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Biogenic Amine Assays for Pharmaceutical and Specialty Research

-summary product data: please request full insert for current and more detailed information

The following Elisa kits are manufactured by IBL (Immuno Biological Laboratories, Hamburg Germany) and distributed by RDI Divison of Fitzgerald Industries Intl for in vitro research use only-not for use in or on humans or animals, not for use in diagnostics.

Serotonin ELISA  cat#RDI-RE59121

 Enzyme Immunoassay for the Quantitative Determination of Serotonin  in Plasma, Urine, Tissue homogenates and Cell Culture Supernatants.

Price: $594 - 1 Kit Catalogue No : RE 59121 Product group : Tumor Markers 12 x 8 Product name : Serotonin ELISA 96 Method : ELISA Incubation time: over night, 2h, 1h Standard curve : 0.05 ng/ml - 11 ng/ml Sample/Prep. : 20 µl of Serum (urine, platelet-extract, tissue homogenat) Isotope/Substr.: PNPP, 405nm
CONTENTS Introduction Contents of the Kit Principle of the Test Test Procedure Performance Characteristics Expected Values Alternative Applications (More) Clinical Background Sales Arguments Product Literature Miscellaneous


Back to Contents Serotonin is an intermediate product of tryptophan metabolism and is located primarily in the enterochromaffin cells of the intestine, serotonergic neurons of the brain, platelets of the blood and is well established as a neurotransmitter in the central nervous system. Central serotonergic neurons have been associated with physiological functions such as sleep, endocrine regulation, and cardiovascular control . Nearly all of the serotonin in circulating blood is concentrated in platelets. Altered concentrations of circulating serotonin have been implicated in several pathologic conditions including chronic tension headache, schizophrenia, hypertension, Huntington's disease, Duchenne's muscular dystrophy and early acute appendicitis. The determination of serum serotonin levels is of high clinical significance for diagnostic assessment of carcinoid syndrome. An increasing interest in the determination of serotonin in platelets including uptake and release kinetics is expected in the near future.

Contents of the Kit

Back to Contents A.) SAMPLE PREPARATION 1. Assay Buffer 1 bottle 50 ml, concentrate, phosphate buffer with BSA and stabilizer, dilute 1:10 with bidist. water 2. Acylation Reagent 1 (3%) 1 vial 2 ml, ready for use, acetic acid anhydride in acetone. 3. Acylation Reagent 2 (10%) 1 vial 2 ml, ready for use, acetic acid anhydride in acetone B). ENZYME IMMUNOASSAY 1. Microtiter strips 12 break apart strips each 8 wells, coated with anti-rabbit IgG from goat 2. Serotonin-Biotin 1 vial 6 ml, ready for use, yellow coloured. 3. Serotonin Antiserum 1 vial 6 ml, ready for use, antiserum from rabbit with stabilizers, blue coloured. 4. Enzymconjugate, Concentrate 1 vial 200 µl, concentrate, anti-Biotin antibody from goat conjugated to alkaline phosphatase in Tris buffer with stabilizers, dilute 1:100 with Assay Buffer 5. Standards A-G 7 vials 0.5 ml each, raedy for use, contain acylated serotonin with stabilizers Concentrations (ng/ml / nmol/l) : A : 0 / 0 B : 0.05 / 0.26 C : 0.14 / 0.80 D : 0.40 / 2.30 E : 1.20 / 7.00 F : 3.70 / 21 G : 11 / 62.5 6. Control Serum 1 and 2 2 vials lyophilized, dissolve in 0.5 ml bidist. water each, for serotonin concentration see quality control certificate. Store dissolved controls at -20° Celsius or lower. 7. Wash Buffer 1 bottle 50 ml, concentrate, phophate buffer with Tween and stabilizer, dilute 1:20 with bidist. water 8. PNPP Substrate Tablets 9 Tablets contain p-nitrophenyl phosphate (PNPP) 9. PNPP Substrate Buffer 1 vial 30 ml, raedy for use, contains diethanolamine 10. PNPP Stop Solution 1 vial 10 ml, ready for use, contains 1N NaOH with 0.25 M EDTA 11. Adhesive Foil 3 pieces

