hemoglobin isosbestic point

New smart spectrophotometric methods have been recently evolved with no limitation. The first derivative of the above ratio spectra was then recorded using scaling factor=1 and =8 and a calibration graph between the amplitude at 219nm versus the corresponding concentrations of CAF was constructed. -. Detectability of low-oxygenated regions in human muscle tissue using near-infrared spectroscopy and phantom models. Ratio difference-isosbestic points (RD-ISO) is considered as an extension to ratio difference method [17]. Mohamed, E.H., Lotfy, H.M., Hegazy, M.A. Epub 2017 Jun 22. The three isosbestic wavelength ratio pairs that best correlated to total hemoglobin concentration independent of saturation and scattering were 545/390, 452/390, and 529/390 nm. Principles of pulse oximetry. doi: 10.1093/tropej/fmaa111. Pharmacol Rev 51:8388, CAS While for PAP the correlation was obtained between the amplitude difference at 311 and 318nm at which PCT have the same amplitude \(\left( {\Delta {\text{P}}_{\text{PCT}} = {\text{P}}_{ 1} - {\text{P}}_{ 2} = {\text{zero}}} \right)\) and the corresponding PAP concentration was plotted from which its concentration could be determined as shown in Fig. 138, 99103 (2015).CNNSBV10.1016/j.clineuro.2015.07.021 The oxygen saturation is estimated by measuring the transmission of light through the pulsatile tissue bed. While literature survey reveals that no methods have been reported for the simultaneous determination of the four components under study. Principles and Problems in Physical Chemistry for Biochemists. Spectrophotometry & Spectrofluorimetry. Results: The methods were validated according to the ICH guidelines, accuracy, precision, repeatability, were found to be within the acceptable limits. sharing sensitive information, make sure youre on a federal 43 Because our device was operating near the isosbestic point of hemoglobin, [44][45] [46] we . As a consequence, all mixtures of A and B also show the same absorption at that frequency, and all the spectra along, e.g., a pH titration from A to B, plotted one on top of the other, cross at the isosbestic point. Materials (Basel). a Ratio spectra of DRO (solid line), CAF (dashed line), PCT (dotted line) and their ternary mixture (dashed dotted line) containing 10g/mL of each using normalized CAF spectrum as a divisor. Analysis of complex mixture containing three or more components represented a challenge for analysts. Caffeine (CAF) 1,3,7-Trimethylpurine-2,6-Dione, is an adenosine receptor antagonist and adenosine 3,5cyclic monophosphate (cAMP) phosphodiesterase inhibitor, thus levels of cAMP increase in cells following treatment with caffeine [2, 3]. Spectrochim Acta A Mol Biomol Spectrosc 140:600613, Lotfy HM, Hegazy MA, Mowaka S, Mohamed EH (2015) Novel spectrophotometric methods for simultaneous determination of amlodipine, valsartan and hydrochlorothiazide in their ternary mixture. The isosbestic point of the HB/HbO2 absorption spectrum is circled in white . A calibration graph was constructed relating the absorbance of zero order spectra (D0) of DRO at 228.5nm versus the corresponding concentrations. The proposed spectrophotometric methods are also considered to be fast and time saving where the analysis of the quaternary or the ternary mixture takes few seconds once calibration graphs were constructed and regression equations are computed where all the reported chromatographic techniques needs at least 10min in a single run to resolve the ternary mixture. Absorption spectra of Hb, HbO2, H2O and other chromophores in NIR range (redrawn after Murkin and Arango 2009 [11]). By scanning the absorption spectra of DRO, CAF, PCT and PAP in the solution of dosage forms in methanol, severely overlapped spectral bands were observed in the wavelength region of 200300nm; which hindered their direct determination (Fig. By substituting the absorbance difference A (AisoA2) between the two selected wavelengths of the mixture spectrum in the above equation, the absorbance Apostulated corresponding to the