Appendices Reference Values in Infancy and Childhood

APPENDIX 1

Hematologic Values * in Normal Fetuses at Different Gestational Ages

Week of Gestation	Hemoglobin (g/dL)	RBCs (×10 ⁶/mL)	Hematocrit (%)	Mean Corpuscular Volume (fL)	Total WBCs (×10 ⁶/µL	Corrected WBCs (×10 ⁶/µL)	Platelets (×10 ⁶/µL)
18-21 (N = 760)	11.69 ± 1.27	2.85 ± 0.36	37.3 ± 4.32	131.1 ± 11.0	4.68 ± 2.96	2.57 ± 0.42	234 ± 57
22-25 (N = 1200)	12.2 ± 1.6	3.09 ± 0.34	38.59 ± 3.94	125.1 ± 7.8	4.72 ± 2.82	3.73 ± 2.17	247 ± 59
26-29 (N = 460)	12.91 ± 1.38	3.46 ± 0.41	40.88 ± 4.4	118.5 ± 8.0	5.16 ± 2.53	4.08 ± 0.84	242 ± 69
>30 (N = 440)	13.64 ± 2.21	3.82 ± 0.64	43.55 ± 7.2	114.4 ± 9.3	7.71 ± 4.99	6.4 ± 2.99	232 ± 87

RBCs, Red blood cells; WBCs, white blood cells.

Modified from Forestier F, Daffos F, Catherine N, et al: Developmental hematopoiesis in normal human fetal blood. Blood 77:2360, 1991.

* Hematologic data obtained with a Coulter S plus II instrument. Total WBC count included nucleated red blood cells. Corrected WBC count included only WBCs, after subtracting the nucleated red cell component, based on a 100-cell manual differential.

APPENDIX 2

WBC Manual Differential Counts in Normal Fetuses at Different Gestational Ages

Week of Gestation	Lymphocytes (%)	Neutrophils (%)	Eosinophils (%)	Basophils (%)	Monocytes (%)	Nucleated RBCs (% of WBCs)
18-21 (N = 186)	88 ± 7	6 ± 4	2 ± 3	0.5 ± 1	3.5 ± 2	45 ± 86
22-25 (N = 230)	87 ± 6	6.5 ± 3.5	3 ± 3	0.5 ± 1	3.5 ± 2.5	21 ± 23
26-29 (N = 144)	85 ± 6	8.5 ± 4	4 ± 3	0.5 ± 1	3.5 ± 2.5	21 ± 67
>30 (N = 172)	68.5 ± 15	23 ± 15	5 ± 3	0.5 ± 1	3.5 ± 2	17 ± 40

RBCs, Red blood cells; WBCs, white blood cells.

From Forestier F, Daffos F, Catherine N, et al: Developmental hematopoiesis in normal human fetal blood. Blood 77:2360, 1991.

APPENDIX 3

Hematologic Values for Cord Blood (Vaginal Delivery and Cesarean Section * )

Characteristic	Study Sample (N = 167)		Vaginal Delivery (N = 63)		Cesarean Section (N = 104)		P-value ²
Characteristic	Median	Range	Median	Range	Median	Range	P-value ²
WBC (×10 ⁹per l)	15.1	5.54-39.7	18.4	12.0-34.1	13.6	8.54-39.7	<0.0001
RBC (×10 ¹²per l)	4.7	3.46-6.62	4.78	3.89-6.30	4.62	3.46-6.62	NS
Hb (g l ⁻¹)	174	130-234	176	140-230	171	130-234	NS
Hct (%)	53.6	40.1-73.1	54.7	41.9-73.1	52.6	40.1-72.2	NS
MCV (fl)	112	97.7-127	114	105-127	112	97.7-125	NS
MCH (pg)	36.5	31.4-41	36.5	31.4-41	36.6	32-39.9	NS
MCHC (g l ⁻¹)	324	303-359	323	308-359	324	303-344	NS
RDW (%)	17.4	14.2-23.6	17.4	14.9-23.6	17.4	14.2-23.3	NS
PLT (×10 ⁹per l)	270	161-607	297	169-607	254	161-424	0.0053
MPV (fl)	8.7	7.5-11.5	8.7	7.7-11.4	8.8	7.5-11.5	NS
Plateletcrit (%)	0.24	0.15-0.48	0.26	0.15-0.48	0.23	0.15-0.36	0.0056
CD34+ cells (×10 ⁶per l)	43.9	7.14-253	47.7	15.9-253	39.9	7.14-120	0.007

Hb, Hemoglobin; Hct, hematocrit; MCH, mean corpuscular hemoglobin; MCHC, mean corpuscular hemoglobin concentration; MCV, mean corpuscular volume; MPV, mean platelet volume; NS, not significant; plateletcrit, MPV × PLT; PLT, platelet; RBC, red blood cell; RDW, red blood cell distribution width; WBC, white blood cell.

From Eskola M, Juutistenaho S, Aranko K, et al: J Perinatol 2011; 258–262. Data obtained with Sysmex K-1000 analyzer (Sysmex, Kobe Japan).

* aP-values of the differences between vaginal delivery and cesarean section. The concentrations were standardized to exclude the varying effect of the anticoagulant.

APPENDIX 4

Hematologic Values * for Normal Cord Blood

	Mean ± SD
Red Blood Cells
Hb (g/dL)	15.3 ± 1.3
Hct (%)	49 ± 5
RBC (×10 ⁶/µL)	4.3 ± 0.4
MCV (fL)	112 ± 6
MCH (pg)	36.2 ± 2.2
MCHC (g/dL)	30.9 ± 1.3
CHCM (g/dL)	30.4 ± 1.2
% HYPO (MCHC <28 g/dL)	17.3 ± 11.9
% HYPER (MCHC >41 g/dL)	0.6 ± 0.3
% MICRO (MCV <61 fL)	0.8 ± 0.3
% MACRO (MCV >120 fL)	31.8 ± 9.7
Reticulocytes
%	3.63 ± 1.11
Absolute reticulocytes (×10 ⁹/L)	156.1 ± 47.7
MCVr (fL)	125.8 ± 7.3
CHCMr (g/dL)	25.6 ± 1.2
CHr (pg)	31.3 ± 1.4

CHCM, Cell hemoglobin concentration mean; CHCMr, reticulocyte cell hemoglobin concentration mean; CHr, reticulocyte hemoglobin content; Hb, hemoglobin; Hct, hematocrit; % HYPER, % hyperchromic red cells; % HYPO, % hypochromic red cells; % MACRO, % macrocytic red cells; MCH, mean corpuscular hemoglobin; MCHC, mean corpuscular hemoglobin concentration; MCV, mean corpuscular volume; MCVr, reticulocyte mean corpuscular volume; % MICRO, microcytic red cells; RBC, red blood cell; SD, standard deviation.