Principle of the Test

Back to Contents The Serotonin-ELISA kit provides materials for the quantitative measurement of chemically derivatized serotonin in serum, plasma, urine, tissue homogenates and tissue culture supernates. The sample preparation, i.e. derivatization of serotonin to N-acylserotonin, is part of the sample dilution and is achieved by incubation of the respective sample with the "Acylation Reagent". The assay procedure follows the basic principle of competitive ELISA whereby there is competition between a biotinylated and a non-biotinylated antigen for a fixed number of antibody binding sites. The amount of biotinylated antigen bound to the antibody is inversely proportional to the analyte concentration of the sample. When the system is in equilibrium, the free biotinylated antigen is removed by a washing step and the antibody bound biotinylated antigen is determined by use of anti-biotin alkaline phosphatase as marker and p-nitrophenyl phosphate as substrate. Quantification of unknowns is achieved by comparing the enzymatic activity of unknowns with a response curve prepared by using known standards.

Test Procedure

Back to Contents 1. Summary 1. Pipette 50 µl of each Standard, acylated Control Serum and acylated patient samples. 2. Pipette 50 µl of Serotonin-Biotin. 3. Pipette 50 µl of Antiserum. 4. Incubate 16-20 h (over night) at 2-8° Celsius. 5. Wash each well three times with Wash Buffer. 6. Pipette 150 µl of Anti Biotin AP. 7. Incubate for 120 minutes at room temperature. 8. Wash each well three times with Wash Buffer. 9. Pipette 200 µl PNPP Substrate Solution. 10. Incubate at room temperature for 60 minutes. 11. Stop the substrate reaction by adding 50 µl of PNPP Stop Solution. 12. Read the optical density at 405 nm (reference wave length 600-650 nm) with a microtiterplate reader within 60 minutes. 2. Detailed Instructions for Use Specimen Collection and Storage Serum The useful precautions for venipuncture should be observed. Blood has to be centrifuged within one hour after collection. The serum obtained should be applied in the assay after storing for up to two hours at room temperature or for up to 8 hours at 4 - 8 °C. If stored for longer periods the serum should be frozen at - 20 °C or lower. Repeated thawing and freezing should be avoided. Urine The total volume of urine excreted during a 24 hour period should be collected and mixed in a single bottle containing 10 -15 ml of 6N hydrochloric acid as preservative. The exposure of samples to direct sun light should be avoided. Urine samples which are not assayed immediatly may be stored at - 20 °C or lower for at least 6 months. Plasma samples and platelets More than 98 % of the circulating serotonin is located in th platelets and is released during blood clotting. Blood must be collected by venipuncture into plastic tubes containing EDTA or Citrate as anticoagulant (e.g. 10 ml Monovette NC with 1 ml citrate from SARSTEDT). Samples are kept and centrifuged at room temperature for 10 min at 200 x g to obtain platelet-rich plasma (PRP). The PRP-supernatant is then transferred to another tube and the platelets counted. To obtain the platelet pellet, an aliquot of 100 µl of PRP (containing between 350.000 and 500.000 platelets/µl) is added to 900 µl of physiological saline and centrifuged at 4500 x g for 10 min at 4 °C (or at 10000 x g for 2 min at 4 °C). The supernatant is then discarded. 200 µl of bidistilled water is added to the pellet, which can then be stored frozen at < - 20 °C for several weeks without any alteration of serotonin content. After thawing of the frozen samples centrifuge at 10000 x g for 2 min at room temperature. 