Urine HPLC chromatogram

Figure 4.4 HPLC chromatogram obtained on a 50 xl injection of human urine collected from 0-4 h after a 1000 mg dose of BW935U83. The locations for BW935U83 (4), its glucuronide conjugate (5), and 3-fluoro-ribolactone (6) are indicated...

Figure 4. HPLC chromatogram of 1 mL of urine using a reverse phase column...

Figure 8. A p -nonapetide, which has been shown to be capable of mimicking an amphiphilic a-peptidic helical structure in a pepide-protein interaction [26], was C-labeIled and injected into rats. After 24 hours (in serum, not shown) and after 4 days (in urine and in feces the minor and major secretion pathways, respectively), there was hardly any metabolism, see the HPLC chromatograms with radiodetection (DSPOl is the compound, the formula of which is shown above) [68],...

Fig. 2 Typical HPLC chromatograms of histidine. (A) Standard histidine and glutathione. Injected amounts histidine (His), 5 pmol glutathione (GSH), 500 pmol. (B) Human serum. (C) Human blood. (D) Human urine. See the section HPLC system and Conditions for chromatographic conditions.

Figure 32-4 Representative HPLC chromatograms for (a) working standard b, norma) feces c, normal urine d, feces—hereditary coproporphyria e, urine—congenita erythropoietic porphyria f, feces—variegata porphyria g, urine—porphyria cutanea tarda and h, feces—porphyria cutanea tarda chromatographic conditions as described in the appendix on the Evolve site that accompanies this book. Peaks are I, uroporphyrin- 2, uroporphyrin-l 3, heptacarboxyiate porphyrin-l 4, heptacarboxylate porphyrin-tl 5, hexacarboxylate porphyrin 6, pentacarboxylate porphyrin 7, coproporphyrin-1 8, coproporphyrin- ll 9, deuteroporphyrin-IX ...

Figure 2. HPLC Chromatograms of Ametryn Reference Standards and Rat Urine...

Figure 7. Comparison of HPLC Chromatograms of Goat Urine Samples Before and After 6 N HCl Hydrolysis...

The opioids and their metabolites can be identified very specifically in the acetonitrile gradients of HPLC/DAD. Even structurally similar compounds such as codeine and dihydrocodeine can be clearly differentiated, although, in analogy to GC/MS, it can be necessary here also to measure highly concentrated samples a second time after dilution. After the sample preparation, in contrast to GC/MS, no further steps are necessary. When heroin is consumed, not only do its metabolites appear in the HPLC chromatogram, but so also do the characteristic peak patterns of the variously metabolized substances noscapine and papaverine, whose presence in the urine increases the certainty of the analytical result. The sensitivity of detection is sufficient to resolve questions concerning results of immunological tests, so that this method can also be used as a universal method for the routine analysis of opioids. 

Fig. 7. HPLC chromatograms with UV detection of (A) SFUS spiked with standard (300.0 ng/mL) and I.S. (333.3 ng/mL) (B) a human urine sample added with I.S.

Figure 2 HPLC chromatogram of human urine sample (top) and authentic tetra-O-benzoyl glucose ureide (bottom).

The conditions for enzymatic analysis varied with the enzyme under study. In general, the reactions were started by the addition of the enzyme activity, and incubations were at 37°C. The reaction was stopped by the addition of 100% methanol, and precipitated protein was removed by centrifugation. Samples of the supernatant solution were analyzed by HPLC. A chromatogram of a glycosidase activity is shown in Figure 9.74. The peak of free 4-MU is noted. The enzyme was obtained from urine samples. 

FIGURE 7.6 A sample containing 1 l,M of oxazepam and an unknown concentration of temazepam in dog urine was injected into the HPLC/CLND/MS system, (a) CLND chromatogram, (b) and (c) MRM chromatograms for oxazepam and temazepam. (Adapted from Deng, Y. et al., Rapid Commun. Mass Spectrom., 18, 1681, 2004. With permission.)... 

Traditionally, it has been established that urine samples should be diluted 1 4 with water prior to an LC-MS analysis. Plumb et al. [92,93] reported obtaining good results from LC-MS analysis for rat urine after a 1 4 dilution with water. Waybright et al. [94] investigated the applicability of this dilution method to human urine which is less concentrated than mouse or rat urine. He concluded that a 1 4 dilution of human urine is not optimum for the detection of lower abundance metabolites. Although, the chromatogram for neat untreated urine was more intense, injection of neat urine on an HPLC column without column washing between injections was not recommended because of contamination of the HPLC column. 

The determination of this important substance has for a long time been performed exclusively by chromatographic methods, as immunological tests are not available. Although HPLC/DAD can be used, GC/MS is much more sensitive. Methods with a limit of detection of 1 ng/ml have been described for the determination of chlormethiazole from serum [61]. Using the sample preparation methods described in Chapter 7, it is easily possible without derivatization to detect from urine the consumption of a chlormethiazole capsule (192 mg) even on the day after consumption. Figure 8-37 shows the chromatogram of a sample with such a low concentration. 

Fig. 19.6 HPLC analysis of the female-produced sex pheromone isolated from female conditioned seawater (Hardege, unpublished data). The chromatogram shows the HPLC analysis of female C. maenas urine at 2-day post-moult (black line), and synthetic pheromone, UDP (red line). HPLC conditions used were Phenomenex RP Fusion column (4.6 x 250 mm) mobile phase 0.2 M KH2PO4 buffer, pH 5.5 1 mL min-1. Synthetic pheromone UDP (for structure see insert) was at a concentration of 10-5 M. The unresolved shoulder peak in the female sample at 3.3 min represents both tautomeric forms of UDP...

Figure A16.1 HPLC separation of components in a mixture. The chromatograms are from Ying Liu et al.. Behavioral and Brain Functions 3 47 (2009) doi.1186/1744-9081-3-47. They show analysis of morning urine samples from a normal subject (top) and a severely depressed female patient (below). The analysis is looking for significant differences in the content of peptides in the urine. The samples in such analysis are injected into the HPLC column and as the samples passes down the column the components separate so that they emerge (left to right) as separate sharp peaks, which can be detected by their u.v. light absorption properties. The difference between the normal subject and the patient is striking.

Fig. 4. Gas chromatographic separation of AF-FC 2 (active fraction of estrous urine extract after flash chromatography and TLC), upper chromatogram AF-HPLC (active fraction of estrous urine extract after FC, TLC, and HPLC), lower chromatogram, ly sample. DB-1-30N column (J W)(0.25 ym 0.32 mm) 30M helium carrier gas temperature-programmed 5%C/minute, 35°C to 330°C retention time depicted in minutes. Solvent peak eluted prior to 4 minutes.

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