Modified from Diagne I, Archambeaud MP, Diallo D, et al: Parametres erythrocytaires et reservés en fer dans le sang du cordon. Arch Fr Pediatr 2:208, 1995; and from G Tchernia, personal communication. Data obtained in 142 specimens with H*2 Technicon analyzer.

* Values obtained with Technicon H*2 and H*3 Hematology analyzers (Bayer Diagnostics) in neonates delivered at term with weight ≥2500 g.

APPENDIX 5

Red Cell and Reticulocyte Indices, Serum Iron Status Markers in Cord Blood

Parameter	Mean	SD	Reference Range *
Blood Count and Cellular Indices
Hb, g/L	159	15	146-189
HCT, L/L	0.49	0.05	0.44-0.58
MCV, fL	109	4	102-118
MCVr, fL	124	6	115-136
MCH, pg	35	1	33-38
MCHC, g/L	325	10	306-342
%Retic, %	4.0	0.8	2.6-5.4
IRF-H, %	24.1	7.8	10.2-40.0
CHm, pg	34.9	1.3	32.5-37.2
CHr, pg	35.6	1.3	33.1-38.6
%HYPOm, %	3.0	3.0	0.4-9.9
%HYPOr, %	42.0	15.6	18.3-76.8
Serum Measurements
TfR, mg/L	2.0	0.7	1.2-4.0
Ferritin, µg/L	198	137	45-636
TfR-F index	0.95	0.43	0.49-2.1
Iron, µmol/L	27.4	7.7	12.2-42.1
Transferrin, g/L	2.0	0.4	1.2-2.9
TfSat, %	55	19	21-111

CHm, Cellular hemoglobin in red blood cells; CHr, cellular hemoglobin in reticulocytes; Hb, hemoglobin; HCT, hematocrit; %HYPOm, percentage of hypochromic red blood cells; %HYPOr, percentage of hypochromic reticulocytes; IRF-H, high immature reticulocyte fraction; MCH, mean cell hemoglobin; MCHC, mean cell hemoglobin concentration; MCV, mean cell volume of red blood cells; MCVr, mean cell volume of reticulocytes; %Retic, proportion of reticulocytes; TfR, transferrin receptor; TfR-F index, transferrin receptor/log (ferritin); TfSat, transferrin saturation.

From Ervasti M, Kotisaari S, Sankilampi U, et al: The relationship between red blood cell and reticulocyte indices and serum markers of iron status in the cord blood of newborns. Clin Chem Lab Med 45:1000–1003, 2007. Hematologic data obtained in 199 full-term newborn infants with a ADVIA 120 analyzer (Siemens Diagnostic Solutions, Terrytown, NY).

* For reference range calculations, only samples in which Hb was greater than 146 g/L were included.

APPENDIX 6

Erythroblast and Leukocyte Counts in Umbilical Cord Blood *

Type of Delivery	Erythroblast Count (×10 ⁹/L)	Leukocyte Count (×10 ⁹/L)
Spontaneous, vaginal (N = 55)	0.75 (0.0-5.3)	13.8 (7.25-48.0)
Elective cesarean section (N = 39)	0.30 (0.0-0.49)	10.6 (6.2-17.7)
Emergency cesarean section (N = 55)	1.10 (0.0-15.9)	13.5 (4.2-40.3)

From Thilaganthan B, Athansasiou S, Ozmen S, et al: Umbilical cord blood erythroblast count as an index of intrauterine hypoxia. Arch Dis Child 70:F192, 1994.

* Values are expressed as mean (range). Erythroblast counts were significantly higher in the spontaneous vaginal and emergency cesarean section groups compared with the elective cesarean group.

APPENDIX 10

Reference Ranges for Hb, HCt, Reticulocyte Parameters, and S-TfR During the First 15 Weeks of Life

Age, Week	Hb			Hct			Reticulocyte			CHr			IRF			S-TfR
		Reference Interval			Reference Interval			Reference Interval			Reference Interval			Reference Interval			Reference Interval
	mean	lower	upper	mean	lower	upper	mean	lower	upper	mean	lower	upper	mean	lower	upper	mean	lower	upper
0.5	150.5	101.8	222.4	0.45	0.30	0.65				35.7	31.5	39.9	36.6	13.5	59.7
1.5	138.7	97.9	196.3	0.41	0.29	0.58	59.5	20.2	175.6	35.0	31.1	38.9	35.5	13.7	57.2	1.4	0.9	2.4
2.5	128.8	94.0	176.5	0.38	0.28	0.52	66.8	24.7	181.1	34.3	30.6	38.1	34.3	13.6	55.1	1.3	0.8	2.3
3.5	120.6	90.0	161.5	0.36	0.27	0.47	74.1	29.2	187.9	33.7	30.1	37.4	33.3	13.1	53.4	1.3	0.8	2.1
4.5	113.8	86.2	150.3	0.34	0.26	0.44	81.2	33.7	195.8	33.1	29.5	36.7	32.2	12.4	52.1	1.2	0.7	2.0
5.5	108.3	82.5	142.1	0.32	0.25	0.42	87.9	37.8	204.3	32.6	29.0	36.2	31.3	11.6	50.9	1.2	0.7	1.9
7.0	101.9	77.8	133.7	0.30	0.23	0.39	96.7	43.1	216.7	31.9	28.3	35.5	29.9	10.3	49.6	1.1	0.7	1.8
9.0	96.9	73.5	127.7	0.29	0.22	0.38	105.2	48.4	228.8	31.2	27.5	34.8	28.4	8.6	48.1	1.1	0.7	1.8
11.0	94.9	71.7	125.7	0.28	0.21	0.37	109.0	51.3	231.9	30.6	26.9	34.3	27.0	7.4	46.7	1.1	0.7	1.9
13.0	96.1	72.6	127.3	0.28	0.21	0.37	107.6	51.4	225.2	30.2	26.5	33.9	25.9	6.7	45.0	1.2	0.7	2.0
15.0	100.5	75.9	133.1	0.30	0.22	0.39	101.2	48.1	212.9	30.0	26.4	33.6	24.9	6.7	43.3	1.3	0.8	2.2

CHr, Reticulocyte hemoglobin content; Hb, hemoglobin; Hct, hematocrit; IRF, immature reticulocyte fraction; S-Tfr, soluble transferrin receptor.

