RESEARCH Abstract Purpose: The aim of this study was to examine the accuracy of arterial oxygen saturation (Sao2) and venous oxygen saturation (Svo2) with paired arterial and venous (AV) blood in relation to pulse oximetry saturation (Spo2) and oxyhemoglobin (Hbo2) with fetal hemoglobin determination, and their Hbo2 dissociation curves. Method: Twelve preterm neonates with gestational ages ranging from 27 to 34 weeks at birth, who had umbilical AV lines inserted, were investigated. Analyses were performed with 37 pairs of AV blood samples by using a blood volume safety protocol. Results: The mean differences between Sao2 and Svo2, and AV Hbo2 were both 6 percent (F6.9 and F6.7 percent, respectively), with higher Svo2 than those reported for adults. Biases were 2.1 F 0.49 for Sao2, 2.0 F 0.44 for Svo2, and 3.1 F 0.45 for Spo2, compared against Hbo2. With left-shifted Hbo2 dissociation curves in neonates, for the critical values of oxygen tension values between 50 and 75 millimeters of mercury, Hbo2 ranged from 92 to 93.4 percent; Sao2 ranged from 94.5 to 95.7 percent; and Spo2 ranged from 93.7 to 96.3 percent (compared to 85 – 94 percent in healthy adults). Conclusions: In neonates, both left-shifted Hbo2 dissociation curve and lower AV differences of oxygen saturation measurements indicated low flow of oxygen to the body tissues. These findings demonstrate the importance of accurate assessment of oxygenation statues in neonates. n 2005 Elsevier Inc. All rights reserved. Accurate Measurements of Oxygen Saturation in Neonates: Paired Arterial and Venous Blood Analyses By Shyang-Yun Pamela K. Shiao, PhD, RN, FAAN O xygen saturation (So2) measurements (functional measurement, So2; and fractional measurement, oxyhemoglobin [Hbo2]) and monitoring are commonly investigated as a method of assessing oxygenation in neonates. Differences exist between the So2 and Hbo2 when blood tests are performed, and clinical monitors indicate So2 values.1 Oxyhemoglobin will decrease with the increased levels of carbon monoxide hemoglobin (Hbco) and methemoglobin (MetHb), and it is the most accurate measurements of oxygen (O2) association of hemoglobin (Hb). Pulse oximeter (for pulse oximetry saturation [Spo2] measurement) is commonly used in neonates. However, it will not detect the changes of Hb variations in the blood for accurate So2 measurements.2 – 4 Thus, the measurements from clinical oximeters should be used with caution. In neonates, fetal hemoglobin (HbF) accounts for most of the circulating Hb in their blood. Fetal hemoglobin has a high O2 affinity, thus releases less O2 to the body tissues, presenting a left-shifted Hbo2 dissociation curve.5,6 To date, however, limited data are available with HbF correction, for accurate arterial and venous (AV) So2 measurements (arterial oxygen saturation [Sao2] and venous oxygen saturation [Svo2]) in neonates, using paired AV blood samples. Fetal Hemoglobin and Oxygen Saturation Measurements From the School of Nursing, University of Texas Health Science Center at Houston, Houston, TX. Address reprint requests to Shyang-Yun Pamela Shiao, PhD, RN, FAAN, School of Nursing, University of Texas Health Science Center at Houston, 6901 Bertner Ave. SONSCC 567-8, Houston, TX 77030. n 2005 Elsevier Inc. All rights reserved. 1527-3369/05/0504-0112$30.00/0 doi:10.1053/j.nainr.2005.09.001 A ccurate measurements of So2 are dependent on HbF, Hbo2, and related parameters. Although HbF was useful to fetus to compete O2 at placenta with mother’s adult hemoglobin (HbA), it compromises oxygenation to the body tissue after birth. Neonates manifest desaturation events as the pulse oximeter loses sensitivity for measurement.7 A pulse oximeter can overestimate So2 by as much as 6 percent when HbF is not accounted for.7 –11 When low O2 Newborn and Infant Nursing Reviews, Vol 5, No 4 (December), 2005: pp 170 –178 170 Arterial and Venous Oxygen Saturation was delivered to the body tissue during events such