Human cerebrospinal fluid serum

Fig. 11.8.4. HPLC of thiamine in human serum (a) and human cerebrospinal fluid (b). Chromatographic conditions stationary phase, pBondapak C18 (10 pm) reversed phase (300 X 3.9 mm I.D.) mobile phase, methanol-aqueous sodium citrate, pH 4.0, 0.05 mol/1 (45/55, v/v), sodium 1-octanesulphonate 10 mmol/1 temperature, ambient flow rate, 1.2 ml/min detection, post-column fluorescence (excitation at 367 nm, emission at 435 nm). Peaks 1, saUcylamide 2, thiamine. Reproduced from Wielders and Mink (1983), with permission.

Blood from normal subjects contains 14-60 m ig/ml, serum 25-75 mpg/ml, urine 100-200 m[ig of B8/ml of fluid cerebrospinal fluid contains less than 1 m ig/ml. Normal human liver, obtained by biopsy, contains between 10-20 mpg of B6/mg of dried tissue rat brain between 3.5-5 m(ig/mg. (cf. Table 9). 

Bradbury, M.W.B., et al. 1963. The distribution of potassium, sodium, chloride and urea between lumbar cerebrospinal fluid and blood serum in human subjects. Clin Sci 25 97. 

Special applications of CIEF have focused on studies of glycoproteins, antibodies, and proteins in serum (e.g., hemoglobin variants [30,37,39,77-90], transferrin forms [21,31,36,80,82,90,91]), and cerebrospinal fluid [92], The gly-coforms of recombinant human tissue-type plasminogen activator [93-95] and... 

Tucker RM, Williams PL, Arathoon EG, Levine BE, Hartstein Al, Hanson LH, Stevens DA. Pharmacokinetics of fluconazole in cerebrospinal fluid and serum in human coccidioidal meningitis. Antimicrob Agents Chemother 1988 32(3) 369-73. 

In most applications, the bioanalysis involves the analysis of a number of dmgs, or one dmg and (some of) its metabolites in biological fluids, especially whole blood, plasma, serum, or urine. However, other matrices are studied as well various tissues (skin, liver, brain, thyroid gland), faeces, hair, tear fluid, cerebrospinal fluid, semen. In most studies, the analysis of samples from human origin or from rats is performed, although the analysis of samples from rabbits, mice, minipigs, dogs, and monkeys is also performed. 

The determination of ddl [44] and d4T [45] in human serum was reported by online SPE-LC-MS with positive-ion ESI-MS. 3TC and ddl were applied as IS. The analysis time was less than 5 min for both methods. The LOQ was 10 ng/ml for both compounds. Another group described the determination of ddl and d4T in human plasma, bronchoalveolar lavage fluid (BALE), alveolar cells, PBMC, seminal plasma, cerebrospinal fluid (CSF), and tonsil tissue [57]. Depending on the matrix, either isocratic or gradient LC was applied after SPE sample pretreatment. Positive-ion ESI-MS in SRM mode was applied. The LOQ for both compounds were 2.0 ng/ml in plasma, 0.5 ng/ml in CSF, 0.4 ng/ml in alveolar cells, BALF, and PBMC, 1 ng/ml in seminal plasma, and 0.01 ng/mg in tonsil tissue. 

As discussed earlier in this section, OSHA has mandated that all U.S. laboratories have an exposure control plan. In addition, the National Institute for Occupational Safety and Health (NIOSH), a functional unit of the GDC, has prepared and widely distributed a document entitled Universal Pre-cautions that specifies how U.S. clinical laboratories handle infectious agents. In general it mandates that clinical laboratories treat aU human blood and other potentially infectious materials as if they were known to contain infectious agents, such as HBV, HIV, and other blood-borne pathogens. These requirements apply to all specimens of blood, serum, plasma, blood products, vaginal secretions, semen, cerebrospinal fluid, synovial fluid, and concentrated HBV or HIV viruses. In addition, any specimen of any type that contains visible traces of blood should be bandied using tliese Universal Precautions. 

Harling-Beig, C. J., Knopf, P. M., Merriam, J., and Cserr, H. F., Role of cervical lymph nodes in the systemic humoral immune response to human serum albumin microinfused into rat cerebrospinal fluid, J. Neumimmunol., 25, 185, 1989. 

Palfrreyman, J. W., Thomas, D. G. T., and Ratcliffe, J. G., Radioimmunoassay of human myelin basic protein in tissue extracts, cerebrospinal fluid and serum and its clinical application to patients with head injury, Chin. Chim. Acta, 82, 259, 1978. 

