Automation of Harboe method for the measurement of plasma free hemoglobin

Chung, Hee-Jung; Chung, Jae-Woo; Yi, Joowon; Hur, Mina; Lee, Tae Hwan; Hwang, Sang‐Hyun; Song, Yoon Kyung; Lee, Dong Hoon

doi:10.1002/jcla.23242

Cited by 8 publications

(7 citation statements)

References 16 publications

(20 reference statements)

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“…RBCs were then pelleted by centrifugation at 200 g for 2 min. RBC pellets were lysed in 0.2% Triton-X-100 and both supernatants and pellet lysates were assessed for Hb content by spectrophotometry (Packard SpectraCount Absorbance Microplate Reader) at 450 nm as oxy and deoxy forms of hemoglobin show main peaks of light absorption around 450 nm ( Ratanasopa et al, 2015 ; Chung et al, 2020 ). For each medium, Hb in the supernatant was then expressed as ratio of the sum of Hb in the supernatant and the pellet.…”

Section: Methodsmentioning

confidence: 99%

Interplay Between Plasma Membrane Lipid Alteration, Oxidative Stress and Calcium-Based Mechanism for Extracellular Vesicle Biogenesis From Erythrocytes During Blood Storage

et al. 2020

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The shedding of extracellular vesicles (EVs) from the red blood cell (RBC) surface is observed during senescence in vivo and RBC storage in vitro . Two main models for EV shedding, respectively based on calcium rise and oxidative stress, have been proposed in the literature but the role of the plasma membrane lipid composition and properties is not understood. Using blood in K + /EDTA tubes stored for up to 4 weeks at 4°C as a relevant RBC vesiculation model, we showed here that the RBC plasma membrane lipid composition, organization in domains and biophysical properties were progressively modified during storage and contributed to the RBC vesiculation. First, the membrane content in cholesterol and linoleic acid decreased whereas lipid peroxidation and spectrin:membrane occupancy increased, all compatible with higher membrane rigidity. Second, phosphatidylserine surface exposure showed a first rapid rise due to membrane cholesterol decrease, followed by a second calcium-dependent increase. Third, lipid domains mainly enriched in GM1 or sphingomyelin strongly increased from the 1st week while those mainly enriched in cholesterol or ceramide decreased during the 1st and 4th week, respectively. Fourth, the plasmatic acid sphingomyelinase activity considerably increased upon storage following the sphingomyelin-enriched domain rise and potentially inducing the loss of ceramide-enriched domains. Fifth, in support of the shedding of cholesterol- and ceramide-enriched domains from the RBC surface, the number of cholesterol-enriched domains lost and the abundance of EVs released during the 1st week perfectly matched. Moreover, RBC-derived EVs were enriched in ceramide at the 4th week but depleted in sphingomyelin. Then, using K + /EDTA tubes supplemented with glucose to longer preserve the ATP content, we better defined the sequence of events. Altogether, we showed that EV shedding from lipid domains only represents part of the global vesiculation mechanistics, for which we propose four successive events (cholesterol domain decrease, oxidative stress, sphingomyelin/sphingomyelinase/ceramide/calcium alteration and phosphatidylserine exposure).

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Section: Methodsmentioning

confidence: 99%

Interplay Between Plasma Membrane Lipid Alteration, Oxidative Stress and Calcium-Based Mechanism for Extracellular Vesicle Biogenesis From Erythrocytes During Blood Storage

et al. 2020

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“…Normal range of Hb for men is 13.5-17.5 grams per deciliter and for women it is 12.0-15.5 grams per deciliter (Ajibola, 2012). depending on your age and gender a normal hemoglobin level is 11 to 18 grams per deciliter (g / dl) (Chung et al, 2020). Yet a healthy range is from 7 to 8 g / dl.…”

Section: Resultsmentioning

confidence: 99%

“…Hemoglobin measurement is now performed in several laboratories using Automatic Hematology Analyzers but also in many other laboratories using Sahli 's method a.k.a(organisation mondiale de la santé et al, 2003; Nayak, Rai and gupta, 2011). Acid Hematin method, Cyanmethemoglobin method (Cmg) a.k.a drabkin 's method widely used to evaluate the concentration of hemoglobin in the blood of patients (Chung et al, 2020). The theory of the Sahli method or acid hematin method is very simple that the hemoglobin present in RBCs is converted into acid hematin, which is a dark brown colored substance, when the blood is added to N/10 Hydrochloric acid ( HCl)(kawthalkar, 2012).The color of the shaped acid hematin complex corresponds to the concentration of hemoglobin in the blood and is matched to the norm which is a reference brown glass supplied in the Sahli 's apparatus by diluting with N/10 hydrochloric acid or distilled water until the acid hematin complex color matches the norm colour (Sood, 1985).…”

