SEARCH · Search PubMed

Results for “Sample pretreatment”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

SourceResearch connectionFrom yearThrough year

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 451 records · Page 25Linked to original sources

PubMed2001

Determination of the human salivary peptides histatins 1, 3, 5 and statherin by high-performance liquid chromatography and by diode-array detection.

A reversed-phase high-performance liquid chromatography (HPLC) method with diode-array detection for the quantification of several human salivary peptides is described. Sample pretreatment consisted of the acidification of whole saliva by phosphate buffer. This treatment produced precipitation of mucins, alpha-amylases and other high-molecular-mass salivary proteins, simultaneous inhibition of intrinsic protease activities and reduction of sample viscosity. Direct HPLC analysis by diode-array detection of the resulting acidic sample allowed one to quantify histatin 1, histatin 3, histatin 5, statherin, as well as uric acid, in normal subjects. Moreover, the groups of peaks pertaining to proline-rich proteins and cystatins were tentatively identified. The method can be useful in assessing the concentration of salivary peptides from normal subjects and from patients suffering oral and/or periodontal diseases.

Acids↗

PubMed2003

Liquid chromatographic analysis of the cis(Z)- and trans(E)-isomers of clopenthixol in human plasma using a novel solid phase extraction procedure.

A simple and highly sensitive high-performance liquid chromatographic (HPLC) method for the simultaneous determination of cis(Z)-clopenthixol and trans(E)-clopenthixol in human plasma has been developed. The chromatographic analysis was carried out isocratically on a reversed-phase column (C(8) 150 x 4.6 mm I.D., 5 microm) using a mixture of 25 mM phosphate buffer and acetonitrile (65:35 v/v, pH* 3.0) as the mobile phase, and ultraviolet detection at 230 nm. Plasma sample pretreatment was accomplished by means of an original solid-phase extraction (SPE) procedure carried out on cyanopropyl cartridges, with a high extraction yield and good selectivity. Under the optimum conditions, calibration graphs of spiked human plasma samples were obtained over the concentration ranges 1-300 ng ml(-1) for cis(Z)-clopenthixol and 1-200 ng ml(-1) for trans(E)-clopenthixol. The limit of detection (LOD) was 0.3 ng ml(-1) for both cis(Z)- and trans(E)-isomers of clopenthixol. The method was successfully applied to the determination of cis(Z)-clopenthixol and trans(E)-clopenthixol in plasma samples of schizophrenic patients undergoing therapy with zuclopenthixol.

Antipsychotic Agents↗

PubMed2000

Field cryofocussing hydride generation applied to the simultaneous multi-elemental determination of alkyl-metal(loid) species in natural waters using ICP-MS detection.

Two hydride generation manifold systems, utilizing flow injection and cryotrapping techniques for alkyl-metal(loid) speciation analysis in natural waters, are described in this paper. They provide shipboard capacity for simultaneous derivatization of analytes with NaBH4 and cryotrapping of the generated products in a field packed column at -196 degrees C. The first system is a large-volume hydride generator, using a reagent-injection flow technique as a flow batch type, that has been fully optimized and applied to the simultaneous detection of alkylated species in estuarine waters. The technique permits the analysis of a large volume sample (0.5-11) at a sampling rate of 3 h-1. The second is an online continuous flow hydride generator. A sampling rate of 3-12 h-1 can be achieved with samples of 0.1-0.51. In addition, shipboard operation eliminates major problems related to sample pretreatment, transport and storage. Ultra-trace multi-element determination is finally performed in the laboratory by cryogenic GC hyphenated with ICP-MS. Routine detection limits of 0.5-10 pg (as metal) for 0.51 water samples were achieved for the selected alkyl-metal(loid) species of arsenic, germanium, mercury and tin. Concentrations of various species, obtained from water samples taken from the Rhine estuary, are also presented. These species include alkylated arsenic compounds, other than methyl derivatives, that have been tentatively identified and are reported here for the first time.

Chemistry Techniques, Analytical↗

PubMed2001

The potential of 19F NMR spectroscopy for rapid screening of cell cultures for models of mammalian drug metabolism.

