In Vitro Effects of Light on Serum Bilirubin Subfractions Measured by High-Performance Liquid Chromatography: Comparison with Four Routine Methods

1992 ◽  
Vol 38 (10) ◽  
pp. 2124-2129 ◽  
Author(s):  
H Ihara ◽  
H Nakamura ◽  
Y Aoki ◽  
T Aoki ◽  
M Yoshida

Abstract The effects of light on serum bilirubin subfractions in vitro were investigated by HPLC and four routine methods for bilirubin analysis. By HPLC, the rate of photodegradation of unconjugated bilirubin (Bu) was nearly twice that of monoconjugated bilirubin (mBc) and threefold that of diconjugated bilirubin (dBc); delta bilirubin (Bd) was most stable against photoirradiation. In the diazo method, the rate of photodegradation of direct bilirubin was almost the same as that of the sum of mBc, dBc, and Bd determined by the HPLC method. However, the rate of photodegradation of indirect bilirubin was significantly lower (P < 0.001) than that obtained by HPLC, because approximately 30% of the bilirubin photoproducts reacted with the diazo reagent as indirect bilirubin. The rate of photodegradation of total bilirubin determined by the direct spectrometric method was lower than that determined by the diazo method, but equal to that of the total peak areas of HPLC. In the Ektachem method, bilirubin photoproducts affected total bilirubin negligibly, and Bc and Bu positively, so that the value of Bd decreased. In the bilirubin oxidase method, bilirubin photoproducts were oxidized enzymatically by both the total and direct bilirubin reagents. We re-emphasize the importance of shielding serum from light to avoid generating bilirubin photoproducts that interfere with the accurate determination of serum bilirubin subfractions. We also recommend HPLC analysis as a standard method for bilirubin measurement.

Author(s):  
Ishan Joshi ◽  
Amit Sharma

Background: Exchange transfusion (ET) is required in hyperbilirubinemic hospitalized neonates.Methods: The study center at SMS and associated hospitals. The blood bank is licensed and fully equipped. Fall in serum bilirubin minimum of 40 cases was required as the sample size of the present study.Results: The Sex ratio in the neonates undergoing Exchange transfusion was 65% (26) Males and 35% (14) females. The mean values of Hb (g/dl), Hematocrit and Direct bilirubin (mg/dl) of male and female before and after exchange transfusion, were non-significant.Conclusion: Exchange transfusion was required more commonly in male neonates.  The mean values of change in Hb (g/dl), Hematocrit and Direct bilirubin (mg/dl) of males and females before and after exchange transfusion, were non-significant. The mean values of change in Indirect bilirubin (mg/dl) in both males and females before and after exchange transfusion was highly significant.  The mean values of change in Total bilirubin (mg/dl) in males and in females were significant.


Author(s):  
Elvira Dwijayanti ◽  
MI. Diah Pramudianti ◽  
Dian Ariningrum

Congenital hypothyroidism is known to cause prolonged hyperbilirubinemia in neonates. It also correlates with delayedmaturation of the activity of the uridine diphosphate glucoronosyltransferase (UDPG-T) enzyme. Thus, this study wasperformed to analyze the correlation of TSH and FT4 levels with serum bilirubin in neonatal jaundice. This observationalanalytical study with a cross-sectional approach was conducted on 64 neonatal patients with jaundice in Dr. MoewardiGeneral Hospital, Surakarta during September-November 2019. The data comparison and correlation were analyzed withMann-Whitney and the Spearman test. A p-value of < 0.05 was considered significant with 95% Confidence Interval (CI). Thestudy variables comprised of total bilirubin of 12.7 (6.28-23.5) mg/dL, direct bilirubin of 0.8 (0.30-6.61) mg/dL, indirectbilirubin of 11.87 (3.16-22.94) mg/dL, TSH of 4.4 (0.40-23.06) uIU/L, and FT4 of 22.85±7.4 pmol/L. The TSH and FT4 weremoderately correlated with total bilirubin r=-0.444; p=0.001 and r= -0.467; p=0.001), with indirect bilirubin (r= -0.3362;p=0.03 and r=-0.411; p=0.001) and with direct bilirubin (r= -0.257; p=0.040 and r=0.232; p=0.065), respectively. A moderatecorrelation of TSH and FT4 with total and indirect bilirubin, as well as a weak correlation between TSH and direct bilirubinwere found, while no correlation was found between FT4 and with direct bilirubin. Thyroid function screening isrecommended in neonates with jaundice, due to the importance of thyroid hormones in the function and formation oforgans                    


