Small Volumes Of Urine Research Articles

CYSTITIS IN MIX BREED DOG

Cystitis is an inflammation of the vesica urinaria. A male mixed breed dog named Max, 3 years old and weighing 18.68 kg, has stranguria and dysuria with a small volume of urine, and hematuria. The purpose of this report is to discuss the incidence of cystitis in male mixed breed dogs in performing diagnostic measures as well as appropriate treatment and therapy for the dog. Clinical examination showed pulse frequency, heart rate, respiratory, and body temperature within the normal range. Abdominal palpation revealed signs of pain. Routine hematology analysis revealed struvite and tyrosine crystals. Urinalysis showed elevated leucocytes and post-renal proteinuria. Urine culture examination revealed Staphylococcus sp. Ultrasonography examination showed thickening of the vesical urinary mucous. Based on findings, the dog case was diagnosed with cystitis. Treatment included antibiotics amoxicillin, anti-inflammatory drug dexamethasone, and additional herbal medicine kejibeling. The case dog showed improvement after seven days of treatment with no symptoms such as haematuria. To minimise the recurrence of cystitis in dogs, Urinary Care feed and ad libitum watering can be done to help dissolve crystals in the vesica urinaria.

STRUVITE CRYSTAL UROLITHIASIS IN A 2 YEAR OLD CASTRATED MALE CAT

Urolithiasis, kidney failure, urinary tract infections are examples of disorders of the urinary tract that often become problems in cats. Urolithiasis is a medical term to describe the presence of stones or crystals known as “urolith” in the urinary tract. Inappropriate composition and feeding methods can cause nutritional imbalances in the cat's body, which can affect the acidity level (pH) of urine, urine volume and urine concentration which can cause the formation of excessive minerals in the urine. The aim of writing this article is to provide information regarding the treatment of crystal urolithiasis in local cats. A male cat named Goblin, has been castrated, has colored hair orange with body weight 5.3 kg, experiencing difficulty urinating since 3 weeks, excreting a small volume of urine, and blood in the urine. On physical examination the cat always curled up and felt pain when palpation was carried out abdominal part. The results of the cat's blood test showed leukocytosis, lymphocytosis, thrombocytopenia, and RDW CV increased and PCT decreased. The results of the cat's urinalysis showed presence of leukocytes +2, Protein +1, pH 8.0, specific gravity 1.030, Calcium +1, Microalbumin +1.Sediment test results also found the presence of deposits of magnesium ammonium phosphate crystal particles (struvite crystals).Results of urine culture examination bacteria were foundStaphylococcus sp. Ultrasound results showed enlargement of the left kidney and deposits of hyperechoic particles (crystals) accompanied by enlargement of the uterine sac. The cat was diagnosed with urolithiasis with a fausta prognosis. The cat was treated by installing a 4Fr/1.3x130mm urinary catheter and flushing using 0.9% physiological NaCl to make it easier for the cat to urinate. The therapy given is the antibiotic ciprofloxacin hcl 5 mg/kg BW s500 mg preparation with a dose of 50 mg/kg BW per oral (PO) administered 2 times a day for seven days, administering the anti-inflammatory dexamethasone 0.5 mg with a dose of 1 mg/kg BW PO and given 2 times a day for four days, and kejibeling herbal medicine 1 tablet times a day for seven days. Cats are also given feed therapy in the form of Royal Canin Urinary S/O for 3 months. Evaluation of the cat's condition improved after therapy for 7 days. There is a need to educate clients regarding cat care, feeding and drinking water. Therefore, the cat's feed in cases must be replaced by providing urinary therapy feed.