as bradycardia and desaturation (b90 percent for Sao2), the neonate’s oxygenation would be further compromised. In previous reports, widely spread Spo2 readings had been associated with Pao2 values without providing a reasonably precise Hbo2 dissociation curve.12,13 The accuracy of pulse oximetry is limited when the saturation drops to lower than 80 percent,8,14,15 and its accuracy is of greater concern for neonates with HbF and low oxygen to the body.16 The normal clinical Pao 2 range was defined as 50 to 75 millimeters of mercury for infants and could be lower for preterm infants.17 In adults, Spo2 of 85 to 94 percent was associated with Pao2 of 50 to 75 millimeters of mercury.18 Comparable ranges of So2 need to be established for neonates, taking into the account of HbF determination. Arterial and Venous S O 2 O nly one previous study provided complete information on the validation of Sao2 and Svo2 in neonates, however, without HbF determination and related adjustments.19 Arterial oxygen saturation and venous oxygen saturation measurements together can be used to assess the systemic supply and demand of O2 for systemic oxygenation status. 20 –23 Venous oxygen saturation changes occur sooner and in more obvious increments than decreases in Sao2 during nursing care and interventions in adults.20 – 22 Venous oxygen saturation is rarely measured in neonates,24 perhaps with the concern for the accuracy of measurement for the presence of patent ductus arteriosus and patent foramen ovale. These features could cause central hemodynamic shunting resulting from persistent fetal circulation, thus compromising oxygenation status to the body tissue. Nonetheless, the hemodynamic shunting can be assessed when AV So2 is measured in neonates.23,24 Arterial and Venous Hb CO C arbon monoxide hemoglobin is a common cause to overestimate So2 in clinical monitoring when hemoximeter measurements are not determined using the blood analyses. In addition, when HbF was not corrected on the hemoximeter tests, Hbco readings were artificially increased, which then widened the differences between So2 and Hbo2 readings.4,5 The most common reason for elevated Hbco is inhalation of CO in fire accidents.3,25,26 Examination of AV Hbco in adults yielded high correlations (0.97 – 0.99) with a negligible mean AV Hbco difference of 0.15 percent.27,28 171 In a study of critically ill adult patients, increased pulmonary CO production and elevation in arterial Hbco but not venous Hbco were documented by inflammatory stimuli inducing pulmonary heme oxygenase–1.29 In normal adults, venous Hbco level might be slightly higher than or equal to arterial Hbco because of production of CO by enzyme heme oxygenase–2, which is predominantly produced in the liver and spleen. However, hypoxia or pulmonary inflammation could induce heme oxygenase–1 to increase endogenous CO, thus elevating pulmonary arterial and systemic arterial Hbco levels in adults.30 Both endogenous and exogenous CO can suppress proliferation of pulmonary smooth muscles, a significant consideration for the prevention of chronic lung diseases in newborns. Despite these considerations, a later study31 in healthy adults indicated that the AV differences in Hbco were from technical artifacts and perhaps from inadequate control of different instruments. Thus, further studies are needed to provide more definitive answers for the AV differences of Hbco for adults and neonates with acute and chronic lung diseases. Arterial and Venous MetHb M ethemoglobin is an indicator of Hb oxidation and is essential for accurate measurement of Hbo2, So2, and oxygenation status. No evidence exists to show the AV MetHb difference, although this difference was elucidated with the potential changes of MetHb with different O2 levels. Methemoglobin can be increased with nitric oxide (NO) therapy, used in respiratory distress syndrome (RDS) to reduce pulmonary hypertension and during heart surgery.32,33 Nitric