An LC-MS/MS method for creatine and creatinine analysis in paraquat-intoxicated patients

Abstract

A sudden increase in serum creatinine after paraquat intoxication has been reported in several clinical studies. However, this dramatic change of creatinine may be possibly due to an interconversion of creatine-creatinine in relation to paraquat toxicity. In order to investigate the creatine-creatinine relationship, a liquid chromatography tandem mass spectrometry in combination with electrospray ionization was developed and validated for simultaneous determination of creatine and creatinine in the serum. The chromatographic separation was achieved on a Gemini® C₆-Phenyl column with a gradient elution consisting of 0.1% formic acid in ultrapure water and methanol as the mobile phase. The method yielded suitable levels of specificity and selectivity, and calibration curves of creatine and creatinine in serum were linear over the concentration range of 0.5–200?µg mL^?1. The limit of quantification of both compounds was 0.5?µg mL^?1, and the method was accurate within the recovery range of 96.23–102.75%, indicating the robustness of the method. The method was successfully applied to toxicological samples from paraquat-intoxicated patients, and the concentrations of creatine and creatinine were quantified. High creatine concentrations in serum samples were observed which may lead to high serum creatinine despite normal kidney function as creatine is converted to creatinine in proportion to its concentration.     

Parental phenols exposure and spontaneous abortion in Chinese population residing in the middle and lower reaches of the Yangtze River

Widespread use of phenols has led to ubiquitous exposure to phenols. In experimental animals, phenols increased resorptions, reduced live litter size and fetal body weights. However, there are limited epidemiological evidences of the relationships between exposure to phenols and pregnancy outcomes. We evaluated the associations between parental urinary levels of various phenols and spontaneous abortion in a Chinese population residing in the middle and lower reaches of the Yangtze River. A case-control study was conducted that included 70 case couples with medically unexplained spontaneous abortion and 180 control couples who did not have a history of spontaneous abortion and had at least one living child. Both parental urinary phenols were measured by ultra-high performance liquid chromatography-tandem mass spectrometry including bisphenol A (BPA), benzophenone-3 (BP-3), 2,3,4-trichlorophenol (2,3,4-TCP), pentachlorophenol (PCP), 4-n-octylphenol (4-n-OP) and 4-n-nonylphenol (4-n-NP). Compared with the low exposure group, there was an increased risk of spontaneous abortion with high paternal urinary PCP concentration [odds ratio (OR) = 2.09, 95% Confidence Interval (CI), 1.05–4.14], and maternal exposure to 4-n-OP and alkylphenol(s) also significantly increased the risk of spontaneous abortion (OR = 2.21, 95% CI, 1.02–4.80; OR = 2.81, 95% CI, 1.39–5.65, respectively). Our study firstly provides the evidence that paternal PCP exposure, maternal 4-n-OP and alkylphenol(s) exposure are associated with spontaneous abortion in humans.     

A study of biochemical parameters among individuals residing in a fluoride endemic area in India

Fluoride (Fl) exerts an inhibitory effect on many metabolic enzymes of various tissues. A study was carried out among individuals residing in a Fl-affected area in India. The biochemical parameters including serum enzyme activities of alkaline phosphatase (ALKP), glutamic pyruvic transaminase (SGPT), lactate dehydrogenase (LDH), and serum creatinine (CRTN) were estimated in 259 subjects compared to 233 controls. The results showed that among the Fl-exposed population, ALKP and SGPT were significantly increased compared to control. The CRTN levels were also significantly increased compared to the control subjects. LDH levels were not markedly altered. Data suggest that Fl exposure affects renal and hepatic functions in endemic areas of India.     

Alterations in some renal parameters of rats induced by aqueous soil extracts

Soil ingestion can be an important route of exposure to contaminants present in the environment. This study examined the effects of exposure to contaminants in aqueous soil extracts from an industrial urban settlement in renal biochemical parameters of treated rats. Male Wistar rats were gavaged with an aqueous soil extract, from the municipality of Rio Grande, Southern Brazil. After exposure, plasma and urine concentrations and plasma protein were assessed compared to rats treated with aqueous soil from relatively unpolluted site (control soil). There was increase in plasma creatinine and total protein in urine, and a decreased glomerular filtration rate in treated rats compared to control. It is possible that Cd, Cr, As, Cu, Pb, Zn, and Ni analyzed in the soil samples and unidentified components may have provoked the observed changes in renal biochemistry of the exposed rat. This may suggest that exposure to contaminated soils can cause damage to the viscera in mammals and it is of public health importance.     

