Carboxyhemoglobin (COHb) quantitation is a routine toxicological analysis performed in cases of suspected acute carbon monoxide poisoning. Current methods available for COHb quantitation include spectrophotometric methods and gas chromatographic methods, both of which have advantages and disadvantages. The goal of the present study was to develop a Raman microspectrophotometric COHb quantitation method and assess its precision and accuracy. A strong linear correlation (average R² = 0.98) was found to exist between COHb concentration and the ratio of the Raman intensities at approximately 1,552 and 1,580 cm⁻¹. The limits of detection and quantitation were found to be 4.4% and 13.2% COHb, respectively. Intraday and interday studies of three calibrators indicated high precision, with the coefficient of variation ranging from 0.85% to 3.4%. The accuracy of this method was evaluated through the quantitation of blind samples, first using the proposed Raman microspectrophotometric method and then by a published UV–visible spectrophotometric method. For the proposed Raman method, 87.5% of the samples were quantitated with less than 20% error. The average variances of the two methods were found to be statistically different using a two‐sample F test, with the Raman microspectrophotometric method having a lower average variance. This study demonstrates that Raman microspectrophotometry may be used to quantitate the COHb concentration of blood samples. Furthermore, this method is comparable with established spectrophotometric methods while eliminating the need for time‐consuming sample preparation and limiting the use of consumables and the generation of waste.

中文翻译：

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拉曼光谱法测定血液中羧基血红蛋白的方法开发

碳氧血红蛋白（COHb）定量是在怀疑急性一氧化碳中毒的情况下进行的常规毒理学分析。目前可用于COHb定量的方法包括分光光度法和气相色谱法，两者都有优缺点。本研究的目的是开发一种拉曼显微分光光度法COHb定量方法，并评估其准确性和准确性。发现在大约1,552和1,580 cm ^-1的COHb浓度与拉曼强度比之间存在很强的线性相关性（平均R ² = 0.98）。。检出限和定量限分别为4.4％和13.2％COHb。对三个校准器的日间和日间研究表明精度很高，变异系数范围为0.85％至3.4％。该方法的准确性是通过对盲样品进行定量来评估的，首先使用提议的拉曼显微分光光度法，然后使用已公开的紫外可见分光光度法。对于拟议的拉曼方法，定量了87.5％的样品，误差小于20％。使用两样本F，发现两种方法的平均方差在统计学上不同拉曼显微分光光度法的平均方差较低。这项研究表明，拉曼显微分光光度法可用于定量血液样品的COHb浓度。此外，该方法可与既定的分光光度法相媲美，同时消除了耗时的样品制备需求，并限制了消耗品的使用和废物的产生。