oxide, in vitro, is an oxidant of Hb, with increased O2 during ischemia reperfusion. In hypoxemic conditions in vivo, nitrohemoglobin is a product generated by vessel responsiveness to nitrovasodilators.34 Nitro-hemoglobin can be spontaneously reversible in vivo, requiring no chemical agents or reductase. However, when O2 levels were increased experimentally in vitro following acidic conditions (pH 6.5) to simulate reperfusion conditions, MetHb levels were increased for the hemolysates (broken red cells).35 Nitrite-induced oxidation of Hb was associated with an increase in red blood cell membrane rigidity, thus contributing to Hb breakdown.35 A newer in vitro study of whole blood cells, however, concluded that MetHb formation is not dependent on increased O2 levels.36 Additional studies are needed to examine in vivo reperfusion of O2 and MetHb effects. For oxidation stress to the tissue and cells occurring with reperfusions, both animal models37,38 and human (newborn umbilical blood) studies39,40 have found DNA damage when oxygenation was increased after hypoxia 172 Shyang-Yun Pamela K. Shiao during perinatal asphyxia to the fetuses and neonates. Cellular oxidation injury might occur at various organs, including brain tissue, after hypoxia (low O2 to the tissue) in neonates.37 – 40 Considering that serum antioxidant activity is lower in neonates than adults, especially during the ischemia-reperfusion period,41 monitoring oxygenation status is critical. Electron carriers such as methylene blue as strong reducing agents can be used to treat elevated MetHb levels.32 These findings for MetHb also indicate the rationale to use fresh HbA for blood transfusion in neonates. In stored blood, as the Hb cells break down, MetHb levels could increase because of accelerated ferrous activity.42 Transfusing old HbA cells could impair cardiac function and coronary flow in neonatal hearts experiencing ischemia and reperfusion.41 The most recent recommendation for the maximum storage of blood for transfusion is 35 days43,44 because the half-life of transfused HbA was about 30 days.43 Further studies are needed for neonates with reperfusion and blood transfusion during ventilatory support. Arterial and venous MetHb differences could be helpful in assessing oxidation stress with changing oxygenation status in neonates. The purposes of this study, therefore, were (1) to examine the difference between So2 (Sao2, Svo2, and Spo2) in relation to Hbo2 measurements (accounting Hbco and MetHb) with HbF determination using paired AV blood samples and pulse oximetry, and (2) to present the Hbo2 dissociation curves in relation to O2 tension (Po2) values for accurate assessment of oxygenation in neonates. Comparing paired AV blood samples allows the examination of systemic balance of oxygenation status in neonates. Methods Setting This study is part of a larger clinical study involving around-the-clock data collection for neonates in three neonatal intensive care units. The appropriate institutional human subjects review boards approved the study protocols. Informed consents were obtained from the parents and guardians of all newborn subjects before or immediately after the births of high-risk neonates. As part of care for severe respiratory distress (RDS) and ventilatory support in neonatal intensive care units, umbilical artery catheters (UACs) and umbilical venous catheters (UVCs) were inserted for these neonates as central line access to assess blood O2 levels and to provide nutrients for the neonates. Umbilical artery catheters were inserted at the high positions (above lumbar 1) because lower placements were more likely to cause vascular spasm to the lower extremities. Umbilical venous catheters were inserted approximately 1 centimeter above the liver, at inferior vena cava. Blood was obtained through the UAC and UVC lines, from first to fifth days of life, coordinated to occur every 8 hours with routine blood gas tests. To