From Mäkela E, Takal TI, Suomine P, et al: Hematological parameters in preterm infants from birth to 16 weeks of age with reference to iron balance. Clin Chem Lab Med 46:551–557, 2008. Hematologic data obtained with ADVIA 120, Siemens Medical Solutions, Tarrytown, NY. S-TfR measured with an automated immunoturbidimetric methos (IDeA sTfR-IT, Orion Diagnostica), Espoo, Finland. Ferritin assayed with Elecsys ferritin electrochemiluminescence immunoassay on a Modular E analyzer (Roche Diagnostics).

APPENDIX 11

Reference Ranges for Ferritin During the First 15 Weeks of Life

Age, Week	Ferritin *			Ferritin, No Transfusions *
		Reference Interval			Reference Interval
	mean	lower	upper	mean	lower	upper
1.5	221.4	77.0	636.8	215.5	73.5	631.6
2.5	199.9	65.6	609.2	178.5	56.9	559.8
3.5	180.4	52.8	616.5	149.2	43.9	507.2
4.5	162.8	41.4	640.2	125.9	34.1	465.4
5.5	147.0	32.3	668.4	107.2	26.8	429.7
7.0	126.0	22.6	702.3	85.7	19.2	382.1
9.0	102.6	14.9	708.7	65.7	13.3	324.6
11.0	83.5	10.6	659.1	52.3	10.1	271.7
13.0	68.0	8.2	563.0	43.2	8.3	224.0
15.0	55.3	6.9	444.8	37.0	7.5	182.9

From Mäkela E, Takal TI, Suomine P, et al: Hematologic parameters in preterm infants from birth to 16 weeks of age with reference to iron balance. Clin Chem Lab Med 46:551–557, 2008. Ferritin assayed with Elecsys ferritin electrochemiluminescence immunoassay on a Modular E analyzer (Roche Diagnostics).

* The information under the column “Ferritin” shows values calculated for the neonates who had not been transfused during the preceding 2 weeks of measurements. The information under the column “Ferritin, no transfusions” shows the values calculated for those neonates who had not been transfused at all (indicating ferritin level development in the most stable preterm neonates).

APPENDIX 12

Reference Ranges for CBC, Reticulocytes, Ferritin, and S-TfR During the First Year of Life in Preterm and Full-Term Infants

Age, Weeks	Hb, g/L		Hct		RBC count, × 10 ¹²/L		MCV, fL		MCH, pg		Retic, × 10 ⁹/L
Age, Weeks	Mean	95% RI	Mean	95% RI	Mean	95% RI	Mean	95% RI	Mean	95% RI	Mean	95% RI
Preterm
20	112.2	92.0-136.8	0.327	0.271-0.395	4.03	3.15-4.92	81.6	73.2-90.0	28.1	25.2-31.0	77.4	39.0-153.5
25	116.8	98.7-138.1	0.340	0.287-0.401	4.27	3.48-5.07	80.1	72.3-88.0	27.7	24.8-30.5	69.7	36.7-132.5
30	120.8	103.6-140.9	0.351	0.300-0.411	4.47	3.70-5.25	79.0	71.2-86.7	27.3	24.4-30.3	64.1	34.3-119.6
35	124.3	106.7-144.7	0.360	0.309-0.421	4.64	3.83-5.44	78.1	70.1-86.0	27.1	24.0-30.2	60.1	32.4-111.8
40	127.0	108.5-148.6	0.368	0.315-0.431	4.76	3.92-5.60	77.5	69.3-85.7	26.9	23.7-30.1	57.6	30.9-107.2
45	129.0	109.6-151.8	0.374	0.319-0.438	4.85	3.98-5.71	77.3	68.9-85.6	26.8	23.5-30.0	56.3	30.1-105.1
50	130.2	110.3-153.6	0.378	0.323-0.442	4.89	4.03-5.75	77.3	69.0-85.6	26.7	23.6-29.9	56.1	30.0-105.2
55	130.6	111.1-153.6	0.380	0.326-0.442	4.90	4.07-5.73	77.6	69.6-85.7	26.8	23.7-29.8	57.2	30.3-107.7
60	130.2	112.0-151.3	0.379	0.329-0.437	4.87	4.11-5.62	78.2	70.7-85.8	26.9	24.0-29.7	59.4	31.1-113.5
Full-Term
20	120.0	102.2-140.9	0.345	0.293-0.407	4.32	3.76-4.87	80.3	74.2-86.4	27.9	25.6-30.2	45.5	25.3-82.2
25	118.9	101.9-138.8	0.345	0.296-0.402	4.38	3.84-4.92	79.0	73.1-84.9	27.3	24.9-29.6	45.2	25.1-81.5
30	118.3	101.5-137.7	0.345	0.298-0.400	4.44	3.90-4.98	78.1	72.3-83.9	26.8	24.3-29.2	45.4	26.1-78.9
35	118.0	101.5-137.3	0.346	0.300-0.400	4.48	3.93-5.03	77.6	71.8-83.4	26.5	23.9-29.0	46.0	27.6-76.6
40	118.2	102.0-137.1	0.348	0.303-0.400	4.51	3.96-5.06	77.4	71.6-83.2	26.3	23.7-28.9	47.1	28.8-76.8
45	118.9	102.9-137.3	0.351	0.307-0.401	4.54	3.99-5.08	77.6	71.7-83.5	26.3	23.6-29.0	48.7	28.7-82.5
50	120.0	104.2-138.1	0.354	0.313-0.402	4.55	4.01-5.08	78.2	72.2-84.1	26.5	23.8-29.2	50.9	26.9-96.3
55	121.5	105.4-140.0	0.359	0.319-0.403	4.55	4.04-5.06	79.1	73.1-85.1	26.9	24.2-29.6	53.7	23.9-121.0
Age, Weeks	Ferritin, µg/L		S-TfR, mg/L		CHr, pg		WBC count, × 10 ⁹/L		Platelet count, × 10 ⁹/L
Age, Weeks	Mean	95% RI	Mean	95% RI	Mean	95% RI	Mean	95% RI	Mean	95% RI
Preterm
20	37.8	7.37-193.9	1.52	1.07-2.16	29.8	26.6-33.1	9.48	5.63-16.0	478	294-777
25	30.1	7.48-120.8	1.54	1.10-2.15	29.6	26.4-32.8	9.60	5.73-16.1	465	290-744
30	24.7	7.06-86.6	1.56	1.12-2.16	29.4	26.2-32.7	9.71	5.82-16.2	452	286-714
35	21.1	6.41-69.5	1.57	1.14-2.17	29.3	26.0-32.6	9.83	5.92-16.3	439	280-687
40	18.6	5.80-59.9	1.59	1.16-2.19	29.2	25.9-32.6	9.94	6.01-16.5	427	274-663
45	17.1	5.41-53.8	1.61	1.17-2.21	29.2	25.8-32.6	10.1	6.09-16.6	415	268-642
50	16.2	5.25-49.8	1.63	1.18-2.25	29.3	25.8-32.7	10.2	6.17-16.8	403	260-623
55	15.9	5.31-47.5	1.65	1.18-2.30	29.4	25.8-32.9	10.3	6.24-17.0	391	252-607
60	16.2	5.44-47.9	1.67	1.18-2.36	29.5	25.9-33.2	10.4	6.31-17.2	380	244-593
Full-Term
20	71.7	21.5-239.7	1.49	1.06-2.08	29.8	27.4-32.1	9.36	5.84-15.0	426	282-646
25	51.1	16.1-162.2	1.52	1.08-2.12	29.2	25.7-32.6	9.19	5.73-14.7	415	269-640
30	38.5	11.5-128.7	1.54	1.09-2.19	28.8	24.7-32.9	9.02	5.57-14.6	404	258-634
35	30.6	8.76-107.0	1.57	1.09-2.27	28.7	24.5-32.9	8.85	5.37-14.6	394	247-628
40	25.8	7.46-89.1	1.60	1.08-2.37	28.9	25.1-32.6	8.68	5.14-14.7	383	237-621
45	22.9	7.01-75.0	1.63	1.07-2.48	29.2	26.4-32.1	8.52	4.88-14.9	373	227-614
50	21.6	6.78-68.6	1.66	1.05-2.62	29.9	27.6-32.2	8.36	4.61-15.2	363	218-607
55	21.5	5.90-78.0	1.69	1.04-2.76	30.8	27.0-34.5	8.21	4.33-15.5	354	209-599