Monitoring Exposure to Five Chemical Warfare Agents Using the Dried Urine Spot Technique and Liquid Chromatography-Mass Spectrometry/Mass Spectrometry—In Vivo Determination of Sarin Metabolite in Mice

Dried urine spot (DUS) is a micro-sample collection technique, known for its advantages in handling, storage and shipping. It also uses only a small volume of urine, an essential consideration in working with small animals, or in acute medical situations. Alkyl-phosphonic acids are the direct and indicative metabolites of organophosphorus chemical warfare agents (OP-CWAs) and are present in blood and urine shortly after exposure. They are therefore crucially important for monitoring casualties in war and terror scenarios. We report here a new approach for the determination of the metabolites of five CWAs in urine using DUS. The method is based on a simple and rapid sample preparation, using only 50 µL of urine, spotted and dried on DBS paper, extracted using 300 µL methanol/water and analyzed via targeted LC-MS/MS. The detection limits for the five CWAs, sarin (GB), soman (GD), cyclosarin (GF), VX and RVX in human urine were from 0.5 to 5 ng/mL. Recoveries of (40–80%) were obtained in the range of 10–300 ng/mL, with a linear response (R2 > 0.964, R > 0.982). The method is highly stable, even with DUS samples stored up to 5 months at room temperature before analysis. It was implemented in a sarin in vivo exposure experiment on mice, applied for the time course determination of isopropyl methylphosphonic acid (IMPA, sarin hydrolysis product) in mice urine. IMPA was detectable even with samples drawn 60 h after the mice’s (IN) exposure to 1 LD50 sarin. This method was also evaluated in a non-targeted screening for multiple potential CWA analogs (LC-Orbitrap HRMS analysis followed by automatic peak detection and library searches). The method developed here is applicable for rapid CWA casualty monitoring.

A-350 Myth Debunked: Same UrineAliquot is not Suitable for hCG Test AfterMicroscopic Analysis

Abstract Background It has been imparted in the laboratories that the suspending fluid, Lamina, from Iris IQ 200 Analyzer system is prone to interfere with urine human chorionic gonadotropin (hCG) test, especially on a small volume urine specimen. The myth is that the sample is diluted or contaminated by Lamina, which gives falsely negative result for hCG test. An add-on urine pregnancy test was ordered for a specimen, which was completed urinalysis and microscopic analysis. This is the only urine aliquot available, and it is approximately 2mL. This order raised many concerns among laboratory technicians regarding the accuracy of the result. However, no documents or articles were found to support the suspicions. In this study, A serial of experiments was completed to address this concern. Methods Known pregnant patients were confirmed their pregnancies with serum hCG tests prior to collecting the urine specimens. The experiment was conducted with 4 positive patient urines. A series of samples ran on DXI 800 system and the point-of-care (POC) Alere hCG combo cassettes. Then the same specimen performed urinalysis and microscopic analysis on Iris IQ 200 Analyzer. To evaluate the Lamina impact, the sample concentrations were verified with point-of-care (POC) Alere hCG combo cassettes and later DXI 800 system. Meantime, the significance of sample size was also evaluated in the study. Results The final concentrations of the samples were at <5, 5, 10, 15, 20, 25, 30, 40, 50, and 60 mIU/mL. The cutoff for Alere hCG combo cassettes was verified at 20 mIU/mL for urine, the assay gives the accurate result at and above that value. The results are consistent between immunoassay and POCT prior to and after microscopic analysis. Therefore, no significant interferences were identified with the desired sample volume, 3 mL, or short sample volume, 2mL. The volume usage by iQ 200 Analyzer was less than 500 uL. Conclusion In this study, Lamina doesn't cause any interference with hCG values using Alere hCG combo cassettes despite the short sample volume. However, variations between POC urinary hCG devices may contribute to different outcomes. Therefore, a separate validation is highly recommended if a different POC urinary hCG device is involved.

An accurate diagnostic approach for urothelial carcinomas based on novel dual methylated DNA markers in small-volume urine.

An accurate diagnostic approach for urothelial carcinomas based on novel dual methylated DNA markers in small-volume urine.

Generation of mesenchymal stromal cells from urine-derived iPSCs of pediatric brain tumor patients.