absorbance of CAF only at Aiso was obtained. However, due to the low satisfaction rate, invasive solutions are still assumed as the gold standard. Kahraman S, Kayali H, Atabey C, Acar F, Gocmen S. J Trauma. National Library of Medicine By John W. Moore, Learn how and when to remove this template message, A/D variant of the total syntheses of vitamin B, https://en.wikipedia.org/w/index.php?title=Isosbestic_point&oldid=1137892749, This page was last edited on 6 February 2023, at 23:40. The quantity of PAP must be strictly controlled as it is reported to have nephrotoxic and teratogenic effects [7]. Progressive evaluation in spectroscopic sensors for non-invasive blood haemoglobin analysis-a review. The (D0) spectra of CAF were divided by the normalized spectrum of PCT, and then two calibration graphs were constructed. 11b. Note that distance from phantom surface through CSF layer is 1.1cm. This study aims at an optimistic optical spectroscopic technique to measure hemoglobin concentration and conditional usability of non-invasive blood parameters' diagnostics at point-ofcare. saturation). Isosbestic points are used in medicine in a laboratory technique called oximetry to determine hemoglobin concentration, regardless of its saturation. Spectrochim Acta A Mol Biomol Spectrosc 148:328337, Lamie NT, Yehia AM (2015) Development of normalized spectra manipulating spectrophotometric methods for simultaneous determination of dimenhydrinate and cinnarizine binary mixture. Figure 4: An example of isosbestic point (CC BY-4.0; Heesung . The structures of the studied drugs are presented in Fig. As shown in (Fig. 12a. The concentrations of CAF and PCT were calculated using the corresponding unified regression equation (obtained by plotting the absorbance of the zero order spectra of CAF or PCT at iso 263.6nm against the corresponding concentrations). In spectroscopy, an isosbestic point is a specific wavelength, wavenumber or frequency at which the total absorbance of a sample does not change during a chemical reaction or a physical change of the sample. Serum ferritin testing is . The non-constant absorption (AC) is the result of pulsatile blood pulsations. The proposed methods also showed the significance of isoabsorptive point, normalized spectra as divisors and dual wavelengths as powerful tools that could either be used alone or in combination with each other for the resolution of severely overlapped spectra without preliminary separation. The validity of the proposed methods was further assessed by applying the standard addition technique as presented in Table3. A pair of substances can have several isosbestic points in their spectra. Oxyhaemoglobin and deoxyhaemoglobin have (not exclusively) isosbestic points at 586 nm and near 808 nm. A study of different novel and smart spectrophotometric techniques for resolution of severely overlapping spectra were presented in this work utilizing isosbestic points present in different absorption spectra, normalized spectra as a divisor and dual wavelengths. Spectrochim Acta A Mol Biomol Spectrosc 136:17861796, Din E (1999) The spectrophotometric multicomponent analysis of a ternary mixture of ascorbic acid, acetylsalicylic acid and paracetamol by the double divisor-ratio spectra derivative and ratio spectra-zero crossing methods. The concentration of each drug was calculated using its specified methods. Two distinguishable light emitting sources (810 nm and 1300 nm) are utilized at isosbestic points with a single photodetector (800-1700 nm). The adopted techniques adopted of successive and progressive steps manipulating zero /or ratio /or derivative spectra. a Ratio spectra of 4 binary mixtures of DRO and CAF in different concentrations using the sum of normalized spectra of DRO and CAF as double divisor showing the obtained constant regions. Moreover, principal component analytical model with multivariate linear regression offers mean bias of 0.036 and -0.316 g/dL, precision of 0.878 and 0.838 and limits of agreement from -1.685 to 1.758 g/dL and -1.790 to 1.474 g/dL for 18 