conserve blood volume for these critically ill neonates, a safety blood volume protocol was instituted. Less than 4 milliliters per kilogram of blood was drawn from each subject during the entire duration of the study. Medical records were prospectively reviewed to obtain subject’s demographic data, medical history, and laboratory and monitoring parameters. Sample Immediately after birth, neonates who were diagnosed with RDS requiring ventilatory support were included in the study. All neonates had successful insertions of UAC and UVC. Neonates with major congenital defects (heart, brain, and neurological, or gastrointestinal defects) were excluded because interpretation of accurate Sao2 and Svo2 would be problematic in these cases. Neonates who had life-threatening persistent pulmonary hypertension and needed NO treatments or extracorporeal membrane oxygenation were excluded with added features to intensive care for feasibility concerns. A priori power analysis indicated that 36 (nonindependent) blood tests were needed for accuracy testing of Spo2 with HbF correction. The sample included 12 neonates, all having severe respiratory distress, who provided 37 pairs of AV blood samples. Gestational ages of the neonates ranged from 27 to 34 weeks, and birth weights ranged from 875 to 2245 grams. Instruments Fetal hemoglobin and all So2 parameters were measured by using a hemoximeter (co-oximeter) model OSM3 (Radiometer Corp, Cleveland, Ohio), using sixwavelength fiberoptic reflectance oximetry (535, 560, 577, 622, 636, and 670 nanometers). Newer models of hemoximeter (after 1993) have been reported to adjust So2 or Hbo2 readings by HbF levels using a linear relationship assumption.45 The accuracy of this newer cooximeter, as reported by the manufacturer, is a test-retest variability of less than 0.1 percent for normal Hb level and 0.2 to +0.4 percent for extreme anemia and polycythemia (Hb measurement ranges, 3.2 to 28 grams per deciliter). The instrument allowed in vitro measurements of So2, Hbo2, total hemoglobin (tHb) levels, and HbF concentrations through determination of P50 on the Hbo2 dissociation curve. Validity was ensured by zero-point calibration using the manufacturer’s rinse solution before Arterial and Venous Oxygen Saturation and after each test. Quality control procedures included the reference method every 8 hours, cleansing the internals of tubing with appropriate solutions every week, and changing maintenance tubing and tHb calibration every quarter to ensure test accuracy. To follow the recommended guidelines from a consensus meeting for So2 measurements,46 the cap for the restrictions of 100 percent maximum for So2 and Hbo2 measurements was lifted so that the test results exceeding 100 percent can be shown as measured by the equipment. Blood gas and monitor data were recorded simultaneously with the blood tests. Clinical monitor readings included pulse oximeter readings, respiratory rate and heart rate readings, and incubator temperature and skin temperature readings. Pulse oximetry saturation readings were recorded by using a pulse oximeter (Nellcor NPB 290; Tyco HealthCare, Mansfield, Mass) for all subjects. This instrument was capable of measuring the percentage of peripheral So2 detected transcutaneously by a probe positioned on either side of a pulsating arterial bed around the neonate’s foot. The transmittance sensor was configured so that the lightemitting diodes transmit infrared and red light through the pulsating vascular bed to a photodetector positioned on the opposite site.47 The sensors for pulse oximeter were all placed postductally on infants’ foot. The pulse oximeter has shown excellent correlations (r = 0.98 –0.99) with Sao2 without corrections for HbF percentage in neonates48,49 and sound correlations (r = 0.5, 0.88) with corrections for HbF.7,50 Interrater agreement on data coding was double-checked to reach 100 percent to ensure no difference was present between