CBC, Complete blood count; CHr, reticulocyte hemoglobin content; Hb, hemoglobin; Hct, hematocrit; MCH, mean corpuscular hemoglobin; MCV, mean corpuscular volume; RBC, red blood cell; Retic, reticulocyte; RI, reference interval; S-Tfr, soluble transferrin receptor; WBC, white blood cell.

From Takale TI, Mäkela E, Suomine P, et al: Blood Cell and iron status analytes of preterm and full-term infants from 20 weeks onward during the first year of life. Clin Chem Lab Med 48:1295–1301, 2010. Hematologic data obtained with ADVIA 120, Siemens Medical Solutions, Tarrytown, NY. S-TfR measured with an automated immunoturbidimetric methos (IDeA sTfR-IT, Orion Diagnostica), Espoo, Finland. Ferritin assayed with Elecsys ferritin electrochemiluminescence immunoassay on a Modular E analyzer (Roche Diagnostics).

APPENDIX 16

Reference Ranges for WBC, Hb, Hct, and MCV in African-American Children and Adolescents *

		African-American [Hb (g/dL)]		African-American [Hct (%)]		African-American [MCV (fl)]
Age (years)	African American (mean WBC)	Mean	−2 STD	Mean	−2 STD	Mean	−2 STD
2-5	7.33	11.94	10.37	35.98	31.54	79.81	74.65
6-10	6.74	12.44	10.3	37.27	32.71	81.44	76.45
11-15 male	6.32	13.2	10.97	39.29	32.91	83.20	77.99
11-15 female	6.58	12.67	10.1	37.95	33.41	83.60	78.01
16-18 male	5.91	14.35	11.30	43.50	37.62	86.02	80.85
16-18 female	6.92	12.55	10.27	37.39	31.87	85.26	79.43

Hb, Hemoglobin; Hct, hematocrit; MCV, mean corpuscular volume; STD, standard deviation; WBC, white blood cell.

From Robins EB, Blum S: Hematologic reference values for African American children and adolescents. Am J Hematol 82:611–614, 2007.

* Sample size is 5039. Mean values obtained from NHANES (National Health and Nutrition Examination Survey) and Robins studies.

APPENDIX 17

WBC and ANC Mean Values and Reference Intervals *

	12-18 Years: WBC		>18 Years: WBC		12-18 Years: ANC		>18 Years: ANC
	N		N		N		N
NHBM	401	3.2-9.3 (5.6)	547	3.1-9.9 (6.1)	401	1.0-6.2 (2.9)	547	1.3-6.6 (3.1)
MAM	493	4.1-11.4 (6.9)	776	4.1-11.4 (7.1)	492	1.8-8.2 (4.1)	776	2.1-7.9 (4.2)
NHWM	342	4.1-11.4 (6.8)	1473	4.1-11.8 (7.0)	342	1.8-7.2 (3.8)	1471	2.1-8.0 (4.3)
MAM, NHWM	835	4.1-11.4 (6.8)	2249	4.1-11.6 (7.1)	834	^†	2247	2.1-8.0 (4.3)
NHBF	329	3.7-10.1 (6.2)	344	3.4-11 (6.3)	329	1.2-6.6 (3.1)	344	1.4-7.5 (3.4)
MAF	401	4.1-12.0 (7.6)	552	4.5-11.8 (7.5)	401	1.9-8.4 (5.3)	550	2.2-8.0 (4.9)
NHWF	322	4.1-11.3 (7.2)	1222	3.9-11.6 (7.0)	322	1.9-7.7 (4.0)	1221	2.0-7.9 (4.7)
MAF, NHWF	723	^†	1774	^†	723	^†	1771	2.0-7.9 (4.8)

ANC, Absolute neutrophil count; MAF, female Mexican American; MAM, male Mexican American; NHBF, female non-Hispanic black; NHBM, male non-Hispanic black; NHWF, female non-Hispanic white; NHWM, male non-Hispanic white; WBC, white blood cell.