and 21 principal components, respectively. Isosbestic points are also used in clinical chemistry, as a quality assurance method, to verify the accuracy in the wavelength of a spectrophotometer. Zero order absorption spectra of 10g/mL PCT (dotted line) and CAF (dashed line) showing 3 isoabsorptive points at 226.9 263.6 and 292nm and the binary mixture of CAF and PCT 10g/mL of each, a Ratio absorption spectra of 10g/mL PCT (dotted line), 10g/mL CAF (dashed line) and the binary mixture of CAF and PCT 5g/mL of each (dotted straight line) obtained after division by the normalized spectra of PCT. A quaternary mixture of drotaverine (DRO), caffeine (CAF), paracetamol (PCT) and para-aminophenol (PAP) was taken as an example for application of the proposed techniques without any separation steps. Lastly, dithionite has been shown to increase the 800 nm isosbestic point between oxyHb and deoxyHb due to side reactions . Chemistry Central Journal 11, 43 (2017). Traditional invasive techniques have . For this PubMedGoogle Scholar. To cancel the contribution of PCT at both selected wavelengths, induced dual wave length method was adopted by calculating an equality factor for pure PCT at two selected wave lengths of CAF \(\left( {{\text{F}} \; = \; \left[ {{\text{P}}_{ 2 6 5} /{\text{P}}_{ 2 9 5} } \right] \; = \; 5. The DiaSpect Hemoglobin T and DiaSpect Tm use a slightly different technology. 10b. Bethesda, MD 20894, Web Policies Merck and Co Inc, Whitehouse Station, Sweetman SC (2009) Martindale: the complete drug reference, 33rd edn. Note that in (a), some part of the hematoma is directly below one of the fibers as long as the distance from the center is. Spectrophotometry at that wavelength measures total concentration of hemoglobin, . We also acknowledge previous National Science Foundation support under grant numbers 1246120, 1525057, and 1413739. official website and that any information you provide is encrypted Different applications of isosbestic points, normalized spectra and dual wavelength as powerful tools for resolution of multicomponent mixtures with severely overlapping spectra. In order to determine of PCT, the same procedures were applied as described for CAF. {Site map} {Site disclaimer} {Privacy Policy} {Terms and conditions}, Add this article to my examination home page. . eCollection 2020. Each drug in the quaternary mixture can be determined and analysed by more than one method using different approaches. Subtracting the postulated absorbance of CAF at Aiso from the practically recorded absorbance [ARecorded] at Aiso to get that corresponding to PCT. 2017;2017:1846830. doi: 10.1155/2017/1846830. This site needs JavaScript to work properly. The proposed procedures are accurate, simple and reproducible and yet economic. 3, but it was difficult to accurately measure the amplitude of DRO at its extended region due to its low absorptivity, so derivative transformation was adopted to overcome this problem. A low hemoglobin count can also be due to blood loss, which can occur because of: Bleeding in your digestive tract, such as from ulcers, cancers or hemorrhoids; Frequent blood donation; Menorrhagia (heavy menstrual bleeding) (heavy menstrual bleeding-although even normal menstrual bleeding may cause a slightly low hemoglobin count) First order absorption spectra of 10g/mL DRO (solid line), 10g/mL PCT (dotted line), 10g/mL CAF (dashed line) and 10g/mL PAP (dashed dotted line), Zero order absorption spectra of DRO in mixtures (2, 6, 10, 12, and 20g/mL). 