BODY AND TISSUE FLUIDS

Zhang et al. [49] determined penicillamine in urine by the coupled technique of chemiluminescence detection and liquid chromatography. The urine sample was adjusted to pH 2 with 2 M H2S04 and centrifuged at 3000 rpm for 5 min. A 12 mL 

Wakabayashi et al. [51] determined penicillamine in serum by HPLC. Serum (0.1 mL) was vortex-mixed for 30 s with 50 pL of 0.1% EDTA and 0.2 mL of 10% TCA. The solution was centrifuged at 1500 x g and filtered. A 5 pL portion was analyzed on a Shodex C18 column (15 cm x 4.6 mm i.d.), using a mobile phase of 19 1 methanolic 0.05 M phosphate buffer (pH 2.8) containing 1 mM sodium octylsulfate and 10 pM EDTA. Liver or kidney samples were similarly extracted, and the extracts were cleaned up on a Bond-Elut cartridge prior to HPLC analysis. Detection was effected with an Eicom WE-3G graphite electrode maintained at +0.9 V versus Ag/AgCl. The calibration graph was linear up to 500 ng, and the detection limits were 20 pg. For 1 pg of penicillamine added to serum, liver, or kidney, the respective relative standard deviations (n = 5) were 3.6, 5.1, and 4.4%. 

Rabenstein and Yamashita [52] determined penicillamine and its symmetrical and mixed disulfides by HPLC in biological fluids. Plasma and urine were deproteinized with trichloroacetic acid, and HPLC was performed on a column (25 cm x 4.6 mm) or Biophase ODS (5 pm) with a mobile phase comprising 0.1 M phosphate buffer (pH 3) and 0.34 mM Na octylsulfate at 1 mL/min. Detection was with a dual Hg-Au amalgam electrode versus a Ag-AgCl reference electrode. (z )-penicillamine and homocysteine were determined at the downstream electrode at +0.15 V, and homocystine, penicillamine-homocysteine, and penicillamine disulfides were first reduced 

Wronski [55] described a method to separate and resolve penicillamine from physiological fluids. To urine (100 mL) was added 60 g of (NH4)2S04, the solution was filtered, mixed with 2 g of Na2S03, and 1 mL of 0.1 M EDTA in 20% triethanolamine, and shaken with 5-20 mL of 0.01-0.06 M tributyltin hydroxide in octane for 5 min. A portion (5-8 mL) of the organic phase was shaken with 0.2 mL of HC1 in 20% glycerol solution. A portion of the aqueous phase (5-10 pL per cm of the strip width) was applied to cellulose gel, and electrophoretic separation was performed by the technique described previously. The thiol spots were visualized with o-hydroxymercuri benzoic acid-dithiofluorescein and densitometry with a 588-nm filter. 

Webb et al. [56] determined free penicillamine in the plasma of rheumatoid arthritis patients. Plasma ultrafiltrate was mixed with trichloroacetic acid and 4-aminobenzoic acid as internal standards, and HPLC mobile phase to determine total reduced penicillamine. Plasma was vortexed with trichloroacetic acid, the precipitated protein was removed after 15 min by centrifuging, and the supernatant solution was filtered and mixed with 4-aminobenzoic acid. In each instance, a 50-pL portion of solution was analyzed on a 25-cm column of Spherisorb-NH2 (5 pm) at 25 °C, with an electrochemical detector having dual porous graphite electrodes set at 

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Pharmacokinetic studies are designed to measure quantitatively the rate of uptake and metaboHsm of a material and determine the absorbed dose to determine the distribution of absorbed material and its metaboHtes among body fluids and tissues, and their rate of accumulation and efflux from the tissues and body fluids to determine the routes and relative rates of excretion of test material and metaboHtes and to determine the potential for binding to macromolecular and ceUular stmctures. 

The microbial assay is based on the growth of l ctobacillus casei in the natural (72) or modified form. The lactic acid formed is titrated or, preferably, the turbidity measured photometrically. In a more sensitive assay, l euconostoc mesenteroides is employed as the assay organism (73). It is 50 times more sensitive than T. casei for assaying riboflavin and its analogues (0.1 ng/mL vs 20 ng/mL for T. casei). A very useful method for measuring total riboflavin in body fluids and tissues is based on the riboflavin requirement of the proto2oan cHate Tetrahjmenapyriformis which is sensitive and specific for riboflavin. 

TXRF is an ideal tool for microanalysis [4.21]. The analytical merits are that TXRF has a broad range of linearity (lO -lO atoms cm ) and it is extremely surface-sensitive and matrix-independent. TXRF can be applied to a great variety of different organic and inorganic samples such as water, pure chemicals, oils, body fluids and tissues, suspended matters, etc., down to the picogram range. 

Body fluids and tissues Tantalum is a very stable passive metal and completely inert to body fluids and tissues. Bone and tissue do not recede from tantalum, which makes it attractive as an implant material for the human body" . 

P-Alanine, a metabolite of cysteine (Figure 34-9), is present in coenzyme A and as P-alanyl dipeptides, principally carnosine (see below). Mammalian tissues form P-alanine from cytosine (Figure 34-9), carnosine, and anserine (Figure 31-2). Mammalian tissues transami-nate P-alanine, forming malonate semialdehyde. Body fluid and tissue levels of P-alanine, taurine, and... 

PLC studies in the field of medicine cover the processes of separation, purification, and determination of xenobiotics or endogenous substances present in body fluids and tissues. Such research is carried out with the following aims ... 