Human induced pluripotent stem cells (iPSCs) provide a virtually inexhaustible source of starting material for next generation cell therapies, offering new opportunities for regenerative medicine. Among different cell sources for the generation of iPSCs, urine cells are clinically relevant since these cells can be repeatedly obtained by non-invasive methods from patients of any age and health condition. These attributes encourage patients to participate in preclinical and clinical research. In particular, the use of urine-derived iPSC products is a convenient strategy for children with brain tumors, which are medically fragile patients. Here, we investigate the feasibility of using urine samples as a source of somatic cells to generate iPSC lines from pediatric patients with brain tumors (BT-iPSC). Urinary epithelial cells were isolated and reprogrammed using non-integrative Sendai virus vectors harboring the Yamanaka factors KLF4, OCT3/4, SOX2 and C-MYC. After reprogramming, BT-iPSC lines were subject to quality assessment and were compared to iPSCs obtained from urine samples of non-tumor pediatric patients (nonT-iPSC). We demonstrated that iPSCs can be successfully derived from a small volume of urine obtained from pediatric patients. Importantly, we showed that BT-iPSCs are equivalent to nonT-iPSCs in terms of morphology, pluripotency, and differentiation capacity into the three germ layers. In addition, both BT-iPSCs and nonT-iPSCs efficiently differentiated into functional mesenchymal stem/stromal cells (iMSC) with immunomodulatory properties. Therefore, this study provides an attractive approach to non-invasively generate personalized iMSC products intended for the treatment of children with brain tumors.

90Sr bioassay in small-volume urine by ICP-MS/MS with CO2 as the reaction gas

Further mitigation: CO2 is a better reaction gas than O2 to overcome isobaric/polyatomic interferences caused by Zr and Ge for 90S analysis by ICP-MS/MS. The method detection limit was 0.978 pg L−1 (5 Bq L−1) in 10 mL urine, with a reasonable throughput (∼6 h for 12 samples).

Urine stabilization and normalization strategies favor unbiased analysis of urinary EV content

Urine features an ideal source of non-invasive diagnostic markers. Some intrinsic and methodological issues still pose barriers to its full potential as liquid biopsy substrate. Unlike blood, urine concentration varies with nutrition, hydration and environmental factors. Urine is enriched with EVs from urinary-genital tract, while its conservation, purification and normalization can introduce bias in analysis of EV subsets in inter-and intra-individual comparisons. The present study evaluated the methods that decrease such biases such as appropriate and feasible urine storage, optimal single-step EV purification method for recovery of proteins and RNAs from small urine volumes and a normalization method for quantitative analysis of urine EV RNAs. Ultracentrifugation, chemical precipitation and immuno-affinity were used to isolate EVs from healthy donors’ urine that was stored frozen or at room temperature for up to 6 months. Multiple urine biochemical and EV parameters, including particle count and protein content, were compared across urine samples. To this purpose nanoparticle tracking analysis (NTA) and protein assessment by BCA, ELISA and WB assays were performed. These measurements were correlated with relative abundances of selected EV mRNAs and miRNAs assessed by RT-PCR and ranked for the ability to reflect and correct for EV content variations in longitudinal urine samples. All purification methods enabled recovery and downstream analysis of EVs from as few as 1 ml of urine. Our findings highlight long term stability of EV RNAs upon urine storage at RT as well as excellent correlation of EV content in urine with some routinely measured biochemical features, such as total urine protein and albumin, but not creatinine most conventionally used for urine normalization. Comparative evaluation of mRNA and miRNAs in EV isolates revealed specific RNAs, in particular RNY4 and small miRNA panel, levels of which well reflected the inter-sample EV variation and therefore useful as possible post-analytical normalizers of EV RNA content. We describe some realistic urine processing and normalization solutions for unbiased readout of EV biomarker studies and routine clinical sampling and diagnostics providing the input for design of larger validation studies employing urine EVs as biomarkers for particular conditions and diseases.