1. Talanta 37:347351, International conference on harmonization (ICH), Q2B: validation of analytical procedures: methodology, vol 62. Dark-shaded regions in (a) and (c)represent blood, light-shaded region in (b)represents bloodCSF mixture; hashed regions represent CSF layer. Different applications of isosbestic points, normalized spectra and dual wavelength as powerful tools for resolution of multicomponent mixtures with severely overlapping spectra, $${\text{P}}_{ 1} = \left[ {{\text{C}}_{\text{x}} } \right]\; + \;\left[ {{\text{C}}_{\text{Y}} } \right]\; + \;\left[ {{\text{a}}_{\text{z1}} {\text{C}}_{\text{z}} } \right]/{\text{a}}_{\text{x}}$$, $${\text{P}}_{ 2} = \;\left[ {{\text{C}}_{\text{x}} } \right]\; + \;\left[ {{\text{C}}_{\text{Y}} } \right]\; + \;\left[ {{\text{a}}_{\text{z2}} {\text{ C}}_{\text{z}} } \right]/{\text{a}}_{\text{x}}$$, $${\text{P}}_{ 1} - {\text{P}}_{ 2} = \left( {\frac{{a_{z} C_{z} }}{{a_{x} }}} \right)1 - \left( {\frac{{a_{z} C_{z} }}{{a_{x} }}} \right)2$$, $${\text{Pm}}_{ 1} = {\text{P}}_{\text{X1}} \; + \;{\text{P}}_{\text{Y1}} \quad {\text{at }}\lambda_{ 1}$$, $${\text{Pm}}_{ 2} = {\text{P}}_{\text{X2}} \; + \;{\text{P}}_{\text{Y2}} \quad {\text{at }}\lambda_{ 2}$$, $${\text{F}}_{\text{Y}} = {\text{P}}_{\text{Y1}} /{\text{P}}_{\text{Y2}}$$, $$\therefore {\text{P}}_{\text{Y1}} = {\text{F}}_{\text{Y}} {\text{P}}_{\text{Y2}}$$, $${\text{Pm}}_{ 1} = {\text{P}}_{\text{X1}} \; + \;{\text{F}}_{\text{Y}} {\text{P}}_{\text{Y2}}$$, $${\text{F}}_{\text{Y}} {\text{Pm}}_{ 2} = {\text{F}}_{\text{Y}} {\text{P}}_{\text{X2}} \; + \;{\text{F}}_{\text{Y}} {\text{P}}_{\text{Y2}}$$, $$\Delta {\text{P }}\left( {{\text{Pm}}_{ 1} \;{-}\;{\text{F}}_{\text{Y}} {\text{Pm}}_{ 2} } \right) = {\text{A}}_{\text{X1}} \; - \;{\text{F}}_{\text{Y}} {\text{A}}_{\text{X2}}$$, $${\text{Pm}} = \frac{{a_{X} C_{X} }}{{a_{Z} + a_{W} }}\; + \,\frac{{a_{Y} C_{Y} }}{{a_{Z} + a_{W} }}\; + \;{\text{constant}}$$, $${\text{P}}_{ 1} = \frac{{a_{X} C_{X} }}{{[a_{Z} + a_{W} ]1}} + \frac{{a_{Y} C_{Y} }}{{[a_{Z} + a_{W} ]1}} + {\text{constant}}$$, $${\text{P}}_{ 2} = \frac{{a_{X} C_{X} }}{{[a_{Z} + a_{W} ]2}} + \frac{{a_{Y} C_{Y} }}{{[a_{Z} + a_{W} ]2}} + {\text{constant}}$$, $$\frac{{a_{Y} C_{Y} }}{{[a_{Z}\,+\,a_{W} ]1}} = \frac{{a_{Y} C_{Y} }}{{[a_{Z}\,+\,a_{W} ]2}}$$, $${\text{P}}_{ 1} - {\text{P}}_{ 2} = \left( {\frac{{a_{X} C_{X} }}{{a_{Z + } a_{W} }}} \right)1 - \left( {\frac{{a_{X} C_{X} }}{{a_{Z + } a_{W} }}} \right)2$$, \(\left( {{\text{F}} \; = \; \left[ {{\text{P}}_{ 2 6 5} /{\text{P}}_{ 2 9 5} } \right] \; = \; 5. Google Scholar, Hegazy MA, Eissa MS, Abd El-Sattar OI, Abd El-Kawy MM (2014) Determination of a novel ACE inhibitor in the presence of alkaline and oxidative degradation products using smart spectrophotometric and chemometric methods. , Clinical evaluation of a portable near-infrared device for detection of traumatic intracranial hematomas, J. Neurotrauma 27(9), 15971604 (2010).JNEUE410.1089/neu.2010.1340 The prepared solutions were found to be stable without any degradation when stored in the dark in the refrigerator at 4C for 1week except for PAP which should be freshly prepared. Neurol. Effect of volume/thickness and depth on detectability in intracerebral hematoma phantoms. A correlation was obtained between the amplitude difference at 315 and 336nm at which PCT have the same amplitude \(\left( {\Delta {\text{P}}_{\text{PCT}} = {\text{P}}_{ 1} - {\text{P}}_{ 2} = {\text{zero}}} \right)\) and the corresponding DRO concentration was plotted from which its concentration could be determined as shown in Fig. Hemoglobin; anemia; isosbestic point; non-invasive sensing; optical systems; sliding mechanism. Finally another novel method for simultaneous determination of quaternary mixtures was proposed and named double divisor-ratio difference-dual wave length (DD-RD-DW). For determination of PCT, the difference between the amplitude of the above ratio spectra obtained after dividing the spectrum of the ternary mixture by the normalized spectrum of CAF at the two selected isosbestic points (219.2 and 252nm) between CAF and DRO was corresponding to PCT concentration only as shown in Fig. If one does nothing to correct for haemoglobin concentration, the pulse oximeter will become confused and measure something spurious, particularly as the patient becomes more hypoxic. Biomed Opt Express. Table4 showed statistical comparisons of the results obtained by the proposed methods and reported method for DRO [23], and official methods for CAF [24] and PCT [25]. The diodes flash at approximately 30 times per second. Measurements of these phantoms are made with a commercial NIRS-based hematoma detector to quantify the effect of hematoma type, depth, and size, as well as measurement repeatability and detector positioning relative to the hematoma. The volumes were completed with methanol. The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). The concentration of DRO was determined using amplitude modulation method from the straight line parallel to the x-axis in the extended region at 310400nm for DRO as shown in Fig. The photo detector generates a voltage proportional to the transmitted light. 