Single-stage versus two-stage models to estimate creatinine corrected urinary analyte concentrations

The ability to detect 'known' differences in urinary analyte concentrations due to gender, age, and race/ethnicity when adjusted for similar differences in urinary creatinine concentrations were evaluated by a single-stage and a two-stage model by ten simulation studies. Log10 transformed values of observed urinary analyte concentration were used as the dependent variable and age, gender, and race/ethnicity were used as the categorical independent variables. In addition, while single-stage model used log10 transformed values of urinary creatinine as a covariate, two-stage model used a correction factor (CF) determined during the first stage of the model by fitting a secondary model for urinary creatinine. Single-stage model was almost always able to statistically significantly detect 'known' differences due to age, gender, and race/ethnicity. On the other hand, two-stage model was able to statistically significantly detect 'known' differences due to age, gender, and race/ethnicity a maximum of 87.2% of the times and as low as 10.6% of the times primarily because of the presence of multicollinearity between CF and urinary creatinine concentrations. Consequently, as long as the sole objective is to estimate the urinary analyte concentrations adjusted for the effect of all factors including urinary creatinine, single-stage models are the models of choice.     

Determination of andrographolide sodium bisulfite in renal microdialysis fluid of rats and toxicokinetic study

Abstract

Andrographolide sodium bisulfite (ASB) widely used for intestinal infections and respiratory tract infections. It has been reported to frequently cause acute renal failure in clinical practice. The purpose of this study was to evaluate the nephrotoxicity and toxicokinetics in Sprague-Dawley rats after a single-dose injection of ASB administered at 100, 600, and 1000?mg/kg via intravenous tail injections. The concentrations in plasma and kidney microdialysates were analyzed by high-performance liquid chromatography. Blood urea nitrogen and creatinine levels in plasma were determined and toxicokinetic parameters were observed. In plasma and kidney, the elimination constant and clearance were decreased and the half-time was increased with increasing dose. Blood urea nitrogen and creatinine were positively correlated with ASB concentration indicating the potential for accumulation in kidney that can eventually lead to damage.     

Fluoride in drinking water exacerbates glomerulonephritis and induces liver damage in ICR-derived glomerulonephritis mice

To evaluate the effects of fluoride on the kidney and the liver of ICR-derived glomerulonephritis (ICGN) mice by using laboratory tests and pathological examinations, fluoride was administered to the ICGN mice at 0, 25, 50, 100, and 150?ppm in drinking water for 4 weeks and to the ICR mice, which have normal kidney function at 0 and 150?ppm. The BUN, creatinine, GOT, and GPT in the serum of each mouse were determined. When a mouse died, the sample from the day closest to the death was assigned for the mean. Pathological changes in the kidney were examined after PAS (periodic acid-Schiff) staining. All of the ICGN mice in the 150?ppm group and one of seven in the 100?ppm group died before the end of week 4, but no ICR mice died. For ICGN mice, the mean value of body weight in the 150?ppm group was significantly lower than those in 0?ppm group and other fluoride-administered groups. The mean values of relative liver and kidney weights in the 100 and 150?ppm groups were significantly lower than those in the control. The mean values of BUN, creatinine, and GPT in the 150?ppm group were significantly higher than those in the control. The thickness of the glomerular capillary wall and the increased mesangial matrix in the kidney were prominent in the fluoride-administered ICGN mice. These results suggested that fluoride severely exacerbated glomerulonephritis and tublar-intestitial changes in ICGN mice.     

Maternal urinary β-core fragment of hCG/creatinine ratios and fetal chromosomal abnormalities in the second trimester of pregnancy

Our aim was to evaluate the potential value of the ratio of the maternal urinary beta-core fragment of human chorionic gonadotropin (βC-hCG) to creatinine (Cr) in discriminating between normal pregnancies and pregnancies associated with fetal chromosomal abnormalities. We hypothesized that pregnancies with fetal chromosomal abnormalities had abnormal quantities of βC-hCG in the urine. The aims of the present study were to investigate retrospectively whether maternal urinary ratios of βC-hCG/Cr are abnormal in women carrying fetuses with chromosome aberrations and to determine normative median values and a reference range for βC-hCG/Cr between 14 and 19 weeks' gestation. Maternal urinary βC-hCG and Cr concentrations were measured in 150 healthy women from 14 to 19 weeks and compared with ten cases of fetal chromosomal abnormalities matched for gestational age. The preliminary cut-off points corresponded to 0·29 multiple of the normal median (MOM) and 2·83 MOM, which were equivalent to the tenth and 90th centiles of the normal range. Of ten cases of fetal chromosomal abnormalities, one out of one (100 per cent) case with trisomy 18 and three of four (75 per cent) cases of variant 9 chromosome had low βC-hCG/Cr (≤0·29 MOM). One of five (20 per cent) cases with Down syndrome had elevated βC-hCG/Cr (≤2·83 MOM). Urinary βC-hCG/Cr ratios obtained in the second trimester may be useful for improved detection efficiency of Down syndrome, trisomy 18, and inversion of chromosome 9. Second-trimester maternal urinary βC-hCG/Cr should be investigated further as a potential marker for fetal chromosome anomalies.