raters. Procedures Paired AV blood samples were obtained through UAC and UVC every 8 hours in coordination with routine blood gas sampling when neonates were sleeping quietly to obtain stable measurements. All clinical monitor readings of vital functions except respiratory rate were observed to ensure their normal ranges with the blood sampling. The mean readings of clinical monitors were Spo2 97.1 F 2.4 percent, 137 F 14.7 beats per minute for heart rate, 36.4 F 0.83 degrees Celsius for incubator temperature, 36.7 F 0.2 degrees Celsius for skin temperature, and 41.6 F 19.5 breaths per minutes for respiratory rate. The hemoximeter test, including HbF determination, needed less than 0.1 milliliter blood. As part of study protocol, blood samples were split for the HbF test and the So2 tests. Fetal hemoglobin determination included oxygenating blood samples with 100 percent O2 and twirling the syringe containing the sample between the hands for 90 seconds to yield fully oxygenated samples. 173 The oxygenation status was confirmed by So2 readings close to 100 percent. Fetal hemoglobin was then determined by using the hemoximeter. Using this determined HbF value, the blood So2 test was performed to determine So2, Hbo2, Hbco, MetHb, and reduced Hb (or deoxyhemoglobin) in paired AV blood samples when routine blood gas tests were performed. Data Analysis Data were examined with AV blood samples for matched HbF readings. Although the training protocol was used, some HbF determinations indicated inadequate oxygenation because technical errors were difficult to prevent for this 24-hour, around-the-clock study, particularly for newer staff personnel. Without adjusting HbF correctly, Hbco values were an average of 2 percent higher (r = 0.70, df = 73, P b .001), which caused significant differences between So2 and Hbo2 measurements.51 Thus, using a 4 percent criterion,20 unmatched HbF results between AV blood samples were not included in the analysis. The final sample included 37 pairs of AV blood samples (1–14 per subject). Although one neonate provided more samples than others, no difference was found when the data were compared with the subject from other subjects. Data were analyzed by using Statistical Packages for Social Studies (SPSS, Chicago, Ill). Multivariate linear mixed model (MLMM) approach51 (maximum likelihood method for the model estimation and autoregressive model for repeated measurements) was attempted to examine the differences between the measurements for repeated measurements of multiple data points from some subjects. When the levels of the repeated effects were not different for the observations within the repeated subjects to warrant the MLMM approach, the original technique of Bland and Altman52 was used. The MLMM approach51 was an addition to the original technique reported by Bland and Altman52,53 to test hypotheses using multivariate level analyses for repeated measurements. However, the original technique of Bland and Altman52 still provided additional useful information and interpretation such as bias and precision, and limits of agreement. The bias was defined as the mean difference, and the precision was defines as the SE of the mean difference. The limits of agreement as defined by Bland and Altman52 is a proportional function of distribution for differences between the two measurements. With normal distributions, 67 percent should fall within F1 SD of the mean difference and 95 percent should fall within F2 SD of the mean difference, based on the statistical principles. Oxygen saturation measurements along the Hbo2 dissociation curves need to be examined for the critical 174 Shyang-Yun Pamela K. Shiao values in clinical assessment. Oxyhemoglobin dissociations were examined by using multiple regression (R) curvefitting analysis on sigmoid (S) curve for Po2 values and all of the So2 measurements of the study (Hbo2, So2, and Spo2). These curves are useful when examining the relative values