Comparisons of protocols for enhanced DNA profile quality from FTA®-deposited urine samples

Disputes over the identity of a urine sample donor have been reported, and urine authentication by genetic profiling has helped resolved the cases. However, since genotyping of urine is not always required, many drug-testing laboratories may face sample storage issues. Several studies have investigated the use of FTA® cards as a convenient tool for keeping specimen at room temperature for extended periods of time. However, generating complete STR profile from some FTA®-deposited urine samples remains challenging due to low levels of genetic material content, necessitating amendments to the laboratory’s standard protocols. This work therefore aims to evaluate the effects of two DNA template preparation methods, both employing FTA® cards as the storage medium, on the success rates of STR profiling from urine. Specimen from a female volunteer, representing a particularly low-yield sample, was employed. Aliquots of 1 and 2 mL were used as the starting material to evaluate DNA template preparation using the FTA® manufacturer’s protocol for disc purification against elution of DNA from the FTA® using Prepfiler™ Forensic DNA Extraction Kit. AmpFℓSTR™ Identifiler™ Plus PCR Amplification Kit was used to amplify the STR markers, and the PCR products were analysed using Applied Biosystems™ 3500xL Genetic Analyzer. The DNA profile qualities were examined in terms of number of loci detected and peak height balance. Comparisons with the profiles obtained from DNA isolated using QIAamp® DNA Micro Kit from 1 and 2 mL of the same batch of urine were also made. The optimised protocol was then tested on urine samples from three male volunteers. The results showed that the purification of FTA® punches according to the manufacturer’s protocol enabled full DNA profiles to be obtained from both 1 and 2 mL of urine from all samples tested, including male samples. In contrast, no DNA profile could be generated from the DNA eluted with the Prepfiler™ kit. When compared with the more conventional solid-phase DNA extraction method, the profiles generated from the FTA® punches exhibited similar reproducibility and quality to those from the template isolated by the QIAamp® Kit. This work further demonstrated the feasibility of FTA® cards as a tool for specimen storage and DNA template preparation from small volumes of urine for authentication by STR profiling. Full STR profiles could be generated from sample from both sexes without modification of the PCR conditions or injection time.

An optimized method for the isolation of urinary extracellular vesicles for molecular phenotyping: detection of biomarkers for radiation exposure

BackgroundUrinary extracellular vesicles (EVs) are a source of biomarkers with broad potential applications across clinical research, including monitoring radiation exposure. A key limitation to their implementation is minimal standardization in EV isolation and analytical methods. Further, most urinary EV isolation protocols necessitate large volumes of sample. This study aimed to compare and optimize isolation and analytical methods for EVs from small volumes of urine.Methods3 EV isolation methods were compared: ultracentrifugation, magnetic bead-based, and size-exclusion chromatography from 0.5 mL or 1 mL of rat and human urine. EV yield and mass spectrometry signals (Q-ToF and Triple Quad) were evaluated from each method. Metabolomic profiling was performed on EVs isolated from the urine of rats exposed to ionizing radiation 1-, 14-, 30- or 90-days post-exposure, and human urine from patients receiving thoracic radiotherapy for the treatment of lung cancer pre- and post-treatment.ResultsSize-exclusion chromatography is the preferred method for EV isolation from 0.5 mL of urine. Mass spectrometry-based metabolomic analyses of EV cargo identified biochemical changes induced by radiation, including altered nucleotide, folate, and lipid metabolism. We have provided standard operating procedures for implementation of these methods in other laboratories.ConclusionsWe demonstrate that EVs can be isolated from small volumes of urine and analytically investigated for their biochemical contents to detect radiation induced metabolomic changes. These findings lay a groundwork for future development of methods to monitor response to radiotherapy and can be extended to an array of molecular phenotyping studies aimed at characterizing EV cargo.