20 µl of the supernatant are used in the ELISA (see Preparation of samples). If you want to measure serotonin in platelet-free plasma (PFP), an aliquot of the PRP is centrifuged at 4500 x g for 10 min at 4 °C (or at 10000 x g for 2 min at 4 °C) to obtain PFP. 50 µl of the supernatant are used in the ELISA for the measurement of free (not bound to platelets) serotonin (see Preparation of samples). NOTE: The direct determination of serotonin in PRP has shown that in about 10 % of the PRP samples unpredictable high serotonin concentrations were measured (results obtained by HPLC and fluorometry). To avoid such discrepancies, the seperate measurement of serotonin in platelets and PFP is recommended. Preparation of Reagents The contents of the kit can be divided into three separate runs. The volumes stated below are for one test procedure with 4 strips (32 determinations). If a larger number of strips is to be used the volumes have to be changed accordingly. 1. Assay Buffer 15 ml of the concentrate have to be diluted 1:10 with bidistilled water up to 150 ml. This gives the ready for use Assay Buffer. Store at 2-8 degrees Celsius. 2. Wash Buffer 15 ml of the concentrate have to be diluted 1:20 with bidistilled water up to 300 ml. This gives the ready for use Wash Buffer. Store at 2-8° Celsius. 3. Control Sera Add 0.5 ml of bidistilled water to each vial and let stand for 15 minutes. Mix by gentle swirling. For longer storage (> 1 day) freeze in aliquots. 4. Anti Biotin AP Dilute 60 µl of the concentrate with 6.0 ml of Assay Buffer (ready for use!). This gives the ready for use Anti Biotin AP. Prepare freshly prior to use! 5. PNPP Substrate Solution Dissolve 3 PNPP Substrate Tablets in 8 ml of PNPP Substrate Buffer. Dissolve the tablets approximately 10 minutes before the incubation with the Anti Biotin AP is finished. Sample Preparation After removing assay reagents from the refrigerator, allow them to reach room temperature before pipetting. Unused reagents should be stored at 2-8 degrees Celsius. Sample preparation should be performed in glass tubes. Standards and unknowns should be assayed in duplicate. Acylation of Samples and Controls The sample preparation (dilution and acylation) leads to a 207.25-fold (serum, urine, platelets, tissue homogenates, cell culture supernatants and a 23.5-fold (plasma) dilution of the samples, respectively. This has to be considered for the calculation of results. 1. Pipet 20 µl of Sample (serum, urine, platelet-extract, tissue homogenate) and Control Sera or 50 µl of platelet-free plasma (PFP) into glass test tubes. 2. Add 100 µl of Assay Buffer (ready for use!) to each tube and vortex mix. 3. Add 25 µl of Acylation Reagent 1 (3 %) (serum etc.) or 25 µl of Acylation Reagent 2 (10 %) (plasma) to each tube and vortex mix immediately. 4. Seal tubes and incubate for 15 minutes at 37° Celsius. 5. Add 4 ml (in case of serum etc.) or 1 ml (plasma) of Assay Buffer, vortex mix and remove the precipitated proteins by centrifugation (10 minutes at 1.500 x g). 6. Withdraw 50 µl aliquots for ELISA! Assay Procedure =============== 1. Pipette 50 µl of each Standard, acylated Control Serum and acylated patient samples into the appropriate wells. 2. Pipette 50 µl of Serotonin Biotin into the wells. 3. Pipette 50 µl of Antiserum into the wells and shake the plate carefully. 4. Seal the plate with an adhesive foil and incubate 16-20 h (over night) at 2-8 C. 5. Wash each well three times with Wash Buffer (the use of a washer is recommended!). Remove the Wash Buffer carefully. Invert plate and tap firmly on clean blotting paper to remove any remaining liquid. Note: The sensitivity and precision of this assay is markedly influenced by the correct performance of the washing procedure! 6. Pipette 150 µl of Anti Biotin AP into each well. 7. Seal with an adhesive foil and incubate for 120 minutes at room temperature on an orbital shaker (500 rpm). Prepare the Substrate Solution ten minutes before the incubation ends! 8. Wash each well three times with Wash Buffer (see above). 9. Pipette 200 µl PNPP Substrate Solution into each well. 10. Incubate at room temperature for 60 minutes on an orbital shaker (500 rpm). 11. Stop the substrate reaction by adding 50 µl of PNPP Stop Solution into all wells. Briefly mix contents by gently shaking the plate. 12. Read the optical density at 405 nm (reference wave length 600-650 nm) with a microtiterplate reader within 60 minutes after stopping.