The use of microbial cultures as a complementary model for mammalian drug metabolism has been well established previously. Here is a preliminary investigation into the potential of 19F NMR spectroscopy as a rapid screening tool to quantify the biotransformations of fluorine-containing model drugs. Biotransformations of three model drugs in 48 taxonomically diverse organisms were measured by acquiring 19F NMR spectra at 376 MHz. The presence of fluorine in the molecules allowed rapid, simultaneous detection of over 20 biotransformation products without sample pretreatment, chromatography, mass spectrometric techniques or the use of radiolabelled substrates. The detection limit at 376 MHz using 5 mm NMR tubes was ca. 0.3 microg ml(-1) using a typical analysis time of 20 min per sample. With the recent advent of flow injection NMR technology, analysis time of 5 min could be achieved with less sample. This approach may be used to develop fast small-scale microbial screens for the biosynthesis of metabolite standards and production of novel drug analogues, whilst also having a role in reducing animal experiments needed to identify animal and human metabolites of fluorinated xenobiotics.

Animals↗

PubMed1993

Diagnosis and management of extrauterine pregnancies.

This study was based on 16 women provisionally diagnosed as having extrauterine pregnancies. Of these, 13 (81.3%) were confirmed as positive at operation. Patients were managed according to 1 of 3 regimens; 1) methotrexate (n = 4), 2) methotrexate followed by surgery (n = 3) and 3) surgery (n = 6). Serial blood samples, collected before and after treatment, were analyzed for ovarian (oestradiol, E2; progesterone, P4) uterine (placental protein 14, PP14) and placental markers (chorionic gonadotrophin, HCG; pregnancy-associated plasma protein-A (PAPP-A). Of the pretreatment samples, only 30.4% and 41.7% were depressed for PP14 and HCG, respectively. By contrast, the diagnostic value of PAPP-A (77.8%) and P4 (87.5%) was greater. Biochemical monitoring of treatment was best achieved with trophoblastic derived antigens (HCG), whereas antigens of maternal origin demonstrated widely varied responses. This study established the effectiveness of chemotherapy for treatment of tubal pregnancies as an alternative to surgery, but if a biochemical marker is required, the marker of choice is HCG.

Abortion, Therapeutic↗

PubMed1992

A fully automated enzyme immunoassay for the measurement of cortisol in biological fluids.

This paper describes a fully automated assay on the Serono SR1 for the measurement of cortisol in serum, heparinised plasma and urine. The assay incorporates a specific polyclonal antibody to cortisol and cortisol conjugated to alkaline phosphatase as a label. Following immunoincubation, bound and free labelled cortisol are separated by magnetic sedimentation of the antibody complex. Phenolphthalein is liberated by the enzymatic hydrolysis of the substrate (phenolphthalein monophosphate) and the absorbance generated is measured as the assay end-point. All processing steps are performed by the SR1. The assay has good analytical performance with respect to precision (within and between run CVs less than 10 and 11.5% respectively), detection limit (less than 5 nmol/l) and recovery of added cortisol (100 +/- 10%). The assay agrees well with cortisol concentrations determined by ID-MS and by established immunoassays (r > 0.96). Reference ranges of normal urine samples (pretreated by solvent extraction) are in good agreement with accepted values. The study demonstrates that the SR1 Cortisol assay on the SR1 analyzer is suitable for the routine determination of cortisol in serum, heparinised plasma and urine samples.

Adult↗

PubMed2002

Determination of oxalate in urine by zone electrophoresis on a chip with conductivity detection.