From Lim EM, Cembrowski G, Cembrowski M, et al: Race-specific WBC and neutrophil count reference intervals. J Lab Clin Hematol 32:590–597, 2010.

* Data are presented as reference intervals (2.5-97.5 percentile limits) and mean values (in parentheses).

† Could not be pooled because of variation within and between populations.

APPENDIX 18

Activities of RBC Enzymes

Enzyme	Activity at 37° C (mean ± SD)
Acetylcholinesterase	36.93 ± 3.83
Adenosine deaminase	1.11 ± 0.23
Adenylate kinase	258 ± 29.3
Aldolase	3.19 ± 0.86
Bisphosphoglyceromutase (2,3- diphosphoglyceromutase)	4.78 ± 0.65
Catalase	15.3 ± 2.4 × 10 ⁴
Enolase	5.39 ± 0.83
Epimerase	0.23 ± 0.06
Galactokinase	0.029 ± 0.004
Galactose-1-phosphate uridyltransferase	28.4 ± 6.94
Glucose phosphate isomerase	60.8 ± 11.0
Glucose-6-phosphate dehydrogenase	8.34 ± 1.59
WHO method	12.1 ± 2.09
Glutamic oxaloacetic transaminase without PLP	3.02 ± 0.67
Glutamic oxaloacetic transaminase with PLP	5.04 ± 0.90
γ-Glutamylcysteine synthetase	0.43 ± 0.04
Glutathione peroxidase	30.82 ± 4.65
Glutathione reductase without FAD	7.18 ± 1.09
Glutathione reductase with FAD	10.4 ± 1.50
Glutathione S-transferase	6.66 ± 1.81
Glyceraldehyde phosphate dehydrogenase	226 ± 41.9
Hexokinase	1.78 ± 0.38
Hypoxanthine phosphoribosyltransferase	1.72 ± 0.30
Lactate dehydrogenase	200 ± 26.5
Methemoglobin reductase	2.60 ± 0.71
Monophosphogylceromutase	37.71 ± 5.56
NADH-methemoglobin reductase	19.2 ± 3.85 (30°)
NADPH diaphorase	2.26 ± 0.16
Phosphofructokinase	11.01 ± 2.33
Phosphoglucomutase	5.50 ± 0.62
Phosphoglycerate kinase	320 ± 36.1
Phosphoglycolate phosphatase	1.23 ± 0.10
Pyrimidine 5′ nucleotidase	0.11 ± 0.03
Pyruvate kinase	15.0 ± 1.99
6-Phosphogluconate dehydrogenase	8.78 ± 0.78
Triose phosphate isomerase	2111 ± 397

FAD, Flavin adenine dinucleotide; NADH, reduced nicotinamide adenine dinucleotide; NADPH, reduced nicotinamide adenine dinucleotide phosphate; PLP, pyridoxal 5-phosphate; SD, standard deviation; WHO, World Health Organization.

From Beutler E, Blum KG: In Altman PL, Dittmer DS, editors: Human health and disease. Bethesda, Md., Federation of American Societies for Experimental Biology, 1977, p. 156; and Beutler E: Red cell metabolism: a manual of biochemical methods, 3 ^rded, Orlando, Fla., Grune & Stratton, 1984.

APPENDIX 19

Levels of Intermediate Metabolites in Normal Adult Erythrocytes

Metabolite	CONCENTRATION (mean ± 1 SD)
Metabolite	Abbreviation	nmol/g Hb	nmol/mL Red Cells	µmol/L in Whole Blood
Adenosine-5′-diphosphate	ADP	6635 ± 105	216 ± 36	—
Adenosine-5-monophosphate	AMP	62 ± 10	21.1 ± 3.4	—
Adenosine-5-triphosphate	ATP	4230 ± 290 (whites) 3530 ± 301 (blacks)	1438 ± 99 1200 ± 102	—
2,3-Diphosphoglycerate	2,3-DPG	1227 ± 1870	4171 ± 636	—
Glutathione	GSH	6570 ± 1040	2234 ± 354	—
Glutathione (oxidized)	GSSG	12.3 ± 4.5	4.2 ± 1.53	—
Glucose-6-phosphate	G6P	82 ± 22	27.8 ± 7.5	—
Fructose-6-phosphate	F6P	27 ± 5.8	9.3 ± 2.0	—
Fructose-6-diphosphate	FDP	5.6 ± 1.8	1.9 ± 0.6	—
Dihydroxyacetone phosphate	DHAP	27.6 ± 8.2	9.4 ± 2.8	—
3-Phosphoglyceric acid	3-PGA	132 ± 15.0	44.9 ± 5.1	—
2-Phosphoglyceric acid	2-PGA	21.5 ± 7.35	7.3 ± 2.5	—
Phosphoenolpyruvate	PEP	35.9 ± 6.47	12.2 ± 2.2	—
Creatine		1310 ± 310 (male) 1500 ± 250 (female)	445 ± 105 510 ± 85	—
Lactate		—	—	932 ± 211
Pyruvate		—	—	53.3 ± 21.5

SD, Standard deviation.

From Beutler E : Red cell metabolism: a manual of biochemical methods, 3rd ed. Orlando, Fla., 1984, Grune & Stratton.