of So2 measurements (So2, Hbo2, and Spo2) against clinical diagnostic Po2 values for the detection of hypoxemia (b50 millimeters of mercury) and hyperoxemia (N75 millimeters of mercury). Results Table 2. Summary Statistics on Mean Difference, SE, and SDs for SO2 Against HbO2 With Arterial, Venous, and Total Blood Samples Parameters Arterial Venous Total Correlation Mean difference SE F1 SD Percent total F2 SD Percent total 0.96T 2.14T 0.08 F0.49 70.3 F0.97 97.3 1.00T 2.00T 0.07 F0.44 59.5 F0.88 97.3 1.00T 2.07T 0.05 F0.47 67.5 F0.93 97.2 TP b .001. T able 1 presents descriptive statistics for hemoximeter and blood gas analyzer results for general references and the differences between AV blood samples of these measurements. The mean difference between AV pairs on HbF was 0.3 F 1.7 percent and not significant (NS). The difference for AV Hbco was 0.03 F 0.13 percent (NS), and the correlation for paired AV Hbco was 0.96 ( P b .001). The mean difference for AV MetHb was 0.02 F 0.14 percent, and the correlation for paired AV MetHb was 0.52 ( P b .001). The limits of agreement with F2 SD for the disagreement between AV measurements were 5.4 percent for Hbco and 2.7 percent for MetHb. The difference for AV So2 was 6 F 7.91 percent ( P b .001) and the correlation was 0.04 (NS), whereas the difference for AV Hbo2 was 5.9 F 7.63 percent ( P b .001) and the correlation was 0.04. For limits of agreement with F2 SD, the disagreements between AV measurements were 2.7 percent (percent outside the 2 SD) for both So2 and Hbo2. The low differences between AV So2 (6 percent) indicated that these neonates had low oxygen to the body tissue. Carbon monoxide hemoglobin and methemoglobin, together, on average, accounted for less than 2.3 percent of total Hb for both AV blood samples (Table 1). Table 2 presents the differences between measurements for the accurate So2 measurements. The bias for So2 against Hbo2 was 2.14 F 0.49 percent for arterial samples, 2 F 0.44 percent for venous samples, and 2.07 F Table 1. Descriptive Statistics and Differences Between AV Blood Samples Arterial Venous Differences OSM3 hemoximeter tHb (grams per deciliter) HbF (percent) Hbco (percent) MetHb (percent) Hbo2 (percent) RHb (percent) So2 (percent) 12.9 94.1 1.6 0.7 93.7 4.1 95.8 F F F F F F F 2.24 20.55 0.47 0.14 1.72 1.68 1.72 12.9 93.8 1.5 0.7 87.8 10.0 88.9 F F F F F F F 2.28 20.97 0.44 0.14 7.60 7.60 7.83 0.2 0.3 0.03 0.02 5.9 5.9 6.0 Blood gas analyzer So2 (percent) pH Po2 (millimeters of mercury) Pco2 (millimeters of mercury) HCO3 (milliequivalents per liter) Base excess 94.2 7.4 99.9 35.3 20.8 3.4 F F F F F F 4.06 0.06 73.15 5.39 2.77 3.14 80.7 7.3 58.8 37.7 21.0 4.1 F F F F F F 12.71 0.09 42.40 5.38 4.55 5.28 13.5 F 0.03 F 41.5 F 2.3 F 0.5 F 1.0 F Values are presented as mean F SD. RHb indicates reduced Hb. TP b .001. TTP b .05. TTTP b .01. F F F F F F F 1.41 1.65 0.13 0.14 7.63T 7.76T 7.91T 13.57T 0.07TT 61.83T 1.65TTT 4.05 3.41 Arterial and Venous Oxygen Saturation Table 3. Summary Statistics on Mean Difference, SE, and SDs for SpO2 Against SO2 and HbO2 With Arterial Blood Samples 175 PO2 vHbO2 S-CURVE PO2 SvO2 S-CURVE 100 98 96 Parameters (percent) Spo2-So2 Spo2-Hbo2 Correlation Mean difference SE F1 SD Percent total F2 SD Percent total 0.44T 0.96TT 0.45 F2.74 70.3 F5.47 94.6 0.44T 3.10TTT 0.45 F2.73 73 F5.46 94.6 94 92 90 % 88 86 84 82 80 TP b .01. TTP b .05. TTTP b .001. 