2021 ◽  
Vol 36 (2) ◽  
pp. 172460082110361
Author(s):  
Zhangjun Jia ◽  
Zeyu Zhu ◽  
Ying Wang ◽  
Jing Ding ◽  
Zhenzhong Lin ◽  
...  

Purpose Serum bilirubin plays an important role in antioxidant and anticancer processes. The inverse association between serum bilirubin and cancer risk have been widely reported in multiple cancers. The aim of this retrospective study was to investigate the prognostic impact of serum bilirubin in colorectal cancer patients undergoing surgical resection. Methods The value of serum bilirubin including total bilirubin, direct bilirubin, and indirect bilirubin were tested at pre-operatively in 330 colorectal cancer patients. The optimal cut-off values for these three biomarkers were determined by X-tile program. The relationship between serum bilirubin and outcomes were examined using Kaplan–Meier curves log-rank test, univariate and multivariate cox regression. Moreover, a number of risk factors were used to form a nomogram for evaluating risk of survival. Results The optimal cut-off points of serum total bilirubin, direct bilirubin, and indirect bilirubin were 19.5 μmol/L, 5.0 μmol/L and 8.1 μmol/L, respectively. Elevated total bilirubin and direct bilirubin were significantly associated with overall survival in surgical colorectal cancer patients. Additionally, predictive nomogram including total bilirubin and direct bilirubin for overall survival was established for predicting overall survival in surgical colorectal cancer patients. Conclusions These findings indicated that preoperative elevated total bilirubin and direct bilirubin could be considered as independent prognostic biomarkers for poor overall survival of colorectal cancer patients.


Author(s):  
Armando Raúl Guerra Ruiz ◽  
Javier Crespo ◽  
Rosa Maria López Martínez ◽  
Paula Iruzubieta ◽  
Gregori Casals Mercadal ◽  
...  

Abstract Elevated plasma bilirubin levels are a frequent clinical finding. It can be secondary to alterations in any stage of its metabolism: (a) excess bilirubin production (i.e., pathologic hemolysis); (b) impaired liver uptake, with elevation of indirect bilirubin; (c) impaired conjugation, prompted by a defect in the UDP-glucuronosyltransferase; and (d) bile clearance defect, with elevation of direct bilirubin secondary to defects in clearance proteins, or inability of the bile to reach the small bowel through bile ducts. A liver lesion of any cause reduces hepatocyte cell number and may impair the uptake of indirect bilirubin from plasma and diminish direct bilirubin transport and clearance through the bile ducts. Various analytical methods are currently available for measuring bilirubin and its metabolites in serum, urine and feces. Serum bilirubin is determined by (1) diazo transfer reaction, currently, the gold-standard; (2) high-performance liquid chromatography (HPLC); (3) oxidative, enzymatic, and chemical methods; (4) direct spectrophotometry; and (5) transcutaneous methods. Although bilirubin is a well-established marker of liver function, it does not always identify a lesion in this organ. Therefore, for accurate diagnosis, alterations in bilirubin concentrations should be assessed in relation to patient anamnesis, the degree of the alteration, and the pattern of concurrent biochemical alterations.


Author(s):  
Shohei Kawamoto ◽  
Kosuke Koyano ◽  
Miyo Ozaki ◽  
Takeshi Arai ◽  
Takashi Iwase ◽  
...  