APPENDIX 20

Red Cell Enzyme Activity in Premature and Term Neonates, Adults, and Adult Reticulocytes

	Premature Infants (N = 11)	Term Infants (N = 10)	Normal Adults (N = 50)	95% Adult * Reticulocytes
Hexokinase (HK)	3.6 ± 0.6 ^a	2.7 ± 0.3 ^a	1.15 ± 0.35	9.9 ^a
Glucosephosphate isomerase (PGI)	94.5 ± 15.2 ^a	88 ± 16 ^a	67.7 ± 12.0	95.7 ^c
Phosphofructokinase (PFK)	8.0 ± 1.3 ^d	7.1 ± 1.5 ^c	8.4 ± 3.4	14.1 ^d
Aldolase (ALD)	4.3 ± 0.8 ^c	5.3 ± 1.7 ^b	3.55 ± 1.07	9.5 ^a
Triosephosphate isomerase (TIM)	2961 ± 1378 ^d	1787 ± 380 ^d	2700 ± 700	3220 ^d
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)	131 ± 76 ^d	164 ± 45 ^d	134 ± 93	240 ^d
Phosphoglycerate mutase (PGK)	591 ± 114 ^b	436 ± 119 ^b	312 ± 86	438 ^d
Diphosphoglycerate mutase (DPGM)	8.3 ± 3.6 ^c	6.7 ± 1.9 ^c	5.3 ± 2.5	7.3 ^d
Enolase (ENO)	15.1 ± 3.4 ^a	14.6 ± 2.1 ^a	7.8 ± 2.2	8.1 ^d
Pyruvate kinase (PK)	19.4 ± 3.5 ^a	17.3 ± 2.5 ^a	11.4 ± 2.8	54.2 ^a
Glucose-6-phosphate dehydrogenase (G6PD)	21.3 ± 3.8 ^a	19.1 ± 4.3 ^a	10.9 ± 3.8	30.6 ^a
6-Phosphogluconate dehydrogenase (6PGD)	9.1 ± 2.1 ^d	9.9 ± 1.6 ^c	8.4 ± 1.0	14.4 ^a
Glutathion reductase (GR)	9.5 ± 1.7 ^a	11.0 ± 2.6 ^a	7.2 ± 2.2	11.3 ^c
Acetyl choline esterase (ACE)	17.5 ± 3.5 ^a	18.5 ± 4.1 ^a	36.9 ± 3.8	44.4 ^c

From Jansen G, Koenderman L, Rijksen G, et al: Characteristics of hexokinase, pyruvate kinase, and glucose-6-phosphate dehydrogenase during adult and neonatal reticulocyte maturation. Am J Hematol 20:203–215, 1985.

* Obtained after Percoll density gradient centrifugation of red cells from a patient with a hemolytic syndrome. Statistical interpretation versus normal adult group: P <.001; P <.01; P <.1; not significant.

APPENDIX 21

Red Cell Glycolytic Intermediate Metabolites in Normal Adults, Term Infants, and Premature Infants *

Metabolite	Normal Adults (10)	Term Infants (10)	Premature Infants (11)	Normals (5)
Glucose-6-phosphate	24.8 ± 9.8	45.2 ± 8.7	66.8 ± 34.8	27 ± 2.4
Fructose-6-phosphate	5.4 ± 1.0	9.9 ± 2.3	20.5 ± 8.9	11 ± 2.5
Fructose, 1,6-diphosphate	4.6 ± 1.0	3.8 ± 0.7	3.6 ± 0.8	5 ± 0.9
Dihydroxyacetone phosphate	4.9 ± 3.5	11.9 ± 5.0	18.6 ± 10.7	12 ± 3.7
Glyceraldehyde-3-phosphate	2.6 ± 0.7	1.9 ±1.6	6.5 ± 3.2	4 ± 1.5
3-Phosphoglycerate	61.6 ± 12.4	58.2 ± 14.4	47.5 ± 14.2	48 ± 16.1
2-Phosphoglycerate	4.3 ± 1.8	4.9 ± 1.6	4.4 ± 2.5	7 ± 1.7
Phosphoenolpyruvate	8.8 ± 2.6	7.6 ± 2.9	7.4 ± 3.0	12 ± 0.9
Pyruvate	73.5 ± 33.1	70.4 ± 32.3	78.4 ± 4.15	71 ± 17.7
2,3-Diphosphoglycerate	4423 ± 1907	3609 ± 800	3152 ± 2133	4000

From Oski FA: Red cell metabolism in the newborn infant: V. Glycolytic intermediates and glycolytic enzymes. Pediatrics 44:87, 1969.

* Infant samples were obtained from newborns weighing more than 2800 g whose gestational age was 39 weeks or greater. Blood was drawn within 24 hours of birth. All the newborns were clinically healthy. Adult samples were obtained from healthy normal volunteers.

APPENDIX 22

RBC Enzyme Activity * in ELBWI Infants and Premature and Normal Adults (Whole Blood and Reticulocyte Rich)