Performance Characteristics

Back to Contents Assay Characteristics 1. Sensitivity The lowest detectable level that can be distinguished from the zero standard is 0.03 ng/ml (defined as 3 x standard deviation of the zero calibrator). 2. Specificity The cross reactivity of the anti-N-acylserotonin antiserum has been measured against various compounds. In all cases the interference from these compounds is unimportant due to the much higher levels of circulating serotonin. The percent cross reactivity is expressed as the ratio of serotonin concentration to the concentration of the reacting compound at 50 % binding of the zero standard. Compound Cross-reactivity(%) N-Acylated-Serotonin 100 Serotonin < 0.01 5-Hydroxytryptophol < 0.01 5-Hydroxy-3-indole acetic acid < 0.001 5-Hydroxytryptophan < 0.001 Melatonin < 0.1 3. Precision Intra-Assay-Variation (values in ng/ml) Mean Standard deviation % c.v n 111 12.1 10.9 32 413 26.3 6.4 32 Inter-Assay-Variation (values in ng/ml) Mean Standard deviation % c.v n 106 14.3 13.4 10 412 30.5 7.4 10 4. Recovery Normal human serum with a concentration of 0.09 g/ml was enriched with increasing amounts of serotonin. Added (g/ml) Expected (g/ml) Found (g/ml) Recovery (%) 2.000 2.090 2.040 97.6 1.000 1.090 1.008 92.4 0.500 0.590 0.554 93.8 0.250 0.340 0.318 93.5 0.125 0.215 0.198 92.0 0.066 0.156 0.146 93.5

Expected Values

Back to Contents Calculation of Results The optical density of the substrate blank is substracted from the optical densities of all standards and samples. On semilogarithmic graph paper the concentration of the standards (abscissa, logarithmic) are plotted against their corresponding optical density (ordinate, linear). Alternatively, the optical density of each standard and sample can be related to the optical density of the zero standard, expressed as the ratio OD/ODmax, and then plotted on the ordinate. The concentration of the samples can be read directly from this standard curve by using their average optical density. The sample preparation leads to a 207.25-fold (serum etc.) or 23.5-fold (plasma) dilution, respectively. Thus the values read from the standard curve have to be corrected as follows: Serum, urine, tissue homogenates, multiply by 207.25 cell culture supernatants Plasma (PFP) multiply by 23.5 This gives serotonin concentrations in ng/ml. Below is listed a typical example of a standard curve with the Serotonin- ELISA Conc. (ng/ml) OD1 OD2 mean value OD/ODmax (%) C.V. (%) 0.0 2.008 2.032 2.020 100.0 0.8 0.05 1.801 1.799 1.800 89.1 0.1 0.14 1.588 1.625 1.607 79.5 1.6 0.4 1.108 1.116 1.112 55.0 0.5 1.2 0.620 0.650 0.635 31.4 3.3 3.7 0.275 0.292 0.284 14.0 4.2 11 0.134 0.134 0.139 6.9 4.6 Calculation of serotonin in platelets 9 The content of serotonin in platelets is referred to 10 platelets. Following is given an example: Serotonin concentration : 50 ng/ml Number of platelets in PRP: 300.000 /µl equivalent to 30.000.000/100µl PRP and 200 µl of extraction volume. When using 20 µl for the test this is a platelet equivalent of 6 3 x 10 platelets. The serotonin content is referred to 1 ml. Therefore the used platelets equivalent of 20 µl has to be multiplied by 50. 6 9 3 x 10 x 50 = 0.15 x 10 platelets/ml with a serotonin content of 50 ng. The resulting serotonin content in the platelets is 9 9 9 333 ng/ 10 platelets (50 ng serotonin x 1.0 x 10 / 0.15 x 10 ). Expected values Serum It is recommended that each laboratory establishes its own range of normal serotonin values. A normal range study was performed with this assay principle. The following are the results: Serotonin reference values determined in human serum n 34 Mean (ng/ml) 92,6 SD 49,7 Range (ng/ml) up to 200 Platelet-free Plasma (PFP) n 35 Mean (ng/ml) 3,7 SD 1,5 Range (ng/ml) 1,8-7,5 Serotonin in platelets n 35 Mean (ng/10Mrd. platelets 490 SD 167 Range (ng/10Mrd. platelets) 217-861 Serotonin in 24 hours urine samples Serotonin: up to 200 µg/d Note: Certain foods contain substantial amounts of serotonin. Furthermore some medications may cause the release of serotonin and may lead to altered levels. Patients have to be abstained from such serotonin rich food (e.g. avocados, bananas, coffee, plums, pineapple, tomatoes, walnuts) as well as some medications (e.g. aspirin, corticotropin, MAO inhibitors, phenazetin, catecholamines, reserpin, nicotin).

