Plasma Metabolites In Relation Research Articles

Strategies for Using Postcolumn Infusion of Standards to Correct for Matrix Effect in LC-MS-Based Quantitative Metabolomics.

The matrix effect limits the accuracy of quantitation of the otherwise popular metabolomics technique liquid chromatography coupled to mass spectrometry (LC-MS). The gold standard to correct for this phenomenon, whereby compounds coeluting with the analyte of interest cause ionization enhancement or suppression, is to quantify an analyte based on the peak area ratio with an isotopologue added to the sample as an internal standard. However, these stable isotopes are expensive and sometimes unavailable. Here, we describe an alternative approach: matrix effect correction and quantifying analytes using a signal ratio with a postcolumn infused standard (PCIS). Using an LC-MS/MS method for eight endocannabinoids and related metabolites in plasma, we provide strategies to select, optimize, and evaluate PCIS candidates. Based on seven characteristics, the structural endocannabinoid analogue arachidonoyl-2'-fluoroethylamide was selected as a PCIS. Three methods to evaluate the PCIS correction vs no correction showed that PCIS correction improved values for the matrix effect, precision, and dilutional linearity of at least six of the analytes to within acceptable ranges. PCIS correction also resulted in parallelization of calibration curves in plasma and neat solution, for six of eight analytes even with higher accuracy than peak area ratio correction with their stable isotope labeled internal standard, i.e., the gold standard. This enables quantification based on neat solutions, which is a significant step toward absolute quantification. We conclude that PCIS has great, but so far underappreciated, potential in accurate LC-MS quantification.

Causal relationships between immunophenotypes, plasma metabolites, and temporomandibular disorders based on Mendelian randomization

While numerous studies have underscored the implication of immune cells and metabolites in temporomandibular disorders (TMD), conclusive evidence for causality remains elusive. Consequently, our aim is to explore the causal connections between immunophenotypes and plasma metabolites in relation to TMD employing a bidirectional Mendelian randomization (MR) approach. Summary statistics data on 731 immunophenotypes (n = 3757) and 1091 plasma metabolites (n = 8299) were obtained from comprehensive genome-wide association studies (GWAS), while TMD data (5668 cases and 205,355 controls) were acquired from the FinnGen Consortium. Bidirectional MR analyses and a two-step MR approach assessed causal relationships and potential intermediaries. Various corrections and sensitivity analyses were utilized to assess the robustness of the findings. Two immunophenotypes and seven metabolites were significantly associated with TMD risk. Specifically, Alpha-hydroxyisovalerate mediated the link between CD33 on CD33dim HLA DR + CD11b + and TMD (β = 0.034, P = 5.95 × 10–5), while CD8 on NKT cells mediated the causal relationship between 5-acetylamino-6-formylamino-3-methyluracil levels and TMD (β = 0.069, P = 5.11 × 10–5). Our findings revealed the causal relationships between immunophenotypes and plasma metabolites on TMD from a genetic perspective, potentially aiding in TMD prevention.

Association between maternal plasma one-carbon biomarkers during pregnancy and fetal growth in twin pregnancies