78 76 30 40 50 60 70 80 90 100 PvO2, mm Hg 0.47 percent for total AV blood samples (all P b .001). The correlation between So2 and Hbo2 was close to 1.0 for all samples, 0.96 for arterial samples, 1.0 for venous samples, and 1.0 for total blood samples (all P b .001). For the limits of agreement within F2 SD, the disagreements between So2 and Hbo2 were 2.7 percent for both paired AV samples and 2.8 percent for total samples. The same analyses were performed for Spo2 and related measurements. The biases for Spo2 against Sao2 and arterial Hbo2 were 1 percent ( P b .05) and 3.1 percent ( P b .001), respectively (Table 3). Therefore, the mean bias for pulse oximeter readings against the most accurate mea- PO2 aHbO2 SCURVE PO2 SaO2 S-CURVE PO2 SpO2 S-CURVE 100 99 98 97 96 % 95 94 93 92 91 90 50 60 70 80 90 100 PaO2, mm Hg Fig 1. Arterial Hbo2 sigmoid curves with Pao2 for arterial Hbo2, Sao2, and Spo2. Fig 2. Venous Hbo2 sigmoid curves with Pvo2 for venous Hbo2 and Svo2. surement using arterial Hbo2 was 3.1 percent. The correlation between Spo2 and Sao2 was 0.44, and that between Spo2 and arterial Hbo2 was 0.44 (all P b .01). For limits of agreement, the disagreements were both 5.4 percent between Spo2 and Sao2, as well as for Spo2 and arterial Hbo2. Fig 1 presents the significant So2 dissociation sigmoid (S) fitted curves for arterial blood samples using multiple regression curve-fitting approach. For the ease of examinations, the ranges of Pao2 were focused at 100 millimeters of mercury or less. Significant S curves were established for Hbo2, Sao2, and Spo2 with Pao2. On the Hbo2 curves, in relation to Po2 ranges of 50 to 75 millimeters of mercury, arterial Hbo2 ranged from 92 to 93.4 percent (R = 0.514, P b .005), Sao2 ranged from 94.5 to 95.7 percent (R = 0.434, P b .01), and Spo2 ranged from 93.7 to 96.3 percent (R = 0.496, P b .01). Fig 2 presents the significant S curves for venous blood samples, and Fig 3 presents the significant S curves for the total merged blood samples. In Fig 2, for venous blood samples, in relation to venous oxygen tension (Pvo2) ranges of 50 to 75 millimeters of mercury, venous Hbo2 ranged from 88 to 94 percent (R = 0.514, P b .005) and Svo2 ranged from 90 to 96.2 percent (R = 0.801, P b .0001). The best S curves were produced when both AV blood samples were merged as the total samples, with greater R and better fit. In Fig 3, for the total merged blood samples, in relation to Po2 ranges of 50 to 75 millimeters of mercury, Hbo2 ranged from 88 to 93 percent (R = 0.807, P b .0001) and So2 ranged from 90 to 95 percent (R = 0.802, P b .0001). No significant S curves were established for Spo2 and Pvo2 176 Shyang-Yun Pamela K. Shiao PO2 HbO2 S-CURVE PO2 SO2 S-CURVE 100 98 96 94 92 90 % 88 86 84 82 80 78 76 30 40 50 60 70 80 90 100 PO2, mm Hg Fig 3. Oxyhemoglobin sigmoid curves for total merged AV blood samples with Po2 for Hbo2 and So2. and for Spo2 and Po2 with the total merged blood samples. No significant correlations were found between Spo2 and Svo2, or Spo2 and venous Hbo2. Discussion I n these neonates, the mean AV blood differences for both So2 and Hbo2 were about 6 percent, which was much lower than those reported for healthy adults (23 percent) for O2 supply and demand.20-23 In addition, with very high levels of HbF releasing less O2 to the body tissue, the results of blood analyses are worrisome for these critically ill neonates for low systemic oxygen states. O’Connor and Hall19 determined AV So2 in neonates without HbF determination. Much of the AV So2 difference is dependent on Svo2 measurement. The ranges of Svo2 spanned for 35 percent, and the ranges of Sao2 spanned 6 percent in these neonates (Fig 1). The greater intervals for Svo2 measurements contribute to greater sensitivity for the measurements (than Sao2 measurements) in responding to nursing care and changes of O2 demand.20,21 Thus, Svo2 measurement is essential for better assessment of oxygenation status in neonates. When neonates were resting quietly, the bias between So2 and Hbo2 was less than 2.5 percent on average with the correction of HbF factor, with another 1 percent difference between Spo2 and So2, and slightly greater than 3 percent between Spo2 and Hbo2 measurements. These findings were comparable to those reported for adults.54 However, blood sampling (either venous or arterial or both) is needed to accurately determine So2 because clinical monitors cannot detect changes of Hb variations for accurate So2 measurements.1,46,54 In summary, the mean difference