The use of a poly(methylmethacrylate) capillary electrophoresis chip, provided with a high sample load capacity separation system (a 8500 nL separation channel coupled to a 500 nL sample injection channel) and a pair of on-chip conductivity detectors, for zone electrophoresis (ZE) determination of oxalate in urine was studied. Hydrodynamic and electroosmotic flows of the solution in the separation compartment of the chip were suppressed and electrophoresis was a dominant transport process in the separations performed on the chip. A low pH of the carrier electrolyte (4.0) provided an adequate selectivity in the separation of oxalate from anionic urine constituents and, at the same time, also a sufficient sensitivity in its conductivity detection. Under our working conditions, this anion could be detected at a 8 x 10(-8) mol/L concentration also in samples containing chloride (a major anionic constituent of urine) at 3.5 x 10(-3) mol/L concentrations. Such a favorable analyte/matrix concentration ratio (in part, attributable to a transient isotachophoresis stacking in the initial phase of the separation) made possible accurate and reproducible (typically, 2-5% relative standard deviation (RSD) values of the peak areas of the analyte in dependence on its concentration in the sample) determination of oxalate in 500 nL volumes of 20-100-fold diluted urine samples. Short analysis times (about 280 s), no sample pretreatment (not considering urine dilution) and reproducible migration times of this analyte (0.5-1.0% RSD values) were characteristic for ZE on the chip. This work indicates general potentialities of the present chip design in rapid ZE analysis of samples containing the analyte(s) at high ionic matrix/analyte concentration ratios.

Electric Conductivity↗

PubMed2001

Advances in sample preparation in electromigration, chromatographic and mass spectrometric separation methods.

The quality of sample preparation is a key factor in determining the success of analysis. While analysis of pharmaceutically important compounds in biological matrixes has driven forward the development of sample clean-up procedures in last 20 years, today's chemists face an additional challenge: sample preparation and analysis of complex biochemical samples for characterization of genotypic or phenotypic information contained in DNA and proteins. This review focuses on various sample pretreatment methods designed to meet the requirements for the analysis of biopolymers and small drugs in complex matrices. We discuss the advances in development of solid-phase extraction (SPE) sorbents, on-line SPE, membrane-based sample preparation, and sample clean-up of biopolymers prior to their analysis by mass spectrometry.

Chromatography↗

PubMed1999

Direct determination of small cations in proteinaceous samples using a flow injection-capillary electrophoresis system.

A method is described for the direct determination of small inorganic cations in samples containing large amounts of proteins, such as milk or blood plasma. The method is based on electrokinetic injection in a flow injection analysis-capillary electrophoresis (CE) system. The selected CE-electrolyte, containing 5 mM 4-aminopyridine and 7 microM cetyltrimethylammonium bromide at pH 4.5, prevents detrimental protein adsorption on the capillary walls. Therefore, no sample pretreatment, except for dilution, is required. Up to 30 repeated injections in one electrophoretic run can be performed, yielding RSD values of the migration time of less than 1 and 2.5% (n=30) for milk and blood plasma samples, respectively.

Blood Proteins↗

PubMed2000

Pressurized hot water extraction coupled on-line with LC-GC: determination of polyaromatic hydrocarbons in sediment.

Pressurized hot water extraction (PHWE) was directly combined with a LC-GC system for the determination of polyaromatic hydrocarbons (PAHs) in sediment. The sediment sample was first extracted with pressurized hot water, and the analytes were adsorbed into a solid-phase trap. The trap also functioned as a LC column, which removed most of the interfering matrix components. The 780-microL LC fraction containing the analytes was directly transferred to the GC using an on-column interface. The whole PHWE-LC-GC analysis took place in a closed system, and no sample pretreatment was required. The sensitivity of the method was excellent due to the efficient concentration in the LC-GC system. Sensitivity was approximately 800 times better than in traditional systems. In addition, only a small amount of sample (10 mg) was required for the analysis. The PHWE-LC-GC method proved to be linear in the concentration range of 0.01-2 microg/g, the limits of quantification were below 0.01 microg/g for all the analytes, and the relative standard deviations were between 3 and 28%. LC cleanup and the improved sensitivity made detection with FID sufficient for the determination of analytes. The results were comparable to those obtained in an interlaboratory comparison study as well as to the results obtained with off-line SFE-GC-MS.

Chromatography, Gas↗

PubMed1991

Spurious serologic test results in patients receiving infusions of intravenous immune gammaglobulin.