	Normal Adult (N = 28) Mean ± SD	ELBWI (N = 28) Mean ± SD	Reticulocyte Rich (N = 20) Mean ± SD	ELBWI Versus Reticulocyte Rich	ELBWI Versus Normal Adults
Hexokinase (HK)	1.05 ± 0.11	3.28 ± 0.47	3.45 ± 1.78	NS	High ( P <.0001)
Glucosephosphate isomerase (GPI)	58.4 ± 5.0	83.4 ± 6.2	65.2 ± 8.5	High ( P <.0001)	High ( P <.0001)
Phosphofructokinase (PFK)	13.3 ± 1.7	10.1 ± 1.6	15.8 ± 2.4	Low ( P <.0001)	Low ( P <.0001)
Aldolase (ALD)	3.09 ± 0.53	4.73 ± 5.13	5.12 ± 0.94	NS	High ( P <.0001)
Triosephosphate isomerase (TPI)	1760 ± 253	2215 ± 339	2077 ± 375	NS	High ( P <.0001)
Glyceraldehyde-3-phosphate dehydrogenase (GA3PD)	191 ± 34	252 ± 39	232 ± 51	NS	High ( P <.0001)
Phosphoglycerate kinase (PGK)	288 ± 31	469 ± 67	355 ± 43	High ( P <.0001)	High ( P <.0001)
Monophosphoglycerate mutase (MPGM)	23.5 ± 2.8	33.8 ± 6.9	29.9 ± 5.3	High ( P = .0412)	High ( P <.0001)
Enolase (ENL)	6.9 ± 1.0	23.9 ± 3.0	9.3 ± 1.7	High ( P <.0001)	High ( P <.0001)
Pyruvate kinase (PK)	16.3 ± 2.1	29.7 ± 4.0	27.6 ± 8.1	NS	High ( P <.0001)
Lactate dehydrogenase (LDH)	195 ± 17	248 ± 28	264 ± 34	NS	High ( P <.0001)
Glucose-6-phosphate dehydrogenase (G6PD)	7.6 ± 0.8	18.1 ± 2.2	12.8 ± 2.0	High ( P <.0001)	High ( P <.0001)
6-Phosphogluconate dehydrogenase (6PGD)	8.1 ± 1.0	8.2 ± 1.5	13.2 ± 1.8	Low ( P <.0001)	NS
Glutathione reductase (GR)	7.1 ± 1.2	9.2 ± 1.8	7.8 ± 1.8	High ( P = .0090)	High ( P <.0001)
GR + flavin adenine dinucleotide (FAD)	9.9 ± 1.6 ^b	10.9 ± 1.7 ^b	10.4 ± 1.7	NS	NS
Glutathione peroxidase (GSH-Px)	31.8 ± 5.7	22.5 ± 4.0	41.3 ± 7.4	Low ( P <.0001)	Low ( P <.0001)
Adenylate kinase (AK)	252 ± 35	132 ± 18	253 ± 26	Low ( P <.0001)	Low ( P <.0001)
Adenosine deaminase (ADA)	1.25 ± 0.33	0.96 ± 0.30	1.60 ± 0.60	Low ( P <.0001)	Low ( P = .0010)
Acetylcholinesterase (AchE)	31.7 ± 4.7	16.7 ± 3.3	37.6 ± 10.1	Low ( P <.0001)	Low ( P <.0001)
Pyrimidine 5′-nucleotidase	9.1 ± 1.4	17.0 ± 4.0	19.1 ± 4.5	ns	High ( P <.0001)
NADH methemoglobin reductase (MetHbR)	14.9 ± 1.8 ^b	11.5 ± 2.1 ^a	14.1 ± 2.3 ^a	Low ( P = .0087)	Low ( P <.0001)
Catalase (CAT)	13.6 ± 2.1 ^b	11.8 ± 2.7 ^a	15.6 ± 3.2 ^a	Low ( P = .0048)	Low ( P = .0459)
Reduced glutathione (GSH) (mg/dL RBC)	67.8 ± 9.3	76.9 ± 13.8	69.4 ± 12.0	NS	High ( P = .0059)

ELBWI, Extremely low-birth-weight infants (birth weight <1000 g); Hb, hemoglobin; NS, not significant; RBC, red blood cell; SD, standard deviation.

From Miyazono Y, Hirono A, Miyamoto Y, et al: Erythrocyte enzyme activities in cord blood of extremely low-birth-weight infants. Am J Hematol 62:88–92, 1999.

* All enzyme activities are represented as units/g Hb, except for CAT (10 ⁴units/g Hb) and P5N (µmol Pi liberated/hr/g Hb). N = 12; N = 20.

APPENDIX 23

G6PD Reference Values for G6PD-Normal and –Deficient Males and Females with Probable Genotypes

Category (U/g Hb)	No.	Median Value	IQR	Min-Max	G6PD Phenotype	Probable Genotype * ^†
Male (N = 1502)
<7.0	243 (16.2%)	0.28	0.21-0.39	0.01-6.20	Deficient	Deficient hemizygote
≥9.0	1256 (83.8%)	18.76	17.27-21.68	9.00-34.66	Normal	Normal hemizygote
Female (N = 1298)
<7.0	81 (6.2%)	4.84	0.45-6.16	0.06-6.96	Deficient	Deficient homozygote
7.0-9.4	64 (4.9%)	8.61	7.67-9.00	7.04-9.46	Intermediate	Heterozygote
≥9.5	1153 (88.8%)	18.36	16.21-20.63	9.54-35.50	Normal	Normal homozygote

* In females, because of nonrandom X chromosome inactivation among heterozygotes, accurate genotype categories may be difficult to determine based on phenotype. The term “probable” genotype is therefore used.

† Male genotype based on phenotype may be assumed with reasonable accuracy.

APPENDIX 24

Hemoglobin A Content in Male and Female Newborns According to Birth Weight

Birth Weight (g)	Males Hb A (%)	Females Hb A (%)
<1501	7.1 ± 1.3 (9)	13.2 ± 11.3 (14)
1501-2000	12.2 ± 8.2 (36)	13.1 ± 10.4 (44)
2001-2500	12.6 ± 5.2 (139)	14.9 ± 6.5 (206) *
2501-3000	15.8 ± 6.4 (635)	17.8 ± 6.2 (776) *
3001-3500	18.2 ± 6.5 (1289)	19.9 ± 6.8 (1204) *
3501-4000	20.0 ± 6.6 (803)	20.9 ± 7.2 (590) *
4001-4500	19.7 ± 6.3 (200)	22.6 ± 8.6 (94) *
>4500	21.7 ± 7.0 (45)	20.9 ± 7.2 (20)

HbA, Hemoglobin A.

From Galacteros F, Guilloud-Bataille M, Feingold J: Sex, gestational age, and weight dependency of adult hemoglobin concentration in normal newborns. Blood 78:1121, 1991.

* P <.05 between male and female newborns.

APPENDIX 26

Percentage of Hemoglobins F and A2 in the Newborn and Adult *

	% Hb F (Gα : Aα ratio)	% Hb A2
Newborn	60-90 (3 : 1)	<1.0
Adult	<1.0 (2 : 3)	1.6-3.5

Hb, Hemoglobin.

From Altman PL, Dittmer DS, editors: Human health and disease. Bethesda, MD, 1977, Federation of American Societies for Experimental Biology, p. 159.

* The α chains of fetal hemoglobin contain either a glycyl residue or an alanyl residue at position 136. The Gα : Aα ratio in the newborn undergoes a considerable change between the third and fourth months of life, at which time it approximates that of the Hb F of adults.