Objective: To characterize the relationship between the levels of plasma methyl donor and related metabolites (including choline, betaine, methionine, dimethylglycine and homocysteine) and fetal growth in twin pregnancies. Methods: A hospital-based cohort study was used to collect clinical data of 92 pregnant women with twin pregnancies and their fetuses who were admitted to Peking University Third Hospital from March 2017 to January 2018. Fasting blood was collected from the pregnant women with twin pregnancies (median gestational age: 18.9 weeks). The levels of methyl donors and related metabolites in plasma were quantitatively analyzed by high-performance liquid chromatography combined with mass spectrometry. The generalized estimation equation was used to analyze the relationship between maternal plasma methyl donors and related metabolites levels and neonatal outcomes of twins, and the generalized additive mixed model was used to analyze the relationship between maternal plasma methyl donors and related metabolites levels and fetal growth ultrasound indicators. Results: (1) General clinical data: of the 92 women with twin pregnancies, 66 cases (72%) were dichorionic diamniotic (DCDA) twin pregnancies, and 26 cases (28%) were monochorionic diamniotic (MCDA) twin pregnancies. The comparison of the levels of five plasma methyl donors and related metabolites in twin pregnancies with different basic characteristics showed that the median levels of plasma choline and betaine in pregnant women ≥35 years old were higher than those in pregnant women <35 years old, and the differences were statistically significant (all P<0.05). (2) Correlation between plasma methyl donor and related metabolites levels and neonatal growth indicators: after adjusting for confounding factors, plasma homocysteine level in pregnant women with twins was significantly negatively correlated with neonatal birth weight (β=-47.9, 95%CI:-94.3- -1.6; P=0.043). Elevated methionine level was significantly associated with decreased risks of small for gestational age infants (SGA; OR=0.5, 95%CI: 0.3-0.9; P=0.021) and low birth weight infants (OR=0.6, 95%CI: 0.4-0.9; P=0.020). Increased homocysteine level was associated with increased risks of SGA (OR=1.5, 95%CI: 1.0-2.2; P=0.029) and inconsistent growth in twin fetuses (OR=1.9, 95%CI: 1.0-3.7; P=0.049). (3) Correlation between the levels of plasma methyl donors and related metabolites and intrauterine growth indicators of twins pregnancies: for every 1 standard deviation increase in plasma choline level in pregnant women with twin pregnancies, fetal head circumference, abdominal circumference, femoral length and estimated fetal weight in the second trimester increased by 1.9 mm, 2.6 mm, 0.5 mm and 20.1 g, respectively, and biparietal diameter, abdominal circumference and estimated fetal weight increased by 0.7 mm, 3.0 mm and 38.4 g in the third trimester, respectively, and the differences were statistically significant (all P<0.05). (4) Relationship between plasma methyl donor and related metabolites levels in pregnant women with different chorionicity and neonatal birth weight and length: the negative correlation between plasma homocysteine level and neonatal birth weight was mainly found in DCDA twin pregnancy (β=-65.9, 95%CI:-110.6- -21.1; P=0.004). The levels of choline, betaine and dimethylglycine in plasma of MCDA twin pregnancy were significantly correlated with the birth weight and length of newborns (all P<0.05). Conclusion: Homocysteine level is associated with low birth weight in twins, methionine is associated with decreased risk of SGA, and choline is associated with fetal growth in the second and third trimesters of pregnancy.

Integrated Multi-Omics Analysis for Inferring Molecular Players in Inclusion Body Myositis.

Inclusion body myositis (IBM) is an acquired inflammatory myopathy affecting proximal and distal muscles that leads to weakness in patients over 50. It is diagnosed based on clinical and histological findings in muscle related to inflammation, degeneration, and mitochondria. In relation to IBM, a shortage of validated disease models and a lack of biomarkers and effective treatments constitute an unmet medical need. To overcome these hurdles, we performed an omics analysis of multiple samples from IBM patients (saliva, fibroblasts, urine, plasma, and muscle) to gain insight into the pathophysiology of IBM. Degeneration was evident due to the presence of amyloid β peptide 1-42 (Aβ1-42) in the saliva of the analyzed IBM patients. The presence of metabolic disarrangements in IBM was indicated by an imbalanced organic acid profile in fibroblasts and urine. Specifically, abnormal levels of L-pyroglutamic and orotic acid were supported by the abnormal expression of related metabolites in plasma and urine (glutathione and pyrimidines) and the aberrant expression of upstream gene regulators (L2HGDH, IDH2, OPLAH, and ASL) in muscle. Combined levels of L-pyroglutamic and orotic acid displayed an outstanding biomarker signature in urine with 100% sensitivity and specificity. The confirmation of systemic metabolic disarrangements in IBM and the identification of novel biomarkers reported herein unveil novel insights that require validation in larger cohorts.

Distinct composition and metabolic functions of human gut microbiota are associated with cachexia in lung cancer patients

Cachexia is associated with decreased survival in cancer patients and has a prevalence of up to 80%. The etiology of cachexia is poorly understood, and limited treatment options exist. Here, we investigated the role of the human gut microbiome in cachexia by integrating shotgun metagenomics and plasma metabolomics of 31 lung cancer patients. The cachexia group showed significant differences in the gut microbial composition, functional pathways of the metagenome, and the related plasma metabolites compared to non-cachectic patients. Branched-chain amino acids (BCAAs), methylhistamine, and vitamins were significantly depleted in the plasma of cachexia patients, which was also reflected in the depletion of relevant gut microbiota functional pathways. The enrichment of BCAAs and 3-oxocholic acid in non-cachectic patients were positively correlated with gut microbial species Prevotella copri and Lactobacillus gasseri, respectively. Furthermore, the gut microbiota capacity for lipopolysaccharides biosynthesis was significantly enriched in cachectic patients. The involvement of the gut microbiome in cachexia was further observed in a high-performance machine learning model using solely gut microbial features. Our study demonstrates the links between cachectic host metabolism and specific gut microbial species and functions in a clinical setting, suggesting that the gut microbiota could have an influence on cachexia with possible therapeutic applications.