Alternative Applications

Back to Contents

(More) Clinical Background

Back to Contents Ido P. Kema, Elisabeth G. E. de Vries, Albertine M. J. Schellings, Pieter E. Postmus, and Frits A. J. Muskiet Improved Diagnosis of Carcinoid Tumors by Measurement of Platelet Serotonin Clin. Chem. 38/4, 534-540 (1992) Carcinoid patients are diagnosed biochemically on the basis of increased urinary excretion of 5-hydroxyindoleacetic acid (5-HIAA); urinary and platelet serotonin concentrations are considered to provide complementary information. Using established HPLC methods with fluorometric detection, we evaluated the clinical usefulness of measurements of urinary 5-HIAA and urinary, plasma, and platelet serotonin in 30 consecutive patients with histologically proven carcinoid tumors of fore-, mid-, and hindgut origin before treatment. Ten patients showed no signs of serotonin overproduction; 14 had increased concentrations of urinary 5-HIAA and platelet serotonin; and platelet serotonin, but not urinary 5-HIAA, was increased in 6. None had increased urinary 5-HIAA excretion without an increase in platelet serotonin content. In cases with high rates of tumor serotonin secretion, platelet serotonin reached a maximum and did not correlate with serotonin secretion rate, whereas urinary 5-HIAA was correlated. Increased platelet serotonin was correlated with increased plasma serotonin and with occurence of carcinoid syndrome. Increased urinary serotonin, allegedly caused by increases in circulating 5-hydroxytryptophan, almost invariably coincided with increased platelet serotonin, but nor necessarily with above-normal urinary 5-HIAA excretion. From these results and long-term-monitoring of three patients during treatment, we conclude that platelet serotonin is more sensitive than urinary 5-HIAA for detecting carcinoids that secrete only small amounts of serotonin. --------------------------------------------------------------------------- Ido P. Kema, Elisabeth G. E. de Vries, Maarten J. H. Slooff, Bonne Biesma and Frits A. J. Muskiet Serotonin, Catecholamines, Histamine, and Their Metabolites in Urine, Platelets, and Tumor Tissue of Patients with Carcinoid Tumors Clin. Chem. 40/1, 86-95 (1994) We monitored long-term (median 11 months) concentrations of platelet serotonin and urinary serotonin, 5-hydroxyindoleacetic acid, and seven catecholamines metabolites in 44 patients with carcinoid tumors. Tumor serotonin and catecholamines contents (11 patients) and urinary histamine and N-methylhistamine (15 patients) were determined. Consistently increased concentrations of indoles, notably platelet serotonin, were observed in 96%, 43% and 0% of patients with mid-, fore-, and hindgut carcinoids, respectively. Urinary dopamine metabolites, notably 3-methoxytyramine, were consistently increased in 38%, 20%, and 7% of patients with mid-, hind-, anf foregut carcinoids, respectively. For urinary norepinephrine/epinephrine metabolites, notably normetanephrine and metanephrine, these data were 33%, 20%, and 14%, respectively. Midgut carcinoid tumors had the highest serotonin contents, whereas concentrations of catecholamines were independent of primary localization. There was no consistent relation between biogenic amine contents in tumors and urinary excretion of the amine metabolites. Occurence of carcinoid syndrome was related to increased serotonin production rate. Increased histamine production is not an important feature in patients with lung carcinoids or liver-metastasized ileum carcinoids. --------------------------------------------------------------------------- Elias V. Balakas, Gerasimos I. Bamihas, Michaelis Karamouzis, George Voyiatzis, Achilleas Tourkantonis. Histamine and Serotonin in Uremic Pruritus: Effect of Ondansetron in CAPD-Pruritic Patients Nephron 78, 395 - 402 (1998) Pruritus is a common, unpleasant symptom of uremic patients. Serotonin and histamine have been reported as possible mediators of uremic pruritus, and ondansetron is a potent and selective inhibitor of 5-HT3 receptors. The aims of our study were (1) to evaluate the effect of ondansetron on uremic pruritus in continuous ambulatory peritoneal dialysis (CAPD) patients and its safety and (2) to investigate the role of histamine and serotonin in uremic pruritus. To study the prevalence and pathogenesis of uremic pruritus, CAPD and hemodialysis (HD) patients were asked to complete a pruritus questionnaire. The replies were scored based on numerical scales, and the results were evaluated by the same investigator who did not know the patients. Pruritus was graded, according to the total points for each patient, as mild, moderate, or severe. Of 54 patients on HD, 29 (53.7 %) had pruritus, and of 43 patients on CAPD, pruritus was present in 21 (48.8 %). In HD patients, pruritus was mild in 14 (48.3 %), moderate in 12 (41.4 %), and severe in 3 (10.3 %) patients; the distribution in CAPD patients was 9 (42.9 %), 10 (47.6 %), and 2 (9.5 %), respectively. There was no correlation between the presence and severity of pruritus and age, sex, primary renal disease, duration of dialysis, dialysis solutions used, and hematological and biochemical parameters except for serum histamine and serotonin levels and their product. Plasma histamine levels in CAPD patients were 13.1 ñ 1.1 ng/ml in pruritic and 11.0 ñ 3.9 ng/ml in nonpruritic patients (p = 0.06), serum serotonin levels were 115.6 ñ 43.3 ng/ml and 64 ñ 42.3 ng/ml (p < 0.05), respectively, and the histamine x serotonin product was 1,461 ñ 576 and 646 ñ 545 (p < 0.01), respectively. Eleven CAPD patients (6 males, 5 females) with a mean age of 66 (range 33- 83) years and an average time on CAPD of 18 (range 3-31) months with moderate to severe pruritus were treated with ondansetron (4 mg twice daily p.o.) for a mean period of 3 (range 1-5) months. All patients responded to the treatment. There was a significant reduction of the severity of pruritus from the start of treatment, and on the 3rd day the pruritic score (mean value) was 10 (range 5-19) points, while at time 0 (before treatment) it was 26 (range 19-37) points (p < 0.0001). Pruritus disappeared in 7 patients at the end of the 1st week and in all patients at the end of the 2nd week of treatment. This effect was maintained during the study. Plasma histamine levels decreased significantly during the treatment from 12.9 ñ 1.2 to 6.7 ñ 5.9 ng/ml (p < 0.05). Also, serum serotonin levels were reduced from 125.1 ñ 47.8 to 59.3 ñ 27.5 ng/ml (p < 0.05) at the end of the 1st month of treatment, and the histamine x serotonin product showed a more significant reduction: from 1,544 ñ 656 to 454 ñ 436 (p < 0.01). Three patients reported an improvement in their nausea and vomiting during the treatment. Weekly clinical and laboratory examinations showed no side effects, adverse reactions, or other complications. Our data indicate that ondansetron is an effective, safe, and well- tolerated drug for the treatment of uremic pruritus in CAPD patients and that histamine and serotonin may have a crucial role in the appearance or perception of the uremic pruritus.