Simultaneous determination of 16 urinary metabolites of organophosphate flame retardants and organophosphate pesticides by solid phase extraction and ultra performance liquid chromatography coupled to tandem mass spectrometry

Organophosphate flame retardants (OPFRs) and organophosphate pesticides (OPPs), pertaining to organophosphate esters, are ubiquitous in environment and have been verified to pose noticeable risks to human health. To evaluate human exposures to OPFRs and OPPs, a fast and sensitive approach based on a solid phase extraction (SPE) followed by the ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) detection has been developed for the simultaneous analysis of multiple organophosphorus metabolites in urine. The method allows the identification and quantification of ten metabolites of the most common OPFRs and all six dialkylphosphates (DAPs) of OPPs concerning the population exposure characteristics. The method provided good linearities (R2 = 0.998–0.999), satisfactory method detection limits (MDLs) (0.030–1.129 ng/mL) and only needed a small volume (200 μL) of urine. Recovery rates ranged 73.4–127.1% at three spiking levels (2, 10 and 25 ng/mL urine), with both intra- and inter-day precision less than 14%. The good correlations for DAPs in a cross-validation test with a previous gas chromatography-mass spectrometry (GC-MS) method and a good inter-laboratory agreement for several OPFR metabolites in a standard reference material (SRM 3673) re-enforced the precision and validity of our method. Finally, the established method was successfully applied to analyze 16 organophosphorus metabolites in 35 Chinese children's urine samples. Overall, by validating the method's sensitivity, accuracy, precision, reproducibility, etc., data reliability and robustness were ensured; and the satisfactory pilot application on real urine samples demonstrated feasibility and acceptability of this method for being implemented in large population-based studies.

Detection of Biomarkers Associated with Acute Kidney Injury by a Gold Nanoparticle Based Colloidal Nano-Immunosensor by Fourier-Transform Infrared Spectroscopy with Principal Component Analysis

A colloidal nano-immunosensor based on gold nanoparticles covered in their surface with protein A and the immunoglobulin anti-kidney injury molecule-1 was prepared and tested in human urine to identify a biomarker that diagnose acute kidney injury. The urine was obtained from patients with cancer treated with cisplatin, diagnosed with acute kidney injury, and from healthy individuals. Fourier-transform infrared spectroscopy analysis showed important spectral differences between both types of groups, primarily for the intervals from 1750 to 1500 cm−1 and from 1000 to 891 cm−1 where the amide I group (C = O) and the glycosylated functional groups of this protein absorb. The principal component analysis applied to the vibrational signals obtained with the nano-immunosensor allows adequate discrimination of urine from patients with acute kidney injury from samples from healthy individuals. The simplicity of the proposed method, from the preparation of the nano-immunosensor to the spectral and chemometric analysis, may make this method an interesting alternative to practically diagnose acute kidney injury using a small volume of urine.

Label-Free, Novel Electrofluidic Capacitor Biosensor for Prostaglandin E2 Detection toward Early and Rapid Urinary Tract Infection Diagnosis.

Urine Prostaglandin E2 (PGE2) has been identified as an attractive diagnostic and prognostic biomarker for urinary tract infection (UTI). This work demonstrates the use of PGE2 as a biomarker for rapid and label-free testing for UTI. In this work, we have developed a novel electrofluidic capacitor-based biosensor that can used for home-based UTI management with high accuracy in less than 5 min for small volume urine samples (<60 μL). The PGE2 biosensor works on the principle of affinity capture using highly specific monoclonal PGE2 antibody and relies on non-faradaic electrical impedance spectroscopy (EIS) and Mott-Schottky (MS) for quantifying subtle variations in PGE2 levels expressed in human urine (pH 5-8). Dynamic light scattering experiments were performed to characterize surface charge properties and the impact of bulk interferents on the interfacial modulation of electrical properties due to binding and urine pH variations. Binding chemistry between the key elements of the immunosensor stack was validated using attenuated total reflectance-Fourier transform infrared spectroscopy and surface plasmon resonance studies. Linear calibration dose responses were obtained for PGE2 for both EIS and MS. The sensor reliably distinguished between UTI negative and UTI positive cases for both artificial (pH 5-8) and pooled human urine samples. The sensor was not found to cross-react with Prostaglandin D2, a structurally similar interferent, and other abundant urine interferents (urea and creatinine). Human subject studies confirmed the validity of the sensor for robust and accurate UTI diagnosis. This work can be extended to achieve easy, reliable, and rapid home-based UTI management, which can consequently help physicians with timely and appropriate administration of therapy to improve patient outcomes and treatment success.