A diet-induced gut microbiota component and related plasma metabolites are associated with depressive-like behaviour in rats

It is well-established in preclinical studies that various probiotics may improve behaviours related to psychiatric disease. We have previously shown that probiotics protected against high-fat diet (HFD)-induced depressive-like behaviour in Flinders Sensitive Line (FSL) rats, whereas FSL rats on control (CON) diet were unaffected. Therefore, we hypothesised that a dysmetabolic component of depression may exist that involves the gut microbiota and that such component may be reflected in the plasma metabolome.The aims of the present study post hoc analyses were 1) to study the effect of probiotics on gut microbiota composition and its association with depressive-like behaviour in FSL rats, and 2) to identify plasma metabolites associated with gut microbiota and depressive-like behaviour.Forty-six FSL rats were fed CON or HFD and treated with multi-species probiotics (nine Bifidobacterium, Lactococcus and Lactobacillus species) for 12 weeks. Faecal samples were collected for 16S rRNA (VR4) gene amplicon sequencing (Illumina MiSeq), and an untargeted plasma metabolomics was performed.We found that probiotics increased the relative faecal abundance of the Bifidobacterium, Lactococcus and Lactobacillus genera in HFD-fed rats only. Also, a HFD-induced microbiota component associated with depressive-like behaviour was identified, and probiotics improved the component score. Finally, the plasma levels of 44 metabolites correlated with the depression-related microbiota component, and three such metabolites had good predictive ability for depressive-like behaviour.Potentially, our findings imply that a subtype of depression characterised by a diet-induced, pro-depressant gut microbiota may exist and that analysis of related plasma metabolites may reveal aberrant microbiota functioning related to depression.

Habitual Sleep and human plasma metabolomics.

Sleep plays an important role in cardiometabolic health. The sleep-wake cycle is partially driven by the endogenous circadian clock, which governs a range of metabolic pathways. The association between sleep and cardiometabolic health may be mediated by alterations of the human metabolome. To better understand the biological mechanism underlying the association between sleep and health, we examined human plasma metabolites in relation to sleep duration and sleep timing. Using an untargeted approach, 329 fasting plasma metabolites were measured in 277 Chinese participants. We measured sleep timing (midpoint between bedtime and wake up time) using repeated time-use surveys (4 weeks during one year) and previous night sleep duration from questionnaires completed before sample donation. We found 64 metabolites that were associated with sleep timing with a false discovery rate of 0.2 or lower, after adjusting for potential confounders. Notably, we found that later sleep timing was associated with higher levels of multiple metabolites in amino acid metabolism, including branched chain amino acids and their gamma-glutamyl dipeptides. We also found widespread associations between sleep timing and numerous metabolites in lipid metabolism, including bile acids, carnitines and fatty acids. In contrast, previous night sleep duration was not associated with plasma metabolites in our study. Sleep timing was associated with a large number of metabolites across a variety of biochemical pathways. Some metabolite associations are consistent with a relationship between late chronotype and adverse effects on cardiometabolic health.

Relationship between salivary/pancreatic amylase and body mass index: a systems biology approach

BackgroundSalivary (AMY1) and pancreatic (AMY2) amylases hydrolyze starch. Copy number of AMY1A (encoding AMY1) was reported to be higher in populations with a high-starch diet and reduced in obese people. These results based on quantitative PCR have been challenged recently. We aimed to re-assess the relationship between amylase and adiposity using a systems biology approach.MethodsWe assessed the association between plasma enzymatic activity of AMY1 or AMY2, and several metabolic traits in almost 4000 French individuals from D.E.S.I.R. longitudinal study. The effect of the number of copies of AMY1A (encoding AMY1) or AMY2A (encoding AMY2) measured through droplet digital PCR was then analyzed on the same parameters in the same study. A Mendelian randomization analysis was also performed. We subsequently assessed the association between AMY1A copy number and obesity risk in two case-control studies (5000 samples in total). Finally, we assessed the association between body mass index (BMI)-related plasma metabolites and AMY1 or AMY2 activity.ResultsWe evidenced strong associations between AMY1 or AMY2 activity and lower BMI. However, we found a modest contribution of AMY1A copy number to lower BMI. Mendelian randomization identified a causal negative effect of BMI on AMY1 and AMY2 activities. Yet, we also found a significant negative contribution of AMY1 activity at baseline to the change in BMI during the 9-year follow-up, and a significant contribution of AMY1A copy number to lower obesity risk in children, suggesting a bidirectional relationship between AMY1 activity and adiposity. Metabonomics identified a BMI-independent association between AMY1 activity and lactate, a product of complex carbohydrate fermentation.ConclusionsThese findings provide new insights into the involvement of amylase in adiposity and starch metabolism.