Sales Arguments

Back to Contents Potential customers and indications The clinical indications are mainly in the diagnosis of hypertension and diagnosis of carcinoid tumors as well as psychiatric disorders (see Guide-line to Biogenic Amines and IBL-information sheet). - Private laboratories, universities, research centres, psychiatric departments,pharmaceutical industry. Customers are medium-sized laboratories, hospitals with more than 800 beds, researchers in labs included in hospitals. So far, it makes no or only little sense to try to introduce the immunoassays in fully automated HPLC laboratories Arguments - Publication in: Journal of Musculoskeletal Pain, (1995), Abstracts, p. 65. Assay succesfully used in the diagnosis of fibromyalgia. Literature available on request. - Excellent correlation of the Serotonin ELISA to HPLC: y=-3.8 + 1.1x, r=0.99 - Close correlation between ELISA and RIA for serum samples:y=1.2x + 20, r=0.91 - Break apart strips for short assay runs - Assay applicable to serum, plasma, urine, tissue homogenates and cell culture supernatants as well as serotonin in platelets (detailed protocol) - Wide standard range; convenient measurement of pathological serum samples possible - Highly sensitive for the measurement of very low serotonin concentrations in different specimen e. g. in platelet free EDTA plasma - Assay has been succesfully used for the direct measurement of serotonin in platelets e. g. in scientific studies - Detailed normal values for serum, platelet-free plasma and serotonin in platelets. Competitors HPLC: very time-consuming, restricted to highly specialized laboratories General arguments HPLC vs. immunoassays: - After switching to immunoassays, the HPLC equipment may be used e. g. for therapeutic drug monitoring or for other analytes for which no routine test kits are available.