10b. Sarkar PK, Pal S, Polley N, Aich R, Adhikari A, Halder A, Chakrabarti S, Chakrabarti P, Pal SK. The calculated t and F values were less than the theoretical ones indicating that there was no significant difference between them with respect to accuracy and precision. The coordinates for an isosbestic point is given (470 nm, =47.7 10 2 m 2 mol 1). PCT was determined using six different methods; advanced absorbance subtraction, advanced amplitude modulation, simultaneous derivative ratio, ratio difference-isosbestic points, induced ratio difference and double divisor-ratio difference-dual wavelength. Federal government websites often end in .gov or .mil. Bethesda, MD 20894, Web Policies Talanta 48:11451157, Article 9a. The diodes are switched on in sequence, with a pause . At the isosbestic point the total absorbance of a solution of the two ions is independent of their . Two light-emitting diodes produce beams at red and infrared frequencies (660 nm and 940 nm, respectively). The ratio spectra obtained using the normalized spectrum of the more extended component Z as a divisor generated a constant value of its concentration along the whole spectra that can be measured from the extended region parallel to the X axis. This is done by measuring the spectra of a standard substance at two different pH conditions (above and below the pKa of the substance). Photographs of phantom layers incorporating (a)5- and (b)8.5-cm-diameter hematoma-simulating inclusions, as well as the (c)hematoma detector on its scaffold and (d)during a handheld measurement. The spectra of the prepared standard solutions were scanned from 200 to 400nm and stored in the computer against methanol as a blank. The red dashed line indicates the detection threshold. eCollection 2021. Disclaimer. 2006 Dec;61(6):1480-3. doi: 10.1097/01.ta.0000197616.10279.48. The requirement for an isosbestic point to occur is that the two species involved are related linearly by stoichiometry, such that the absorbance is invariant at a certain wavelength. Stock standard solutions(1mg/mL) stock solution of each of DRO, CAF, PCT and PAP in methanol were prepared. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. PLoS One. Careers. et al. The AGA defines anemia as a hemoglobin level of less than 13 g per dL (130 g per L) in men and less than 12 g per dL (120 g per L) in patients who are not pregnant. Schematic of phantoms representing (a) epi/subdural, (b) subarachnoid, and (c) intracerebral hematomas. The appearance of an isosbestic point in a reaction demonstrates that an intermediate is NOT required to form a product from a reactant. The proposed spectrophotometric methods were validated in compliance with the ICH guidelines [22], as shown in Table1. The AAM method was applied by dividing the spectrum of the binary mixture by the normalized divisor of PCT to obtain the ratio spectra (Fig. Does the absorbance at the isosbestic point depend on indicator concentration or PH? -. eCollection 2020 Jun 1. Intracerebral hematomas are detectable to a maximum depth of 2.5 cm, depending on thickness and diameter. All authors read and approved the final manuscript. The stored (D0) spectra of DRO were divided by (a) the normalized spectrum of CAF, (b) the normalized spectrum of DRO, (c) sum of normalized spectrum of CAF and PAP, separately. Similarly, derivative transformation technique was adopted to obtain the D0 of PAP by dividing the spectrum of the above resolved ternary mixture by the first derivative of normalized spectrum of PAP (d/d) [aPAP], and then the constant generated in the region 310330nm was multiplied by the normalized spectrum of PAP [aPAP]. 