Intravenous immune gammaglobulin (IVIgG) has a definite role in the modern armamentarium of therapies for immunocompromised patients. Intravenous IgG is manufactured from the plasma of a large number of donors and contains antibodies against numerous infectious agents. Patients who receive infusions of IVIgG may test false-positive for a variety of viral, bacterial, and other infectious diseases when serologic determinations are performed on their blood samples. It is important that laboratories performing serologic testing be aware that patients have been infused with IVIgG, so that spurious laboratory results can be identified. Serologic testing on implicated lots of IVIgG and on the patient's pretreatment sample may aid in correctly identifying a spurious serologic result due to therapy with IVIgG.

Adolescent↗

PubMed1998

Microparticle-enhanced nephelometric immunoassay of lysozyme in milk and other human body fluids.

Quantitation of lysozyme in human milk was performed by a microparticle-enhanced nephelometric immunoassay based on the measurement of the light scattered during the competitive immunoagglutination of a microparticle-lysozyme conjugate with an anti-lysozyme antiserum. This immunoassay has a detection limit of 8 microg/L of reaction mixture and can be performed using diluted milk (1:6000, in reaction mixture), excluding sample pretreatment. Human milk lysozyme can be quantified over the concentration range 0.09-1.50 g/L, with within- and between-run coefficients of variation <5%. Changes in the lysozyme concentration of human milk during lactation were determined in 636 samples. Lysozyme concentrations (mean +/- SE) decreased from colostrum (0.36 +/- 0.02 g/L) to transitional milk (0.30 +/- 0.01 g/L) and mature milk during days 15-42 (0.30 +/- 0.01 g/L), then increased in the mature milk during days 43-56 (0.35 +/- 0.01 g/L) and especially during days 57-84 (0.83 +/- 0.05 g/L). The proportion of lysozyme contributing to total protein was found to rise during lactation and was as follows: colostrum (1.7%), transitional milk (2.3%), and mature milk from days 15-28 (2.7%), days 29-42 (3.1%), days 43-56 (3.8%), and days 57-84 (7.3%). The assay developed for milk was also suitable for the determination of lysozyme in other human body fluids.

Adolescent↗

PubMed2001

Assessment of murine bladder permeability with fluorescein: validation with cyclophosphamide and protamine.

OBJECTIVES: Bladder hyperpermeability should result in elevated blood levels of intravesically administered agents. Reabsorption from a hyperpermeable bladder should result in prolonged urinary excretion of an agent after parenteral administration. To test these hypotheses, urinary clearance and plasma levels of sodium fluorescein (NaF) were measured in mice before and during cyclophosphamide (CYP) and protamine-induced hemorrhagic cystitis. METHODS: To measure the plasma uptake of NaF from the bladder, 10 mg/mL NaF was instilled, either by catheter or retrograde urethral infusion, 15 minutes before retro-orbital or ventricular sampling. The plasma levels were measured 24 hours and 14 days after exposure to CYP 300 mg/kg or 15 minutes after instillation of protamine 10 mg/mL. Hourly urine concentrations were measured immediately after intraperitoneal administration of 10 mg/kg NaF. Pretreatment samples were compared with those obtained 24 hours after intraperitoneal administration of 300 mg/kg CYP. RESULTS: Urinary NaF excretion was delayed in CYP-exposed mice. A bi-exponential model provided an appropriate fit of the data, both before and after CYP administration. The plasma levels of NaF were significantly elevated at 24 hours and 14 days after CYP exposure when sampled by ventricular nick or retro-orbitally. The median concentration of fluorescein in the protamine-treated mice was significantly higher than in the control mice. CONCLUSIONS: Fluorescein can be used to measure alterations in bladder permeability after bladder mucosal injury in mice. Urinary excretion of NaF is a bi-exponential process that is delayed after bladder mucosal injury, presumably because of increased mucosal permeability and resorption from the urine into the bloodstream.

Analysis of Variance↗

PubMed2026

Development and validation of an LC-MS/MS method for the quantification of the KRASG12C inhibitor divarasib.