Development, validation, and implementation of an UHPLC-MS/MS method for the quantitation of furosemide in infant urine samples.

Furosemide is a diuretic drug used to increase urine flow in order to reduce the amount of salt and water in the body. It is commonly utilized to treat preterm infants with chronic lung disease of prematurity. There is a need for a simple and reliable quantitation of furosemide in human urine. We have developed and validated an ultra-high performance liquid chromatography-tandem mass spectrometry method for furosemide quantitation in human urine with an assay range of 0.100-50.0μg/ml. Sample preparation involved solid-phase extraction with 10μl of urine. Intra-day accuracies and precisions for the quality control samples were 94.5-106 and 1.86-10.2%, respectively, while inter-day accuracies and precision were 99.2-102 and 3.38-7.41%, respectively. Recovery for furosemide had an average of 23.8%, with an average matrix effect of 101%. Furosemide was stable in human urine under the assay conditions. Stability for furosemide was shown at 1week (room temperature, 4, -20 and -78°C), 6months (-78°C), and through three freeze-thaw cycles. This robust assay demonstrates accurate and precise quantitation of furosemide in a small volume (10μl) of human urine. It is currently being implemented in an ongoing pediatric clinical study.

An eco-friendly solvent-free reaction based on peptide probes: design an extraction-free method for analysis of acrylamide under microliter volume.

Acrylamide is a group 2A carcinogen and potential endocrine disruptor that can enter the ecosystem by various routes and has recently become a dangerous pollutant. This widely used chemical can enter the human body via air inhalation, food or water consumption, or skin contact. In this study, we developed a peptide probe for the detection of acrylamide by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) after its micro-tagging with a peptide. Direct detection of acrylamide by MALDI-TOF MS is not feasible due to its poor ionization in the MALDI interface, which hinders its analysis by the technique. After microwave irradiation for 2min, the formed acrylamide-peptide derivative was detected easily by MALDI-TOF MS without the need for extraction procedures. The procedure does not involve organic solvents and a water-soluble peptide that allows detection of acrylamide in small sample volumes with a limit of detection (LOD) of 0.05ng/μL. The relative standard deviation (RSD) and relative error (RE) of the measurements were < 6.7% for intra- and inter-day assays. Gel-washing solutions from a polyacrylamide gel experiment were used as a model to study the efficiency of the developed method. Finally, we used the proposed method for the detection of free acrylamide in small volumes of lung epithelial cells (a model to test the air inhalation of acrylamide under a tiny volume of sample) and human urine. The developed method will enable rapid acrylamide detection in environmental and biological samples via a green approach based on microwave-assisted derivatization in water alongside the use of a less toxic derivatization reagent, reusable target plate, and miniaturization protocols.

Rapid analysis of 237Np and Pu isotopes in small volume urine by SF-ICP-MS and ICP-MS/MS