The method employing chlorophosphonazo(IIl) was applied to determine Ca in water, food and pharmaceuticals [1]. The detection limit was 0.03 mg f Ca. No interferences from elements, excess vitamins, amino acids or citrate and acetate ions were observed. The speetrophotometric method based on amino G acid chlorophosphonazo was proposed to determine Ca in human serum and cerebrospinal fluids [2). The reagent reacts with Ca with the formation of greenish blue complex (A ax at 670 nm = 7.2 x 10 1 mor em ). Beer s law is obeyed over the range 0.02-0.8 g ml Ca. No interferences were observed from the commonly coexisting species present in the examined materials. 

It is also unclear how serum ketone levels correlate with brain ketone levels. Cerebrospinal fluid ketone levels are elevated in animals (Appleton DeVivo, 1974) and in humans fasting or on the KD (Owen et al., 1967 DeVivo, 1983), but to a lesser degree than in serum. It will be important to learn whether regional, local, or even synaptic ketone concentrations are important for an anticonvulsant effect, especially with regard to whether ketones directly alter neuronal excitability. 

Physiological free copper concentrations are certainly decisive for determining a role for copper in PrP cellular function. It is important to recall the properties of Cu2+ in buffer solution. The solubility product (Ksp) of Cu(OH)2 at physiological pH is in the order of 10 20. i.e., rather small. It is therefore likely that it is bound to other compounds like, e.g., human serum albumin which binds copper in the picomolar range [92]. To evaluate whether PrP is a functional copper binding protein one has to determine the relative concentration of membrane attached PrPc and that of albumin in the cerebrospinal fluid which is about 3 pM [93]. However, the free concentration of copper has not been determined. Inside cells... 

Ritchie, S., Comprehensive metabolomic profiling of serum and cerebrospinal fluid Understanding disease, human variability, and toxicity, in Surrogate Tissue Analysis, 1st edition, Burczynski, M.E., and Rockett, J.C., Eds., CRC Press LLC, Boca Raton, FL, 2006, p. 165. 

It is clear from these data that for AEA the relative regional abundance in the brain does not correlate with the distribution of CBIR. AEA levels in the brain are equivalent to those of other neurotransmitters such as dopamine and serotonin, but at least 10-fold lower than the levels reported for GABA and glutamate. AEA has also been found in peripheral tissues such as human and rat spleen, which expresses high levels of CB2R. Small amounts of AEA were also detected in human serum, plasma, and cerebrospinal fluid (Felder et al., 1996). 

Numerous applications are now encountered where FA chromatographic profiles of a human physiological fluid or tissue are correlated to certain pathological conditions. A few representative examples will now be mentioned that include both free (non-esterified) FA and the saponified lipids. The identification of a methyl-branched FA (phytanic acid) in plasma of the patients with Refsum s disease [360] is now a widely known example of the power of GC in studying various metabolic defects. The profiles of FA from brain ti.ssue lipids were investigated for various neurological disorders [361,362] and in experimental animals [363]. Tichy et al. [364] determined FA in different lipids isolated from the cerebrospinal fluid while the FA profiles in cerebrospinal fluid differ from those in blood serum, no obvious correlations between the FA composition and human neurological complications were established at this time. 

The present Interest in the characterization of both animal and human secretions has paralleled the development in psychophysical measurement techniques and in analytical methods such as headspace concentration, gas chromatography, and the combination of gas chromatography/mass spectrometry (GC/MS) which have made it possible to routinely separate and identify submicrogram quantities of organic compounds. GC/MS profiling of the small organic compounds present in body secretions, such as blood serum, cerebrospinal fluid, and urine of diseased and healthy individuals, has provided useful diagnostic information (2). 

The identification of low abundance proteins present in complex biological samples (for example, human serum or cerebrospinal fluid) can be improved also by removing... 

Watkins J. C. and Evans R. H (1981) Excitatory ammo acid transmitters Ann Rev Pharmacol Toxicol 21, 165-204 Wood J. H, Hare T. A, Galeser B. S, Ballenger C, and Post R M (1979) Low cerebrospinal fluid 7-aminobutyric acid content in seizure patients Neurology 29, 1203-1208 Wurtman, R, J, Hefti, F, and Melamed E. (1981) Precursor control of neurotransmitter synthesis. Pharmacol Rev 32, 315-335 Yamamoto S, Kiyama S., Watanabe Y, and Makita M (1982) Practical gas-liquid chromatographic method for the determination of amino acids m human serum. / Chromatogr 233, 39-50... 

---