A novel rapid method for simultaneous determination of three diagnostically important porphyrins in erythrocytes using hyphenated synchronous fluorescence techniques

Liu, Qian; Huang, Wei; Shindi, Ali Abbas Falih; Li, Yao‐Qun

doi:10.1016/j.talanta.2011.11.057

Cited by 13 publications

(8 citation statements)

References 29 publications

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“…Protoporphyrin IX conforms to the common absorption characteristics reported in other tetrapyrroles with an absorbance maximum (λ abs ) at ~400 nm known as the Soret band (or B-band) which is complemented by smaller absorbance peaks at higher wavelengths (Q-bands) [ 5 ]. These absorption spectra are echoed in fluorescence spectrophotometry studies, which reported similar excitation maxima (λ ex ) with a subsequent fluorescence emission maxima (λ em ) ranging from 632.7 to 636 nm via the Q(0,0) transition [ 6 , 7 , 8 , 9 ]. The peak λ em for protoporphyrin IX occurs together with a second, smaller emission peak at ~700 nm via the Q(0,1) transition known as the J-band which is also associated with the result of the formation of J-aggregates at intermediate pH when the porphyrin molecules self-aggregate [ 10 , 11 ].…”

Section: Introductionmentioning

confidence: 81%

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A Preliminary Study of the Effects of pH upon Fluorescence in Suspensions of Prevotella intermedia

et al. 2016

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The quantification of fluorescence in dental plaque is currently being developed as a diagnostic tool to help inform and improve oral health. The oral anaerobe Prevotella intermedia exhibits red fluorescence due to the accumulation of porphyrins. pH affects the fluorescence of abiotic preparations of porphyrins caused by changes in speciation between monomers, higher aggregates and dimers, but this phenomenon has not been demonstrated in bacteria. Fluorescence spectra were obtained from suspensions of P. intermedia that were adjusted to pHs commensurate with the range found within dental plaque. Two fluorescent motifs were identified; 410 nm excitation / 634 nm emission (peak A) and 398 nm excitation / 622 nm emission (peak B). A transition in the fluorescence spectra was observed from peak A to peak B with increasing pH which was also evident as culture age increased from 24 hours to 96 hours. In addition to these ‘blue-shifts’, the intensity of peak A increased with pH whilst decreasing with culture age from 24 to 96 hours. A bacterium’s relationship with the local physiochemical environment at the time of image capture may therefore affect the quantification of dental plaque fluorescence.

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Section: Introductionmentioning

confidence: 81%

“…The fluorescent properties of coproporphyrin found in P . intermedia are λ ex 397 nm with λ em 623 nm [ 6 , 23 ].…”

Section: Introductionmentioning

confidence: 99%

A Preliminary Study of the Effects of pH upon Fluorescence in Suspensions of Prevotella intermedia

et al. 2016

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“…17,25,26 These studies suggested that porphyrins can undergo a transition from their monomeric form to higher aggregates and oligomers, particularly dimers, in response to the increasing pH. 27 The concentration of a porphyrin can also affect its fluorescence characteristics due to a very similar phenomenon.…”

Section: Discussionmentioning

confidence: 99%

Evaluating the effect of local pH on fluorescence emissions from oral bacteria of the genusPrevotella

Hope

Higham

2016

J. Biomed. Opt

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Abstract. A number of anaerobic oral bacteria, notably Prevotellaceae, exhibit red fluorescence when excited by short-wavelength visible light due to their accumulation of porphyrins, particularly protoporphyrin IX. pH affects the fluorescence of abiotic preparations of porphyrins due to transformations in speciation between monomers, higher aggregates, and dimers. To elucidate whether the porphyrin speciation phenomenon could be manifested within a microbiological system, suspensions of Prevotella intermedia and Prevotella nigrescens were examined by fluorescence spectrophotometry while being titrated against NaOH. The initial pH of the samples was <6, which was then raised toward the maximum found within a diseased periodontal pocket, being ∼pH 8.7. The intensity of the fluorescence emissions increased between 600 and 650 nm with increasing pH. Peak fluorescence emissions occurred at 635 AE 1 nm with a second emission peak developing with increasing pH at 622 nm. A linear relationship was demonstrated between pH and the log 10 ratio of 635:622 nm excitation fluorescence intensities. These findings suggest that the pH range found within the oral cavity could affect the fluorescence of oral bacteria in vivo, which may in turn have connotations for any clinical diagnoses that may be inferred from dental plaque fluorescence. © The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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“…To calculate the ZnPP/heme and PPIX/heme ratios, an additional measurement of the circulating hemoglobin concentration is required. A variety of other methods for determination of both ZnPP and PPIX have been developed [3,[23][24][25][26][27][28][29] but, for the most part, have proven too technically demanding or cumbersome for general adoption. For measurement of ZnPP alone, a portable front-face fluorometer, the hematofluorometer [30], designed for easy use, was introduced over three decades ago and remains commercially available (AVIV Hematofluorometer, Model 206D, Aviv Biomedical, Inc., Lakewood, NJ, USA).…”

Section: Introductionmentioning

confidence: 99%

Dual‐wavelength excitation for fluorescence‐based quantification of zinc protoporphyrin IX and protoporphyrin IX in whole blood

Hennig

Gruber

Vogeser

et al. 2013

Journal of Biophotonics

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Journal of BIOPHOTONICSQuantification of erythrocyte zinc protoporphyrin IX (ZnPP) and protoporphyrin IX (PPIX), individually or jointly, is useful for the diagnostic evaluation of iron deficiency, iron-restricted erythropoiesis, lead exposure, and porphyrias. A method for simultaneous quantification of ZnPP and PPIX in unwashed blood samples is described, using dual-wavelength excitation to effectively eliminate background fluorescence from other blood constituents. In blood samples from 35 subjects, the results of the dualwavelength excitation method and a reference high performance liquid chromatography (HPLC) assay were closely correlated both for ZnPP (r s ¼ 0.943, p < 0.0001; range 37-689 mmol ZnPP/mol heme, 84-1238 nmol/L) and for PPIX (r s ¼ 0.959, p < 0.0001; range 42-4212 mmol PPIX/mol heme, 93-5394 nmol/L). In addition, for ZnPP, the proposed method is compared with conventional single-wavelength excitation and with commercial front-face fluorimetry of washed erythrocytes and whole blood. We hypothesize that dual-wavelength excitation fluorimetry will provide a new approach to the suppression of background fluorescence in blood and tissue measurements of ZnPP and PPIX.

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A novel rapid method for simultaneous determination of three diagnostically important porphyrins in erythrocytes using hyphenated synchronous fluorescence techniques

Cited by 13 publications

References 29 publications

A Preliminary Study of the Effects of pH upon Fluorescence in Suspensions of Prevotella intermedia

A Preliminary Study of the Effects of pH upon Fluorescence in Suspensions of Prevotella intermedia

Evaluating the effect of local pH on fluorescence emissions from oral bacteria of the genusPrevotella

Dual‐wavelength excitation for fluorescence‐based quantification of zinc protoporphyrin IX and protoporphyrin IX in whole blood

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