Background Direct-reacting bilirubin concentrations measured using vanadate chemical oxidation method do not exactly match the conjugated bilirubin concentration. One of the causes is the effect of bilirubin photoisomers. However, the quantitative evaluation of the effects of these photoisomers has not been sufficiently conducted. In particular, the influence of bilirubin configurational isomers on direct bilirubin is the most critical factor. Methods Sixteen residual serum samples were used. For quantitative analysis based on the change in direct bilirubin and bilirubin configurational isomer, samples were irradiated via blue light-emitting diodes to suppress the production of bilirubin structural isomers. Total bilirubin and direct bilirubin concentrations were measured using the vanadate chemical oxidation method. Concentrations of 4Z,15Z-bilirubin IXα and its photoisomers were measured using high-performance liquid chromatography. The sum of 4Z,15E-bilirubin IXα and 4E,15Z-bilirubin IXα was notated as bilirubin configurational isomer, and the differences between the measured values of the irradiated and non-irradiated samples were calculated and notated as ΔDB and ΔBCI. Results In non-irradiated and irradiated samples, total bilirubin and direct bilirubin concentrations were 10.73 mg/dL with significant a decrease to 10.60 mg/dL and 0.69 mg/dL with a significant increase to 0.78 mg/dL, while bilirubin configurational isomer values were 1.00 mg/dL and 1.52 mg/dL, respectively. The linear regression equation revealed a significant positive correlation of Y = 0.187X−0.006 between ΔDB (Y) and ΔBCI (X). Conclusion Applying the vanadate chemical oxidation method affected approximately 19% of the bilirubin configurational isomer concentration for direct bilirubin. Extreme caution is necessary when interpreting the measured values of samples indicative of unconjugated hyperbilirubinaemia.


2020 ◽  
Vol 5 (2) ◽  
pp. 98-103
Author(s):  
Asad Ali Khaskheli ◽  
Muhammad Ibrahim Khaskheli ◽  
Allah Jurio Khaskheli ◽  
Arshad Ali Khaskheli

The current study was performed in order to investigate the threat of lactic acidosis in goats worldwide and explore the curative strategies. In this regards a detailed review was performed, however obtained facts were found to be much interesting and valuable. It was indicated by researchers that lactic acidosis is the most common problem in goats throughout the worlds. It represents significant economic loss due to direct and indirect effects. It was further stated that goats with lactic acidosis show decreased body temperature up to 98.1±0.89 °F, rumen and intestinal movement 0.23±0.48/m, rumen pH 4.8±0.07, blood pH 7.1±0.08, increased respiration rate 56.14±7.15/m and heart rate, 136.28±4.71/m. Affected goats also show signs of dyspnea, anorexia, inactivity, incoordination and recumbancy. The glucose level remains 190.14±36.49 mg/dl, total bilirubin 0.75±0.04 mg/dl, direct bilirubin 0.27±0.03mg/dl, indirect bilirubin 0.40±0.03 mg/dl, alanine aminotransferase ALT 36.42±3.04 U/l and alkaline phosphatase 420±3.65 U/l. Furthermore, Glucose level, total bilirubin, direct bilirubin, indirect bilirubin, ALT (SGPT) and alkaline phosphatase increase with treatment of Cassia Fistula, serum biochemical changes rapidly return to normal compared to treatment with Sodium bicarbonate or Magnesium hydroxide. The ruminal juices changes are also significantly improved with the treatment. The changes in the ingesta color, odor and consistency and rumen pH return to normal with the use of Cassia Fistula, Sodium bicarbonate and Magnesium hydroxide. In conclusion, Cassia fistula, Sodium bicarbonate and Magnesium hydroxide could be used as valuable strategies against lactic acidosis in goats. These therapies have been proved to be effective for treating the acidosis in goats. 


1983 ◽  
Vol 29 (3) ◽  
pp. 466-469 ◽  
Author(s):  
D M Nathan ◽  
T B Francis ◽  
J L Palmer

Abstract We investigated the in vivo and in vitro effects of aspirin on several clinical assays of glycosylated hemoglobin. Acetylation of hemoglobin falsely increased the glycosylated hemoglobin fraction measured by "high-performance" liquid chromatography and electrophoresis, but isoelectric focusing and colorimetric techniques differentiated between acetylated and glycosylated fractions. Aspirin ingestion may result in an apparent increase in glycosylated hemoglobin measured with common clinical assays.