Divarasib is a newly developed covalent KRASG12C inhibitor, currently under clinical investigation in a phase 3 trial in patients with non-small cell lung cancer (NSCLC). At the moment, very limited pharmacokinetic data are publicly known. However, obtaining more insight into the pharmacokinetic properties of divarasib is important, since this may provide a better understanding of its efficacy and safety risks. Pre-clinical studies have been performed in mouse models to evaluate the effect of drug transporters and drug-metabolizing enzymes on the plasma exposure and tissue distribution of divarasib. Therefore, a reliable quantification method is required. To our knowledge, no bioanalytical assay of divarasib has been published yet. Therefore, in this study we developed and validated an assay to quantify divarasib in human plasma and in eight different mouse-related matrices, and partially in mouse plasma, using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The method was initially evaluated over a concentration range of 1-10,000 nM. However, due to carry-over observed at 10,000 nM, the validated calibration range was established at 1-2000 nM, with matrix-dependent LLOQs of 1-10 nM. Erlotinib was used as an internal standard and acetonitrile was utilized to perform protein precipitation as sample pretreatment. Divarasib demonstrated stability in human plasma and in mouse plasma and tissue homogenates under various experimental conditions. A pilot in vivo study showed the applicability of our validated LC-MS/MS method. Ongoing clinical trials may collect plasma samples, and this developed method enables quantification of divarasib in both mouse and human plasma samples.

Animals↗

PubMed1998

High-performance liquid chromatographic analysis of the investigational anticancer drug 9-aminocamptothecin, as the lactone form and as the total of the lactone and the hydroxycarboxylate forms, in micro-volumes of human plasma.

A high performance liquid chromatographic (HPLC) assay is described for the determination of the investigational anticancer drug 9 aminocamptothecin (9-AC) as the lactone form (9AC(lac)) and as the total of the lactone and hydroxycarboxylate forms (9AC-(tot)), in micro volumes of plasma. The analytical methodology reported here involves a protein precipitation step with cold methanol (-30 degrees C) as sample pretreatment procedure. The methanolic extract is used for the determination of 9AC-(tot). The intact (active) lactone form of 9-AC is separated from the hydroxycarboxylate form in the methanolic plasma extract by solid phase extraction within 48 h after sampling and deproteination. After evaporation to dryness (nitrogen, 40 degrees C) the extract can be stored at -70 degrees C for at least 3 weeks. The drug is analysed by reversed-phase liquid chromatography on a Zorbax SB RP-18 column, using methanol-water eluent (pH 2.2) and fluorescence detection. The presented assay is linear over a concentration range 0.2-100 ng.ml-1 with a detection limit and a limit of quantitation of 0.05 and 0.2 ng.ml-1, respectively, for both 9-AC(tot) and 9-AC(lac) using a 100 ml plasma sample. The proposed method has been implemented in a phase I clinical trial for pharmacokinetic evaluation of this potential new drug.

Aged↗

PubMed2005

Direct analysis of pharmaceutical drug formulations using ion mobility spectrometry/quadrupole-time-of-flight mass spectrometry combined with desorption electrospray ionization.

A novel approach to the rapid analysis of pharmaceutical drug formulations using hyphenated ion mobility spectrometry (IMS) and time-of-flight mass spectrometry (ToF-MS) that requires no sample pretreatment or chromatographic separation is described. A modified quadrupole time-of-flight mass spectrometer containing an ion mobility drift cell was used for gas-phase electrophoretic separation of ions prior to ToF-MS detection. The generation of sample ions directly from tablets and cream formulations was effected by desorption electrospray ionization (DESI) using a modified electrospray ion source. The analysis of a range of over-the-counter and prescription tablet formulations is described, including histamine H2 receptor antagonist (ranitidine), analgesic (paracetamol), opiate (codeine), and aromatase inhibitor anticancer (anastrozole) drugs. The successful determination of active drugs from soft formulations, such as an antiseptic cream (chlorhexidine) and a nicotine-containing skin patch, is also presented. Limits of detection for the active drugs using the DESI/IMS/ToF-MS method fell within the high-picomole to nanomole range. In all cases, the use of ion mobility drift tube separation showed increased selectivity for active drug responses (present as low as 0.14% w/w) over excipient responses such as poly(ethylene glycol). Tandem mass spectrometric analysis of precursor ions separated by IMS allowed positive confirmation of active drugs with little loss of ion mobility efficiency. The ability to analyze hard or soft pharmaceutical formulations directly by DESI combined with ion mobility spectrometry/mass spectrometry in approximately 2 min demonstrates the potential applicability of this novel method to pharmaceutical screening of low-molecular-weight drug formulations with high selectivity over the formulation vehicle.