Section: Methodsmentioning

confidence: 99%

Comparison of Hb and PCV Values in Manual Methods – A Prospective Study

Ananya¹

2020

Biosci. Biotech. Res. Comm

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Hemoglobin iron-containing metalloprotein expressed in red for all vertebrate blood cells (erythrocytes), as well as some invertebrate tissues. A healthy person has 12 to 20 grams of hemoglobin in every 100 milliliters of blood. Hematocrit is a blood test measuring the percentage concentration of red blood cells (RBC) in the blood. The measurement will depend on the number and size of cells in red bloods. It is part of the complete results of a person's blood count, along with hemoglobin concentration, white blood cell count, and platelet count. The volume of packed cells (PCV) is a measure of the proportion of blood formed by cells. The value is expressed as one percentage or fraction of blood cells. For example, a 40 per cent PCV means that in 100 milliliters of blood there are 40 milliliters of cells. This study is to compare the Hb and PCV values derived from direct manual and automated methods. Nowadays PCV values are estimated through automated methods. If there are any electrical supply problems, low budget labs, unavailability of the wintrobe's tube, in a practical college set up we have sahli's haemoglobinometer , with this we can be able to calculate the PCV valve. But our results show that the automated PCV values are higher than the manual calculated PCV values and the automated methods are more accurate to follow. Also we found that the manual calculated PCV method is not a better alternative for Automated PCV method.

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“…The spectrophotometric method of measuring PFHb has a precision of SD 5.2 mg/dL and CV of 9.4% for PFHb value of 5.2 mg/dL in the range of (0.3-62.5) mg/dL. This method cannot be automated because the technician has to estimate the PFHb by measuring the absorbance at three wavelengths (380 nm, 415 nm, 450 nm) 31 . The Hb measuring device ADVIA 120 Hematology analyzer (Siemens AG, Germany) has an accuracy of SD 0.14 g/dL and CV 0.93% for Hb value of 15 g/dL in the range of (0.0-22.5) g/dL.…”

Section: Discussionmentioning

confidence: 99%

Point-of-care testing of plasma free hemoglobin and hematocrit for mechanical circulatory support

Shin

Lee

Shin

et al. 2021

Sci Rep

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Hematological analysis is essential for patients who are supported by a mechanical circulatory support (MCS). The laboratory methods used to analyze blood components are conventional and accurate, but they require a mandatory turn-around-time for laboratory results, and because of toxic substances, can also be hazardous to analysis workers. Here, a simple and rapid point-of-care device is developed for the measurement of plasma free hemoglobin (PFHb) and hematocrit (Hct), based on colorimetry. The device consists of camera module, minimized centrifuge system, and the custom software that includes the motor control algorithm for the centrifuge system, and the image processing algorithm for measuring the color components of blood from the images. We show that our device measured PFHb with a detection limit of 0.75 mg/dL in the range of (0–100) mg/dL, and Hct with a detection limit of 2.14% in the range of (20–50)%. Our device had a high correlation with the measurement method generally used in clinical laboratories (PFHb R = 0.999, Hct R = 0.739), and the quantitative analysis resulted in precision of 1.44 mg/dL for PFHb value of 14.5 mg/dL, 1.36 mg/dL for PFHb value of 53 mg/dL, and 1.24% for Hct 30%. Also, the device can be measured without any pre-processing when compared to the clinical laboratory method, so results can be obtained within 5 min (about an 1 h for the clinical laboratory method). Therefore, we conclude that the device can be used for point-of-care measurement of PFHb and Hct for MCS.

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Automation of Harboe method for the measurement of plasma free hemoglobin

Cited by 8 publications

References 16 publications

Interplay Between Plasma Membrane Lipid Alteration, Oxidative Stress and Calcium-Based Mechanism for Extracellular Vesicle Biogenesis From Erythrocytes During Blood Storage

Interplay Between Plasma Membrane Lipid Alteration, Oxidative Stress and Calcium-Based Mechanism for Extracellular Vesicle Biogenesis From Erythrocytes During Blood Storage

Comparison of Hb and PCV Values in Manual Methods – A Prospective Study

Point-of-care testing of plasma free hemoglobin and hematocrit for mechanical circulatory support

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