Identifying novel plasma biomarkers of pancreatic beta‐cell function using acute and chronic models of hyperglycemia.

Beta‐cell function is impaired in type 2 diabetes. Identifying related plasma metabolites related to insulin secretion may help identify novel treatment targets. Intravenous glucose‐stimulated insulin secretion (GSIS) was measured in healthy subjects before and after acute 24‐hour experimental‐elevation of plasma glucose (+5 mM). GSIS was also measured in age/BMI‐matched type 2 diabetic (T2D) patients. The plasma metabolite profile (358 compounds) was determined using LC/GC‐MS and correlated with group differences in GSIS and with hyperglycemia‐induced changes in GSIS. GSIS was lower in T2D compared to controls (6.8±0.8 vs. 10.1±0.9 pmol/kg/min; P&lt;0.05), but elevated following experimental hyperglycemia in healthy controls (13.1±0.8). Hexadecanedioate, octadecanedioate and 3‐aminoisobutyrate were significantly correlated with GSIS (r= −0.66,−0.61,−0.56; P&lt;0.05) showing the same direction of change in both acute and chronic models of hyperglycemia. These plasma metabolites are biomarkers of beta‐cell function in humans and may reveal novel therapeutic targets. The effect of these compounds on GSIS in perifused pancreatic islets is currently being determined.

Generation of an index for physiological imbalance and its use as a predictor of primary disease in dairy cows during early lactation

Physiological imbalance (PI) is a situation in which physiological parameters deviate from the normal and cows consequently have an increased risk of developing production diseases and reduced production or reproduction. The objectives of this work were (1) to generate an index for PI based on several plasma metabolites and (2) to compare the use of this index with calculated energy balance (EBAL) and individual plasma metabolites in relation to risk of disease during early lactation. We used a total of 634 lactations from 317 cows consisting of 3 breeds ranging from a parity of 1 to 4. Weekly blood samples were analyzed for selected metabolites; that is, urea nitrogen, albumin, cholesterol, nonesterified fatty acids (NEFA), glucose, and β-hydroxybutyrate (BHBA). Energy intake and EBAL were calculated; veterinary treatment records and daily composite milk somatic cell counts were used to determine incidence of disease. Data were adjusted for numerous fixed effects (e.g., parity, breed, and week around calving) before further statistical analysis. The time of disease (TOD) was recorded as the day in which the signs of disease were observed (TOD=0). The week before and after TOD was ± n wk relative to TOD=0. Each week, all plasma metabolites were individually adjusted to an overall mean (=0) and variance (=1). The normalized variables were included in regression analyses by week of lactation to identify metabolites that explain the variation in calculated EBAL, as a reflection of degree of PI. Nonesterified fatty acids, BHBA, and glucose were weighted within each week based on regression coefficients (i.e., x1–x3 below) generated from a model to predict EBAL. Data from wk −1 relative to TOD were analyzed using a mixed linear model to relate degree of PI and metabolites in blood to risk of disease. The weekly PI index was defined as PI=(x1 × [NEFA])+x2 × [BHBA] − x3 × [glucose])/3. For diseases that developed ≥2 wk after calving, no variables were associated with risk of disease. Prepartal PI and plasma NEFA were better predictors of disease (i.e., metritis, retained placenta, and milk fever) at wk 1 than EBAL and plasma BHBA and glucose. Examining the relationship between PI and milk constituents is needed for the development of an automated in-line and real-time surveillance system for early detection of risk animals on-farm.