Product Literature

Back to Contents 1. Product Literature References 1.1. Balaskas, E., Bamihas, G., Karamouzis, M., Voyiatzis, G., Tourkantonis, A. Histamine and Serotonin in uremic pruritus: effect of ondansetron in CAPD-pruritic patients. Nephron, 78: 395-402 (1998) 1.2. Sprott, H., Kluge, H. Franke, S. Hein, G. (1995): Altered Serotonin-Levels in Patients with Fibromyalgia. In: Journal of Musculoskeletal Pain, Abstracts from the 3. World Congress on Myofascial Pain and Fibromyalgia, San Antonio, Texas, USA, Editor: I.J. Russel, p. 65. 1.3. Stratz, T., Samborski, W., Hrycai, P., Pap, T. Mackiewicz, S., Mennet, P., Mller, W. (1993): Die Serotoninkonzentration im Serum bei Patienten mit generalisierter Tendomyopathie (Fibromyalgie) und chronischer Polyarthritis. In: Medizinische Klinik, 88, 458-462. 2. Other Literature References Serotonin 1. Amir S et al. A role for serotonin in the circadian system revealed by the distribution of serotonin transporter and light-induced Fos immunoreactivity in the suprachiasmatic nucleus and intergeniculate leaflet. Neuroscience, 84: 1059-1073 (1998) . 2. D'Andrea G et al. Serotonin metabolism in cluster headache. Cephalalgia, 18: 94-96 (1998) . 3. Carrasco G et al. Plasma and platelet concentration and platelet uptake of serotonin in normal and pre-eclamptic pregnancies. Life Sci., 62: 1323-1332 (1998) . 4. Missouris CG et al. Serotonin and heart rate in hypertensive and normotensive subjects. Am. Heart J., 135. 838-843 (1998) . 5. Balaskas EV et al. Histamine and serotonin in uremic pruritus: effect of ondansetron in CAPD-pruritic patients. Nephron, 78: 395-402 (1998) 6. Huether G et al. Essen, Serotonin und Psyche. Dt. Žrztebl., 95: A-477-479 (1998) 7. Jovanovic S et al. Reference values of serotonin in urine and plasma determined by high-performance liquid chromatography with electrochemical detection. Clin. Lab., 44: 263-268 (1998) 8. Karamouzis M et al. Histamine and serotonin as markers of uremic pruritus. Clin. Chem., 43: S113 (1997) . 9. Joshi G et al. Platelet serotonin kinetics in depressed adolescents. Biogenic Amines, 13: 591-602 (1997) 10.Chandra M et al. Platelets in pathogenesis of primary hypertension. Biogenic Amines, 13: 547-563 (1997) 11.Pussard E et al. Validation of HPLC-amperometric detection to measure serotonin in plasma, platelets, whole blood, and urine. Clin. Chem., 42: 1086-1091 (1996) 12.Lechin F et al. Increased levels of free serotonin in plasma of symptomatic asthmatic patients. Ann. Allergy Asthma Immunol., 77: 245-253 (1996) . 13.Sprott H et al. Altered serotonin-levels in patients with fibromyalgia. J. Musculoskeletal Pain, 3: 65 (1995) . 14.Owens MJ and Nemeroff CB. Role of serotonin in the pathophysiologie of depression: focus on the serotonin transporter. Clin. Chem., 40: 288-295 (1994) 15.Kema IP et al. Serotonin, catecholamines, histamine, and their metabolites in urine, platelets, and tumor tissue of patients with carcinoid tumors. Clin. Chem., 40: 86-95 (1994) 16.Stratz T et al. Die Serotoninkonzentration im Serum bei Patienten mit generalisierter Tendomyopathie (Fibromyalgie) und chronischer Polyarthritis. Medizinische Klinik, 88: 458-462 (1993) 17.Middelkoop CM et al. Platelet-poor plasma serotonin in normal and preeclamptic pregnancy. Clin. Chem., 39. 