Chemistry, Pharmaceutical↗

PubMed2004

Epstein-Barr virus DNA measured in nasopharyngeal brushings in patients with nasopharyngeal carcinoma: pilot study.

OBJECTIVE: We measured the amount of tumour-derived Epstein-Barr virus (EBV) deoxyribonucleic acid (DNA) in the nasal brushings of nasopharyngeal carcinoma (NPC) patients to determine the correlation with tumour load and response to treatment. MATERIALS AND METHODS: Twenty-eight patients with NPC were included in the study. Baseline measurements of EBV from nasopharyngeal brushings were obtained from 26 patients prior to treatment. A follow-up sample was available from 11 of these patients post-treatment and from 2 additional patients who did not have a baseline sample. Quantitative real-time polymerase chain reaction (PCR) using SYBR Green I fluorescent dye was used to detect the EBV DNA copy number. RESULTS: Nasopharyngeal brush biopsies showed a high copy number of EBV DNA in most of the pretreatment samples (median 9714 copies/mL). The highest copy number detected was 14536944 copies/mL in one sample. In the post-treatment follow-up samples, the copy number was significantly lower (median 6 copies/mL). CONCLUSIONS: We have demonstrated that EBV DNA can be detected in the brush biopsies from NPC patients using quantitative real-time PCR. These pilot data suggest that nasopharyngeal brushings with PCR detection of EBV may be an effective tool for determining local tumour response. The potential of this technique as an NPC tumour marker for post-treatment follow-up is being validated with larger patient numbers.

Adult↗

PubMed2026

Circulating Tumor DNA Profiling Defines Risk Classification in Patients With Ewing Sarcoma: A Report From the Children's Oncology Group and the LEOPARD Study.

PURPOSE: Identification of discrete risk groups remains a high priority for patients with Ewing sarcoma (EWS). We sought to prospectively validate circulating tumor DNA (ctDNA) as a prognostic factor and develop clinical-molecular risk groups. METHODS: We conducted a prospective investigator-initiated biology study for patients with localized EWS (LEOPARD) and embedded ctDNA analysis into the North American frontline metastatic study AEWS1221. Eligible patients were younger than 50 years with newly diagnosed EWS. All patients provided a baseline blood sample for analysis, which was subjected to ultralow-pass whole-genome sequencing and hybrid capture panel sequencing for ctDNA quantification, fusion detection, and characterization of STAG2 and TP53 alterations. Serial ctDNA sequencing was conducted on a subset of patients in each study. We tested for associations between ctDNA burden and secondary genomic events, and clinical features and outcomes. RESULTS: One hundred forty patients with localized disease and 255 with metastatic disease provided evaluable pretreatment samples for ctDNA analysis. Elevated baseline ctDNA was associated with stage, tumor size, primary site, indeterminate pulmonary nodules, and metastatic pattern. Elevated pretreatment ctDNA burden was associated with inferior outcomes in patients with localized (n = 140, hazard ratio [HR] = 2.36, P = .032) and metastatic disease (n = 255, HR = 2.15, P = .001). Patients with metastatic disease and TP53 variants and/or persistent on-therapy ctDNA had dismal outcomes. Patients with localized disease, low ctDNA, small tumors, and favorable genomics had no events and constitute a novel low-risk group. Among patients with metastatic disease, those with lung-only disease, low ctDNA, and favorable genomics represent an intermediate-risk group. CONCLUSION: This study prospectively validates pretreatment ctDNA burden as prognostic in EWS. Risk groups that integrate ctDNA burden with clinical-molecular features differentiate patients with low-, intermediate-, and high-risk disease.

Journal Article↗