Internal contamination with alpha-particle emitting actinides, such as 237Np, 239Pu, 240Pu, is likely to bring a large amount of dose to the tissues of persons even if the intake amount is small. To provide timely information for prompt decision-making in radiation emergency therapy, we developed a simple and rapid method for urinary bioassay to determine ultra-trace 237Np and Pu isotopes using SF-ICP-MS and ICP-MS/MS. To avoid polyatomic interferences and tailing effects from U, 237Np and Pu isotopes were collected after removing U effectively using a simple single chromatographic column packed with 2 mL AG MP-1M anion exchange resin, exhibiting a high decontamination factor of 108 for 238U. The overall chemical fractionation between 237Np and 242Pu for the whole analytical procedure was 0.974 ± 0.064 (k = 2), allowing us to measure 237Np and Pu isotopes using 242Pu as a yield tracer with yields of 76 ± 5%. Using ICP-MS/MS with low background provided the method detection limits for 237Np, 239Pu, 240Pu, and 241Pu of 0.025, 0.025, 0.015, and 0.020 fg mL−1, respectively, for 20 mL of urine sample. Those were comparable to detection limits of SF-ICP-MS with high sensitivity. Subsequently, three urine reference materials with Pu spike, provided by the Association for the PROmotion of Quality COntrol in RADiotoxicological Analysis (PROCORAD), France, were analyzed by the developed method and the conventional alpha spectrometry technique for validation. Finally, the developed method was successfully employed to measure the contamination level of 237Np, 239Pu, 240Pu, and 241Pu in urine samples collected during decorporation therapy using DTPA, after a Pu inhalation exposure accident in Japan. The high throughput (9 h for 12 samples), simplicity, low cost, and high sensitivity of the method will allow greater numbers of related laboratories to be involved in screening activities for unexpected actinide exposure, such as in the case of a large scale radiological disaster.

Reliability of a dried urine test for comprehensive assessment of urine hormones and metabolites

BackgroundMass spectrometry allows for analysis of multiple hormone and organic acid metabolites from small urine volumes; however, to assess the full extent of daily hormone production, 24-h urine collections are usually required. The aims of this study were, first, to confirm that mass spectrometric analysis of an array of hormones and organic acids would yield similar results in both liquid and dried urine, and, second, to determine if collection of four dried spot urine samples could be substituted for a 24-h collection when measuring reproductive hormones.MethodsTwo study populations were included in this prospective observational study. Twenty individuals collected both a spot liquid urine and dried urine on filter paper to analyze eight organic acids. A second group of 26 individuals collected both a 24-h urine and four dried spot urines during waking hours throughout the same day for evaluation of 17 reproductive hormones and metabolites; data from 18 of these individuals were available to compare liquid versus dried urine results. Dried urine was extracted, hydrolyzed, and derivatized before analysis by mass spectrometry; all analytes from dried urine were normalized to urine creatinine.ResultsReproductive hormone results from dried and liquid urine were in excellent agreement with intraclass correlation coefficients (ICCs) greater than 0.90; comparison of dried to liquid urine for organic acids showed good to excellent agreement (ICC range: 0.75 to 0.99). Comparison between the 4-spot urine collection and 24-h urine collection methods showed excellent agreement (ICC > 0.9) for 14 of the 17 urine metabolites and good agreement for the others (ICC 0.78 to 0.85) with no systematic differences between the two methods of collection.ConclusionsThe burden of urine collection can be reduced using collection of four spot dried urines on filter paper without compromising comparability with hormone results from a 24-h urine collection. A large number of urine analytes can be assessed from the dried urine with similar results to those from liquid urine. Given the ease of sample handling, this 4-spot dried urine assay would be useful for both clinical assessment of patients and for large epidemiologic studies.

Clinical Profiles and Factors Associated with a Low Sodium Intake in the Population: An Analysis of the Swiss Survey on Salt.

As a part of the salt controversy, it has been suggested that people with a low sodium intake have an increased risk of cardiovascular events. However, there is no clear explanation for this increased risk. We examined the socio-demographic, clinical profile, and behavioral factors associated with a low sodium intake in the Swiss subjects who participated in the Swiss Survey on Salt. Only 13.3% of the Swiss population eat less than 5 g of salt daily and among them 78.2% are women. Subjects with a low sodium intake eat and drink less as reflected by lower intakes of proteins, potassium, and calcium and a smaller urine volume. In addition, a low blood pressure, a normal body mass index, a low prevalence of obesity, a low serum uric acid, and less alcohol and cigarette consumption characterized this group, suggesting a rather low cardiovascular risk profile. Being single and doing most of the cooking at home are associated with a low intake of sodium, as well as a less frequent consumption of meat and fish when eating less than 5 g salt per day. However, the awareness of the effects of salt on health and cardiovascular risk, health concerns, and physical activity are similar in subjects eating more or less salt. In conclusion, we could not evidence clinical or behavioral factors that could significantly increase the risk of developing cardiovascular events in low salt eaters.