between pulse oximetry and blood Hbo2 differed by 3.1 percent. The differences between the measurements could become greater as neonates develop desaturation events.7 The paired AV blood samples are difficult to obtain, especially when ensuring safety concerns for blood volumes in these vulnerable neonates, with accurate measurements. These delicate blood analyses require dedications of conscientious staff personnel. Repeated practices using simulated solutions and volunteer adult blood1 before handling neonates’ blood yielded the precision limits better than 0.1 percent limit recommended by the manufacturer’s standards, indicating successful training of staff personnel, excluding unmatched AV pairs. Precision (SE) is a common indicator of accuracy for instruments in marketing by manufacturers. The precision for clinical Spo2 against both So2 and Hbo2 was 0.45 percent, which was at least five times less precise than the blood analyses. These findings confirmed that clinical pulse oximeters cannot demonstrate the same precision standards as the benchtop instruments.54 On the sigmoid curves, the ranges of So2 readings in relation to Pao2 were much narrower for these neonates, when compared with that for adults. As compared with the ranges of 95 to 84 percent Spo2 in adults,18,47 for the Pao2 ranges from 50 to 75 millimeters of mercury, the ranges of So2 measurements in neonates were higher and narrower (about 94 to 96.3 percent for Spo2, 94.5 to 95 percent for Sao2, and 92 to 93.4 percent for arterial Hbo2). In relation to 50 millimeters of mercury of Pao2 measurement on the Hbo2 dissociation curve, critical So2 ranges of 92 to 94 percent were identified for neonates. With the limited accuracy capacity of pulse oximetry in neonates during desaturation events, it is preferable to keep the Sao2 and Spo2 measurements at greater than 92 to 94 percent. These narrower ranges of So2 measurements require clinicians to exercise more careful and closer observations to assess critical changes of oxygenation conditions in neonates. The findings of this study on AV differences of So2 were limited with very small number of paired AV blood samples. However, critically ill neonates need accurate assessment of oxygenation status because of HbF, which releases less O2 to the tissues. Decreased differences of AV So2 measurements added further possibilities of lower flow of O2 to the body tissues and demonstrated the greater need to accurately assess the proper oxygenation in the neonates. The findings of this study continued to Arterial and Venous Oxygen Saturation clarify the accuracy of So2 measurements for neonates. Additional studies are needed to examine So2 levels in neonates to further validate these findings by using larger sample sizes. Acknowledgments T hree different awards support this study in part: KCIAACN Critical Care Research Award from the American Association of Critical Care, Aliso Viejo, CA; Research Award from the National Association of Neonatal Nurses, Glenview, IL; and the National Institutes of Health, R01-NR04447, Bethesda, MD. The author acknowledges the nurses, physicians, and respiratory therapists who participated and helped with the blood sample collection at the clinical settings. References 1. Shiao S-YPK: Functional versus fractional oxygen saturation readings: bias and agreement using simulated solutions and adult blood. Biol Res Nur 3:210 – 221, 2002 2. Harris AP, Sendak MJ, Donham RT, et al: Absorption characteristics of human fetal hemoglobin at wavelengths used in pulse oximetry. J Clin Monit 4:175 – 177, 1988 3. Moyle JT: Uses and abuses of pulse oximetry. Arch Dis Child 74:77 – 80, 1996 4. Rausch-Madison S, Mohsenifar Z: Methodologic problems encountered with cooximetry in methemoglobinemia. Am J Med Sci 314:203 – 206, 1997 5. Wimberly PD: Oxygen monitoring in the newborn. Scand J Clin Lab Invest 127 – 130, 1993 (54 Suppl 214) 6. 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