2023 BioMed Central Ltd unless otherwise stated. The standards used include potassium dichromate (isosbestic points at 339 and 445 nm), bromothymol blue (325 and 498 nm) and congo red (541 nm). A remarkable amplitude difference between the two selected wavelengths in the ratio spectra of pure X should be present. The same procedures were applied for determination of CAF where the absorption spectrum of the mixture was divided by the absorption spectrum of the normalized spectra of PCT as divisor and the difference between the amplitude at the two selected isosbestic points (233.8 and 273.7nm) between DRO and PCT was corresponding to CAF concentration only as shown in Fig. The modified Beer-Lambert law was applied for calculating changes in absorption coefficient ( a ). The https:// ensures that you are connecting to the This site needs JavaScript to work properly. The derivative transformation was applied to obtain the (D0) of DRO by dividing the spectrum of the quaternary mixture by the first derivative of normalized spectrum of DRO (d/d) [aDRO], and then the constant generated in the region 360380nm was multiplied by the normalized spectrum of DRO [aDRO] where the absorbance of DRO can be measured at its 228.5nm (max) giving maximum sensitivity and minimum error as shown in Fig. Paracetamol (PCT) N-(4-hydroxyphenyl) acetamide, also known as acetaminophen PAR is widely used as analgesic and antipyretic for the relief of fever, headaches and minor pains. Suppose the amplitudes at the two selected wavelength are P1 and P2 at 1 and 2 (where Y has the same amplitude), respectively, then; The concentration of X is calculated using the regression equation representing the linear correlation between the differences of ratio spectra amplitude at the two selected wavelengths to the corresponding concentrations of drug (X). We developed a ratiometric method capable of estimating total hemoglobin concentration from optically measured diffuse reflectance spectra. The properties of isosbestic points are considered with special reference to systems containing more than two absorbing species and the problems of diagnosing the positive existence of an isosbestic point when a set of spectra cross, or appear to cross, at a point. Calibration graphs were constructed by plotting (a) the difference between the amplitudes at [263.6 and 291.8nm], (b) the constant values measured from 310400nm, (c) the difference between the amplitudes at [315 and 336nm] versus the corresponding DRO concentrations, respectively. These reports proposed spectrophotometric [8, 9], TLC [9] and high performance liquid chromatography (HPLC) [8, 10, 11]. isosbestic: ( 'sos-bes'tik ), Denoting the wavelength of light at which two related compounds have identical extinction coefficients; for example, the wavelength at which the absorption spectra of hemoglobin and oxyhemoglobin cross is their isosbestic point. \(\epsilon\) is the molar extinction coefficient or molar absorptivity (or absorption coefficient). Bullock M. R., et al. Three different novel, simple and accurate methods were adopted for simultaneous determination of CAF and PCT in presence of each other either in bulk, in different dosage forms as binary mixture and in presence of other components after their resolutions. Shukla D, Konar S, Devi BI, Padmasri G, Jayanna R, Suresh M, Lakshita B. Clipboard, Search History, and several other advanced features are temporarily unavailable. The advantage of this method over the advanced absorbance subtraction method is the complete cancelling of the interfering component in the form of constant where the difference at any two points along its ratio spectrum will be equal to zero. FOIA To cancel the contribution of Y at the two selected wavelengths upon obtaining the ratio difference, the equality factor of pure ratio spectra of Y at these wavelengths (FY) is calculated.
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