The limit of determination [or limit of quantitation (LOQ)] is defined in Directive 96/46/EC as the lowest concentration tested at which an acceptable mean recovery (normally 70-110%) and acceptable relative standard deviation (normally <20%) are obtained. The specific requirements for LOQ in crops, food, feed, soil, drinking and surface water, air, body fluids, and tissues are described in Section 4. Because the abbreviation LOD usually means limit of detection rather than limit of determination, the authors prefer not to use this abbreviation here in order to avoid confusion, and LOQ is used throughout. According to Directive 96/46/EC no data with regard to the limit of detection must be given. 

In this section, the general requirements laid down in Directive 96/46/EC and in the guidance document SANCO/825/00 are discussed. Furthermore, specific requirements for the different matrices (food of plant and animal origin, soil, water, air, and body fluids and tissues) will be illustrated. 

Owing to the complexity of multi-residue methods for products of animal origin, it is not possible to outline a simple scheme however, readers should refer to methods described in two references for detailed guidance (Analytical Methods for Pesticides in Foodstuffs, Dutch method collection and European Norm EN 1528. ) There is no multi-method specifically designed for body fluids and tissues. The latter matrix can be partly covered by methods for products of animal origin. However, an approach published by Frenzel et al may be helpful (method principle whole blood is hemolyzed and then deproteinized. After extraction of the supernatant, the a.i. is determined by GC/MS. The LOQ is in the range 30-200 ag depending on the a.i.). 

The general sample set for method validation parameters is the same for all matrices under consideration (except body fluids and tissues, see Section 4.2.5) ... 

Analytical methods for the determination of residues in body fluids and tissues must be submitted only if the a.i. is classified as toxic or highly toxic. The method has to be validated only at the LOQ in general blood 0.05 mgL and tissues 0.1 mgkg (meat or liver, if not investigated under food of animal origin, see Section 4.2.1). 

The sample set must include two fortification levels appropriate to the proposed LOQ and likely residue levels or 10 times the LOQ, except for body fluids and tissues (considered in Section 5.2.3) where validation data at the LOQ are sufficient. Five determinations should be made at each fortification level. In general, mean... 

Containment of body fluids and tissues Protection from harmful external stimuli (barrier functions)... 

Secondary Hazards Aerosols (blood, body fluids from animals) Blood and body fluids Body tissue Body fluids and tissue (from animals). 

The specific and sensitive GLC determination of griseofulvin in body fluids and tissues such as skin, sweat, urine and plasma with electron capture detection has been used by several investigators (34,51,52). 

Identification and Determination in Body Fluids and Tissue Griseofulvin has usually been determined in body fluids... 

The recognition of their structure permits the determination of vitamins by the tools of analytical chemistry, but while such methods are widely used in industrial production, the minute quantities in body fluids and tissues limit the purely chemical approach to a few members of this group present in relatively high concentration, e.g., vitamin C (K5). Microchemical methods are in use for the determination of thiamine, riboflavin, and some of the fat-soluble vitamins, based on the most sensitive colorimetric and, in particular, fluorometric techniques. Vitamin D, on the other hand, is determined by animal assay. 

The establishment of quantitative methods for the determination of vitamins in body fluids and tissues by microbiological assay techniques should stimulate the search for the significance of vitamins in disease, not only in nutritional deficiency, but in the much wider field of all metabolic disturbances. Functional vitamin deficiencies are produced by malabsorption, by inhibitors of the vitamin function through products of the body, and particularly through drugs and other toxic substances. Vitamin deficiencies may be relative deficiencies whenever an individual s metabolism is deranged so as to require enhanced quantities of a given vitamin to cure or to counteract certain symptoms as, e.g., in Darier s disease (keratosis follicularis) (P2a). 

Where die connection between a vitamin and a disease is less transparent, a wide field remains open for the discovery of meaningful correlations between vitamin content of body fluids and tissues and physiologic or pathologic events. 

Josefsson M, Kronstrand R, Andersson J, Roman M. 2003. Evaluation of electrospray ionization liquid chromatography-tandem mass spectrometry for rational determination of a number of neuroleptics and their major metabolites in human body fluids and tissues. J Chromatogr B Analyt Technol Biomed Life Sci 789 151. 

Although we can measure the amount of chloroform in the air that you breathe out, and in blood, urine, and body tissues, we have no reliable test to determine how much chloroform you have been exposed to or whether you will experience any harmful health effects. The measurement of chloroform in body fluids and tissues may help to determine if you have come into contact with large amounts of chloroform. However, these tests are useful only a short time after you are exposed to chloroform because it leaves the body quickly. Because it is a breakdown product of other chemicals (chlorinated hydrocarbons), chloroform in your body might also indicate that you have come into contact with those other chemicals. Therefore, small amounts of chloroform in the body may indicate exposure to these other chemicals and may not indicate low chloroform levels in the environment. From blood tests to determine the amount of liver enzymes, we can tell whether the liver has been damaged, but we cannot tell whether the liver damage was caused by chloroform. 

Plasma protein binding of meropenem is approximately 2%. The volume of meropenem distribution is 15.7 to 26.68 L. Meropenem penetrates well into most body fluids and tissues, including cerebrospinal fluid, achieving concentrations... 

Sparfloxacin - Sparfloxacin is well absorbed following oral administration. Steady-state concentration was achieved on the first day by giving a loading dose that was double the daily dose. Oral absorption of sparfloxacin is unaffected by administration with milk or food, including high-fat meals. Sparfloxacin distributed well into the body. It penetrates well into body fluids and tissues. Sparfloxacin is metabolized by the liver. It is excreted in the feces (50%) and urine (50%). 

Absorption/Disthbution - When given orally, cycloserine is rapidly absorbed, reaching peak plasma concentrations in 4 to 8 hours. It is widely distributed throughout body fluids and tissues cerebrospinal fluid levels are similar to plasma. 

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