HPLC analysis of asymmetric dimethylarginine (ADMA) and related arginine metabolites in human plasma using a novel non-endogenous internal standard

Background Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthesis which has been implicated in the endothelial dysfunction. Methods for ADMA measurement often yield widely differing results, and few methods simultaneously offer satisfactory accuracy and precision. We describe a fully validated HPLC method for analysis of arginine and its methylated derivatives in human plasma using a novel internal standard. Methods Arginine and related metabolites are extracted from plasma by solid phase extraction (SPE), derivatised with ortho-phthaldialdehyde and separated by isocratic reverse phase chromatography. Monoethylarginine (MEA), which is not endogenously present in human plasma was used as internal standard. SPE and chromatographic procedures are optimised and recovery, precision, linearity and sensitivity of the assay established. The suitability and performance of MEA is compared with that of monomethylarginine (MMA), the internal standard most commonly used in HPLC methods. Results SPE yields high and reproducible recoveries (> 90%). The analytes of interest are chromatographically well resolved. The method has high sensitivity (LOD, 0.01 μmol/L for arginine and 0.001 μmol/L for ADMA, SDMA and homoarginine) and good precision (CV, 2.5% for ADMA). The data obtained with the internal standards MEA and MMA is comparable in terms of assay precision and population reference intervals. Conclusions We describe an optimised isocratic HPLC method for the simultaneous measurement of arginine and related metabolites in plasma which exhibits satisfactory precision and is suitable for routine use. Its main advantage over other published HPLC methods is the use of the novel internal standard, MEA, which unlike other commonly used internal standards is not inherent in human plasma.

Orally administered rosmarinic acid is present as the conjugated and/or methylated forms in plasma, and is degraded and metabolized to conjugated forms of caffeic acid, ferulic acid and m-coumaric acid

Rosmarinic acid (RA) is contained in various Lamiaceae herbs used commonly as culinary herbs. Although RA has various potent physiological actions, little is known on its bioavailability. We therefore investigated the absorption and metabolism of orally administered RA in rats. After being deprived of food for 12 h, RA (50 mg/kg body weight) or deionized water was administered orally to rats. Blood samples were collected from a cannula inserted in the femoral artery before and at designated time intervals after administration of RA. Urine excreted within 0 to 8 h and 8 to 18 h post-administration was also collected. RA and its related metabolites in plasma and urine were measured by LC-MS after treatment with sulfatase and/or β-glucuronidase. RA, mono-methylated RA (methyl-RA) and m-coumaric acid (COA) were detected in plasma, with peak concentrations being reached at 0.5, 1 and 8 h after RA administration, respectively. RA, methyl-RA, caffeic acid (CAA), ferulic acid (FA) and COA were detected in urine after RA administration. These components in plasma and urine were present predominantly as conjugated forms such as glucuronide or sulfate. The percentage of the original oral dose of RA excreted in the urine within 18 h of administration as free and conjugated forms was 0.44 ± 0.21% for RA, 1.60 ± 0.74% for methyl-RA, 1.06 ± 0.35% for CAA, 1.70 ± 0.45% for FA and 0.67 ± 0.29% for COA. Approximately 83% of the total amount of these metabolites was excreted in the period 8 to 18 h after RA administration. These results suggest that RA was absorbed and metabolized as conjugated and/or methylated forms, and that the majority of RA absorbed was degraded into conjugated and/or methylated forms of CAA, FA and COA before being excreted gradually in the urine.

Effects of incremental changes in forage: concentrate ratio on plasma hormone and metabolite concentrations and products of rumen fermentation in fattening beef steers

AbstractAs part of an investigation of factors responsible for a previously reported lower efficiency of carcass lean gain in steers offered grass silage diets, 16 Simmental × Friesian steers (515 (s.e. 6·4) kg) were offered perennial ryegrass silage ad libitum (C0) or silage plus rolled barley at 200 (C20), 400 (C40) or 600 (C60) g/kg total diet dry matter (DM). Barley-supplemented diets were intake-restricted to provide equal DM and metabolizable energy (ME) intakes to those offered C0. Eight steers were selected at random to determine the ME contents of the diets by open-circuit respiration calorimetry. The other eight steers were offered the same diets and were blood-sampled at 20- to 60-min intervals, for 10 h, to monitor changes in the concentrations of a number of nutritionally related plasma metabolites and hormones. Estimated ME intakes in these steers were 85·7, 83·1, 84·4 and 86·2 (s.e. 0·91) MJ/day from diets C0, C20, C40 and C60 respectively. Rumen-fistulated Hereford × Friesian steers provided 24-h rumen data for the same diets offered at equal amounts of ME per kg metabolic live weight.Mean 24-h plasma concentrations of insulin and insulin-like growth factor 1 (IGF-1) were linearly and positively related (P &lt; 0·01 and P &lt; 0·001 respectively) and glucagon quadratically related (P &lt; 0·05) to the proportion of barley in the diet. Plasma insulin increased after feeding on all diets but concentrations on diets C40 and C60 were significantly higher than those on C0 and C20 at all post-feeding sampling times up to 9 h after feeding. Plasma IGF-1 concentrations increased above pre-feeding levels following feeding of the higher barley diets (C40 and C60; P = 0·053) but remained unchanged in steers offered C0 and C20. Mean plasma concentrations of glucose were unaffected by diet but those of β-hydroxybutyrate (BOHB) and urea were positively and negatively related respectively (both P &lt; 0·001) to the proportion of barley in the diet. Plasma BOHB and urea concentrations also changed with time after feeding (P &lt; 0·001). Amongst the rumen parameters measured (pH; ammonia and volatile fatty acid concentrations and proportions) only the mean 24-h concentrations and proportions of butyrate were positively related to the proportion of barley in the diet (P = 0·051 and P &lt; 0·05 respectively). All rumen parameters were affected by time after feeding (acetate, P &lt; 0·01; others, P &lt; 0·001) but there was no interaction between treatment and time for any parameter.