2020 ◽  
Vol 9 (12) ◽  
Author(s):  
Jiancheng Wang ◽  
Xianglin Zhang ◽  
Zhuxian Zhang ◽  
Yuanyuan Zhang ◽  
Jingping Zhang ◽  
...  

Background Data on the association between serum bilirubin and the risk of stroke are limited and inconclusive. We aimed to evaluate the association between serum bilirubin and the risk of first stroke and to examine any possible effect modifiers in hypertensive patients. Methods and Results Our study was a post hoc analysis of the CSPPT (China Stroke Primary Prevention Trial). A total of 19 906 hypertensive patients were included in the final analysis. Cox proportional hazards models were used to estimate the hazard ratios (HRs) and 95% CIs for the risk of first stroke associated with serum bilirubin levels. The median follow‐up period was 4.5 years. When serum total bilirubin was assessed as tertiles, the adjusted HR of first ischemic stroke for participants in tertile 3 (12.9–34.1 μmol/L) was 0.75 (95% CI, 0.59–0.96), compared with participants in tertile 1 (<9.3 μmol/L). When direct bilirubin was assessed as tertiles, a significantly lower risk of first ischemic stroke was also found in participants in tertile 3 (2.5–24.8 μmol/L) (adjusted HR, 0.77; 95% CI, 0.60–0.98), compared with those in tertile 1 (<1.6 μmol/L). However, there was no significant association between serum total bilirubin (tertile 3 versus 1: adjusted HR, 1.45; 95% CI, 0.89–2.35) or direct bilirubin (tertile 3 versus 1: adjusted HR, 1.27; 95% CI, 0.76–2.11) and first hemorrhagic stroke. Conclusions In this sample of Chinese hypertensive patients, there was a significant inverse association between serum total bilirubin or direct bilirubin and the risk of first ischemic stroke.


Angiology ◽  
2016 ◽  
Vol 68 (5) ◽  
pp. 428-432 ◽  
Author(s):  
Ertan Vuruşkan ◽  
Erhan Saraçoğlu ◽  
İrfan Veysel Düzen

The relationship between bilirubin levels and peripheral artery disease has been documented. Our aim was to demonstrate the possible relationship between serum bilirubin levels and abdominal aortic aneurysms (AAAs). The study included 219 patients, 110 had a previous diagnosis of AAA and 109 patients were normal controls. Only patients with AAAs which had a size of 40 to 54 mm were included in the study. Baseline laboratory values and 2 computerized tomographic measurements 12 months apart were recorded. Patients with AAA had significantly higher white blood cell (WBC) counts and neutrophil–lymphocyte ratio (NLR) but lower total and direct bilirubin levels compared with the control patients ( P < .05). Multivariate logistic regression analysis showed that WBC, NLR, and total and direct bilirubin levels were independent predictors of the presence of an AAA ( P = .03, P = .001, P = .001, and P = .001, respectively). White blood cells and total bilirubin level were independent predictors of a rapidly enlarging AAA (>10 mm/y, P = .002 and P < .001, respectively). This study demonstrated that increased WBC and decreased total bilirubin levels were independent predictors of an AAA, especially the subgroup in which the AAA was rapidly expanding.


2011 ◽  
Vol 399-401 ◽  
pp. 1067-1070
Author(s):  
Chun Yan Li ◽  
Cong Cong Hu ◽  
Zhi Guo Wen ◽  
Sheng Xiong Dong

The method of high performance liquid chromatography (HPLC) is established to determine the content of antibacterial agent — ciprofloxacin (CF) in the degradation solution of ciprofloxacin-polyurethane (CFPU) and investigate the in vitro degradation kinetics by plotting and fitting the cumulative release curves to inspect the effects of different medium and different concentrations on drug release. The results showed that the HPLC method is accurate, reliable and simple. The drug-release of CFPU was bioresponsive and could be accorded with first order kinetics. It was observed that CF was released from CFPU by a combination of diffusion and erosion mechanism, mainly in the manner of diffusion in the absence of infection while erosion mechanism in the presence of infection.


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