A new procedure for the determination of 21 macro- and trace elements in human fetal urine using an inductively coupled plasma mass spectrometry with dynamic reaction cell (ICP-DRC-MS) equipped with a micro-flow nebulizer

The procedure for determination of 21 macro- and trace elements – Li, Na, Mg, Al, K, Ca, V, Cr, Mn, Fe, Co, Cu, Zn, Se, Sr, As, Cd, Sb, Ba, Pb and U – in human fetal urine by inductively coupled plasma mass spectrometry (ICP-MS) was developed and validated. The application of a micronebulizer and a dynamic reaction cell (DRC) allowed to perform a full analysis of small volumes (200 μL) of urine collected from human fetuses without the need for sample digestion with closed microwave systems. The procedure and ICP-MS instrument was thoroughly optimized in order to reliably determine both macroelements and ultra-trace concentrations of elements. The internal standard method (Ge, Rh and Tb) was applied in order to encompass signal drift and non-spectral interferences. The rules of metrology were used in order to ensure the quality of the results: (1) the procedure was validated, (2) the uncertainty of the measurement results was estimated and (3) the traceability of the measurement result was established by using the certified reference material with matching matrix (Seronorm Trace Elements Urine L-1). Also, the analyte addition method to the artificial urine was employed for additional confirmation of trueness of the procedure. The selected parameters of the procedure were as follows: (a) limits of detection – (0.00023–53 μg L−1) for U and Ca, respectively, (b) recoveries of the reference value – 81%–136% for Mn and Cd, respectively (c) linearity expressed as R – greater than 0.999, and (d) expanded relative uncertainties (k = 2) – 13%–66% for Sr and Cd, respectively. The developed and validated procedure was applied to 58 samples of urine collected from human fetuses. The samples were diluted with nitric acid and analyzed without further treatment. The procedure allowed to reliably determine both macro- and trace elements in very low volume of sample in a single analytical run.

A pilot study to compare urinary protein of fractional urine collected over 24h period versus traditional 24h urine collection method

24 hours traditional urine collection method is cumbersome and tedious process with less patient compliance. Our study proposes a novel fractional urine collection method for protein estimation that would entail collection of small volumes of urine each time patient voids Over 24h Period. Our aim was to compare urine protein in proposed method versus traditional 24hour sampling method. Cross-sectional, prospective study was a collaborative work between Departments of Nephrology and Biochemistry in our tertiary care hospital. Volunteers (48) and Chronic Kidney disease subjects (52) were recruited. Out of which, 76 subjects were selected. Novel method was compared with conventional method in each of these groups, before and after centrifugation and with and without preservative. Intraclass correlation coefficient (ICC) and Bland Altman analysis was used to evaluate for agreement between the two methods. All the values were combined without categorization into subgroups. Fractional urine collection method (before centrifugation) without preservative (F1) and with preservative (F2) was compared to conventional method, ICC was 0.93 (C.I, 0.9 to 0.95) and 0.96 (C.I, 0.94 to 0.97) respectively. After centrifugation, fractional urine collection method for F1and F2 versus conventional method, ICC was 0.95 (C.I, 0.92 to 0.97) and 0.91 (C.I, 0.86 to 0.94) respectively. The novel collection method is comparable and reliable as traditional 24-hour method. The use of thymol as preservative leads to negative interference and centrifugation is mandatory before urine analysis. With proper sampling, urine protein by novel method has a good agreement with the conventional technique.