1675-1678 (1993) 18.Hindberg I and Naesh O. Serotonin concentrations in plasma and variations during the menstrual cycle. Clin. Chem., 38: 2087-2089 (1993) 19.Kema IP et al. Influence of a serotonin- and dopamine-rich diet on platelet serotonin content and urinary excretion of biogenic amines and their metabolites. Clin. Chem., 38: 1730-1736 (1992) 20.Kema IP et al. Improved diagnosis of carcinoid tumors by measurement of platelet serotonin. Clin. Chem., 38: 534-540 (1992) 21.Quintana J. Platelet serotonin and plasma tryptophan decreases in endogenous depression. Clinical, therapeutic, and biological correlations. J. Affective Disorders, 24: 55-62 (1992) 22.Chauveau J et al. Rapid specific enzyme immunoassay of serotonin. Clin Chem., 37: 1178-1184 (1991) 23.Martin F et al. Plasma serotonin: factors affecting its levels. Biogenic Amines, 7: 57-62 (1990) 24.Flachaire E et al. Determination of reference values for serotonin concentration in platelets of healthy newborns, children, adults, and elderly subjects by HPLC with electrochemical detection. Clin. Chem., 36: 2117-2120 (1990) 25.Miya TG and Wu JT. Development of whole blood and serum assay for serotonin by high-performance liquid chromatography (HPLC). Clin Chem., 36: 1153 (1990) . 26.Anthony M and Lance JW. Plasma serotonin in patients with chronic tension headaches. J. Neurol. Neurosurg. Psychiatry, 52: 182-184 (1989) 27.Marasini B et al. Platelet and plasma serotonin in patients with liver cirrhosis. J. Clin. Chem. Clin. Biochem., 27: 419-421 (1989) 28.Vollrath L et al. Serotonin and melatonin contents in the pineal glands from different stocks and strains of laboratory rats. Z. Versuchstierkd., 32: 57-63 (1989) 29.Singh SM et al. Concentrations of serotonin in plasma - a test for appendicitis? Clin Chem., 34: 2572-2574 (1988) 30.Koyama E et al. Simultaneous determination of four monoamine metabolites and serotonin in cerebrospinal fluid by "high-performance" liquid chromatography with electrochemical detection; application for patients with Alzheimer's disease. Clin Chem., 34: 680-684 (1988) 31.Imai Y et al. Simultaneous determination of catecholamines and serotonin by liquid chromatography, after treatment with boric acid gel. Clin. Chem., 34: 528-530 (1988) 32.Richter G et al. Serotonin release into blood after food and pentagastrin. Gastroenterol., 91: 612-618 (1986) 33.Mailman RB and Kilts CD. Analytical considerations for quantitative determination of serotonin and its metabolically related products in biological matrices. Clin. Chem., 31: 1849-1854 (1985) 34.Ingebretsen OC et al. Liquid chromatography of serotonin and adenine nucleotides in blood platelets, illustrated by evaluation of functional integrity of platelet preparations. Clin. Chem., 31: 695-698 (1985) 35.Picard M et al. Determination of serotonin in plasma by liquid chromatography with electrochemical detection. J. Chromatography, 341: 445-451 (1985)
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