Concentration of ceftiofur metabolites in the plasma and lungs of horses following intramuscular treatment

Ceftiofur sodium, a broad spectrum cephalosporin antibiotic approved for veterinary use, is metabolized to desfuroylceftiofur which is conjugated to micro as well as macromolecules. Twelve horses, weighting 442-618 kg, were injected intramuscularly with a single dose of 2.2 mg ceftiofur/kg (1.0 mg/lb) body weight. Blood was collected at various intervals over 24 h after treatment. Three groups of four horses each were euthanized and lungs were collected at 1, 12, and 24 h after treatment. The concentration of desfuroylceftiofur and desfuroylceftiofur conjugates in the plasma and lungs was determined by converting them to desfuroylceftiofur acetamide (DCA) and measured DCA by high performance liquid chromatography with UV detection. The average maximum concentration (Cmax) of desfuroylceftiofur and related metabolites in plasma expressed as ceftiofur equivalents was 4.46 +/- 0.93 micrograms/ml occurred at 1.25 +/- 0.46 h after treatment. These concentrations declined to 0.99 +/- 0.16, 0.47 +/- 0.15 and 0.17 +/- 0.02 microgram/ml at 8, 12, and 24 h, respectively. The mean residence time of ceftiofur metabolites was 6.10 +/- 1.27 h. Concentrations of desfuroylceftiofur and desfuroylceftiofur conjugates in the lungs of horses expressed as ceftiofur equivalents were 1.40 +/- 0.36, 0.27 +/- 0.07, and 0.15 +/- 0.08 micrograms/ml at 1, 12, and 24 h, respectively. These concentrations of the drug at 12 and 24 h in lung homogenate were similar but slightly lower than plasma concentrations in the same horses, and the plasma pharmacokinetic values including half-life were similar to those observed at the approved dose of 1.1-2.2 mg ceftiofur/kg body weight administered intramuscularly once daily for 3-5 days in cattle.

Relationship Between Serotoninergic Measures in Blood and Cerebrospinal Fluid Simultaneously Obtained in Humans

The relationships between the concentration of serotonin (5-HT) and related metabolites in human blood and CSF have been studied. Plasma tryptophan (TP), 5-HT, 5-hydroxyindoleacetic acid (5-HIAA), and indoleacetic acid (IAA), whole-blood 5-HT, and CSF TP, 5-HT, 5-HIAA, IAA, homovanillic acid, and 3-methoxy-4-hydroxyphenylethylene glycol were determined in 35 unmedicated outpatients who underwent minor surgical operations and had no history of psychiatric or neurological illnesses. Significant correlations were found between the serotoninergic parameters analyzed in blood and CSF. Plasma free 5-HT correlated significantly with CSF 5-HT (r = 0.411, p less than 0.02), and plasma 5-HIAA correlated with the CSF 5-HIAA/5-HT ratio (r = 0.508, p less than 0.004). The concentration of 5-HIAA in CSF correlated with the plasma 5-HIAA/5-HT ratio (r = 0.405, p less than 0.026) (which can be taken as an index of monoamine oxidase type A activity in peripheral tissues) and with the platelet 5-HT/plasma 5-HT ratio (r = 0.375, p less than 0.05). The concentrations of IAA in CSF and plasma were strongly correlated (r = 0.899, p less than 0.001). The significance of these results and their relationship to the use of "in vivo" measures of 5-HT and related metabolites in plasma and platelets as an index of serotoninergic function in affective disorders are discussed.