Chromatogram urine

Fig. 1 Chromatograms of urine samples containing THC metabolites detection with fast blue salt RR (A) and fast blue salt B (B). Track Ai and B, metabolite-free urines tracks A,6 and B15 represent ca. 60 ng total cannabinoids per ml urine (determined by RIA).

Figure 11.4 Chromatograms of plasma samples on a silica-chiralcel OJ coupled column system (a) plasma spiked with oxprenolol (internal standard) (b) plasma spiked with 040 p-g/ml metyrapone and 0.39 p-g/ml metyrapol (racemate) (c) plasma sample obtained after oral administration of 750 mg metaiypone. Peaks are as follows 1, metyrapone 2, metyrapol enantiomers 3, oxprenolol. Reprinted from Journal of Chromatography, 665, J. A. Chiarotto and I. W. Wainer, Determination of metyrapone and the enantiomers of its chfral metabolite metyrapol in human plasma and urine using coupled achfral-chfral liquid cltro-matography, pp. 147-154, copyright 1995, with permission from Elsevier Science.

It is appropriate at this juncture to illustrate the power of chemiluminescence in an analytical assay by comparing the limits of sensitivity of the fluorescence-based and the chemllumlnescence-based detection for analytes in a biological matrix. The quantitation of norepinephrine and dopamine in urine samples will serve as an illustrative example. Dopamine, norepinephrine, and 3,4-dihydroxybenzy-lamine (an internal standard) were derivatized with NDA/CN, and chemiluminescence was used to monitor the chromatography and determine a calibration curve (Figure 15). The limits of detection were determined to be less than 1 fmol injected. A typical chromatogram is shown in Figure 16. 

Note When this reagent sequence is combined with the in situ measurement of the UV spectra of the chromatogram zones before derivatization this reagent sequence becomes virtually specific for hydrochlorothiazide and its related derivatives in the analysis of urine. Hence, it is possible to use it for drug monitoring and doping control. 

Note The three reagents should be applied as quickly as possible after each other. In combination with the Ry value, and with UV detection before application of the reagent sequence this procedure allows the identification of therapeutic quantities of thiazide diuretics and methyldopa in urine together with a series of other therapeutic agents. Mobile phase residues e. g. acetic acid, should be completely removed from the chromatograms before application of the reagent sequence. 

Figure 32, Chromatograms of plasma and urine samples with various abnormalities, A, Phenylalaninemia B, tyrosinemia C, elevated plasma methionine seen in homocystinuria D, glycinemia E, normal urine F, argininosuccinic aciduria G, homocystinuria H, hyperglycinuria I, hyperlysinuria.

Figure 3. Typical chromatogram of UV-ahsorhing constituents of a 0.15-ml sample of urine. Conditions 150-cm anion exchange column (1215 fi, Aminex A ZI) bujfer, acetate at pH 4.4 gradient, buffer concentration from 0.015M to 6M flow rate, 10.5 ml/hr, coUimn temperature, 25°C for first 5 hr and 60°C thereafter (34).

Figure 8 Chromatograms of diluted urine (I) and diluted urine spiked with 2.1 pM choline and 2.3 pM acetylcholine (II). Elution order Hydrogen peroxide (0.7 min), choline (1.6 min), and acetyl choline (2.9 min). See Ref. 50 for further details.

TIC) for each urine sample. For comparison of TICs, Agilent ChemStation software allows one easily to stack (overlay) several chromatograms in the same computer window (Fig. 2.2). This feature helps to identify compounds with common retention times among several samples, but does not contain any mass spectral data for verification of a common identity for these peaks and is most useful for comparison of only a few TICs or a few compounds of interest. In these cases, mass spectral comparisons can be made quickly by visual inspection of the relevant spectra. 

In the course of studies on other pathological amino acidurias, the accompanying peptiduria has also been observed by many authors. Rapp de Eston et al. (R2) observed a marked increase in the excretion of both free amino acids and peptides in patients with diffuse hepatic necrosis. Using a simplified chromatographic method adapted to clinical purposes and suitable for analysis of amino acids excreted with urine, Skarzynski et al. (S5) demonstrated a raised level of a certain peptide which is always present in normal urine in smaller quantities, as well as the appearance of some new peptides in cases of jaundice and liver cirrhosis. Some abnormal peptide spots were also detected on the chromatograms in cases of progressive muscular dystrophy (K4) and in patients irradiated with X-rays (S2). 

Relative Proportions of Amino Acids in Bands Eluted from the Chromatograms OF THE NoNDIFFUSIBLE FRACTION OF URINE SEPARATED... 

Fig. 14.13. Chromatograms showing single and comprehensive two-dimensional GC of drugs extracted from urine. (A) GC-FID chromatogram, (B) Pulsed GcxGC chromatogram (C) GCxGC contour plot. Adapted with permission from Ref. [15, Fig. 14.3]. Complete details on separation conditions and analyte identity can be found in the original paper.

Fio. 60. Chromatogram of acidified urine extract. Column 5>/ m octadecyl silica, 25 cm X 4.6 mm i.d., temperature, 70 C flowrate, 2.0 ml/min. Gradient elution from 0.1 M phosphaM buffer, pH 2.1, with acetonitrile to about 4096 (v/v) organic solvent concentration. Sample size 10 containing the extract of 100 of urine. Reprinted with permission from Horvath et al. (3S4). 

Figure 8 shows typical chromatograms for spiked and unspiked rat urine. Recovery was found to be 50, 55, and 35% for three replicate samples spiked at the 2 yg/L level, which we have found to be typical recovery values for trace levels of DCB in aqueous media (e.g. industrial wastewater). 

Figure 9 shows a chromatogram for human urine spiked at 50 pg/L. Recovery was found to be 58% at this spike level. 

The chromatographic conditions were the same as for the determination of benzidine in hair dye formulations, except for the flow rate, which was 1 mL/min. Figure 10 shows typical chromatograms for spiked and unspiked urine. 

Fig. 3.1.7 Detection of HG by the GC-MS TIC method in the urine of patients with MCAD deficiency collected at different clinical statuses. A, left panel Organic acid profile of an acutely ill patient. The arrow indicates the portion of the chromatogram shown in the middle panel. Peak labeling 1 HG, 2 4-hydroxyphenylacetic acid. Right panel extracted ion chromatograms of the [M-15]+ ion of HG (m/z 230 red) and 4-hydroxyphenylacetic acid (m/z 281). Patient recovering from an acute episode. C Asymptomatic patient. The latter profile represents a situation where there is a high probability that HG may not be detected by a GC-MS TIC method...

Fig. 4.1.8 Thin-layer chromatography of patient urines and a normal control. The MPS types are indicated beneath each band. The positions of dermatan sulfate (DS), heparan sulfate (HS), chondroitin sulfate (CS), and keratan sulfate (KS) are given on the right. The chromatogram was kindly provided by Professor E. Paschke, Laboratory of Metabolic Disease, Department of Pediatrics, University of Graz, Austria...

Fig. 4.7.2 Multiple reaction monitoring (MRM) chromatogram of pooled urine spiked with C7-polyols produced by method 1 (without separation of polyol isomers). MRM transitions are given for each mass transition. IS Internal standard...

Fig. 4.7.3 MRM chromatogram of pooled urine spiked with C7-polyols produced...

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