aimed* and missed. lets say you are locked on to ship A and you aim at the water line of ship A but then you slipped a little bit and aimed too low. You seem to ignore the fact that computer knows what ship you are looking at, and incidentally it keeps you locked at whatever ship you are locked at. Erm, if you are locked to ship A, by definition, computer thinks you are aiming at the pixelset-hitbox-watchmacallit of ship A, since you are far off those pixels aiming at ship B, you will of course have to rely on dispersion to hit. |
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