APPENDIX 27

Methemoglobin Levels in Normal Children *

	METHEMOGLOBIN (g/dL)					METHEMOGLOBIN AS PERCENTAGE OF TOTAL HEMOGLOBIN
	No. Cases	No. Det.	Mean	Range	Standard Dev.	No. Cases	No. Det.	Mean	Range	Standard Dev.
Prematures (birth-7 days)	29	34	0.43	(0.02-0.83)	±0.07	24	28	2.3	(0.08-4.4)	±1.26
Prematures (7-72 days)	21	29	0.31	(0.02-0.78)	±0.19	18	23	2.2	(0.02-4.7)	±1.07
Prematures (total)	50	63	0.38	(0.02-0.83)	±0.10	42	51	2.2	(0.08-4.7)	±1.10
Cook County Hospital, prematures (1-14 days)	8	8	0.52	(0.18-0.83)	±0.08	—	—	—	—	—
Newborns (1-10 days)	39	39	0.22	(0.00-0.58)	±0.17	25	30	1.5	(0.00-2.8)	±0.81
Infants (1 month-1 year)	8	8	0.14	(0.02-0.29)	±0.09	8	8	1.2	(0.17-2.4)	±0.78
Children (1-14 years)	35	35	0.11	(0.00-0.33)	±0.09	35	35	0.79	(0.00-2.4)	±0.62
Adults (14-78 years)	30	30	0.11	(0.00-0.28)	±0.09	27	27	0.82	(0.00-1.9)	±0.63

From Kravitz H, Elegant LD, Kaiser E, et al: Methemoglobin values in premature and mature infants and children. Am J Dis Child 91:2, 1956.

* The premature and full-term infants were free of known disease. None had respiratory distress or cyanosis. Analysis of milk and water ingested by these infants revealed a nitrate level less than 0.027 ppm. The premature infants routinely received vitamin C orally each day from the seventh day of life.

APPENDIX 28

Hemoglobin, Serum Transferrin Saturation, Serum Ferritin * , and Red Cell Indices in Adolescents (15-16 Years of Age) ^†

		PERCENTILE
	Mean ± SD	10th	25th	50th	75th	90th
Boys
Hb (g/dL)	14.7 ± 0.831	135	14.2	14.7	15.2	15.7
Transferrin saturation (%)	32.7 ± 10.25	21.2	25.7	31.7	38.6	46.3
Serum ferritin (µg/L)	26.4 ± 17.71	13.2	22	29	40.8	52
MCV (fL)	88.4 ± 3.87	83.5	85.5	88.1	90.9	93.2
RDW (%)	13.0 ± 0.95	12.3	12.5	12.9	13.2	13.5
MCH (pg)	29.4 ± 1.43	27.7	28.6	29.2	30.2	31.0
Girls
Hb (g/dL)	13.4 ± 0.763	12.3	12.9	13.4	13.9	14.3
Transferrin saturation (%)	29.9 ± 10. 7	17.2	22.8	29	35.2	43.4
Serum ferritin (µg/L)	18.2 ± 1.98	7.5	12	18.2	28.5	41.5
MCV (fL)	90.1 ± 3.86	84.8	87.6	90.3	92.8	94.8
RDW (%)	12.8 ± 0.71	12.1	12.4	12.7	13.2	13.6
MCH (pg)	29.6 ± 1.43	27.8	28.8	29.5	30.6	31.3

Hb, Hemoglobin; MCH, mean corpuscular hemoglobin; MCV, mean corpuscular volume; RDW, red blood cell volume distribution width; SD, standard deviation.

From Hallberg L, Hulten L, Lindstedt G, et al: Prevalence of iron deficiency in Swedish adolescents. Pediatr Res 34:680–687, 1993.

* Data for serum ferritin are presented as geometric means ± antilog of logarithmic values.

† Data collected in 197 to 207 boys and 215 to 220 girls, from ages 15 to 16.

APPENDIX 29

Hemoglobin and Iron Status Indicators According to Age

	2 m			6 m			9 m
	Mean ± SD	5-95 Percentiles	N	Mean ± SD	5-95 Percentiles	N	Mean ± SD	5-95 Percentiles	N
Hemoglobin
(g/L)	115 ± 8	101-130	68	116 ± 9	105-130	57	117 ± 7	103-129	94
(mmol/L)	7.1 ± 0.5	6.3-8.1		7.2 ± 0.6	6.5-8.1		7.3 ± 0.5	6.4-8
Serum ferritin * (µg/L)	301	134-675	63	59	23-140	54	37	17-78	84
Erythrocyte protoporphyrin (nmol/l erythrocytes)	679 ± 185 ^†	479-1148	65	475 ± 109	304-659	48	556 ± 133 ^†	369-763	83
Serum transferrin (µmol/L)	26 ± 4 ^†	20-33	65	31 ± 4	24-38	57	35 ± 7 ^†	26-47	90
Serum iron (µmol/L)	13.1 ± 3.5 ^†	7.5-19.1	59	9.9 ± 3.1	6.3-17.3	46	8.8 ± 3.4	3.9-14.9	80
Transferrin saturation (%)	26 ± 8 ^†	15-40	59	16 ± 6	10-29	46	13 ± 6	5-25	77
Mean red cell volume (fl)	86 ± 5 ^†	78-94	64	76 ± 4	69-82	49	76 ± 6	68-85	89

m , Months; SD , standard deviation.

From Michaelsen KF, Milman N, Samuelson G: A longitudinal-study of iron status in healthy Danish infants—effects of early iron status, growth velocity and dietary factors. Acta Paediatr 84:1035–44, 1995.

* Geometric mean values.

† Significantly different ( P <.001) from values at 6 months (Student’s paired t -test).

APPENDIX 30

Serum Ferritin, Iron, Total Iron-Binding Capacity, and Transferrin Saturation in Children and Adolescents

Age		Male Subjects				Female Subjects
Ferritin	ng/mL		µg/L		ng/mL		µg/L
1-30 d *	6-400		6-400		6-515		6-515
1-6 m *	6-410		6-410		6-340		6-340
7-12 m *	6-80		6-80		6-45		6-45
1-5 y ^†^‡	6-24		6-24		6-24		6-24
6-9 y ^†^‡	10-55		10-55		10-55		10-55
10-14 y ^‡	23-70		23-70		6-40		6-40
14-19 y ^‡	23-70		23-70		6-40		6-40
Iron	µg/dL		µmol/L		µg/dL		µmol/L
1-5 y ^†^‡	22-136		4-25		22-136		4-25
6-9 y ^†^‡	39-136		7-25		39-136		7-25
10-14 y ^‡	28-134		5-24		45-145		8-26
14-19 y ^‡	34-162		6-29		28-184		5-33
Iron-Binding Capacity
1-5 y ^†^‡	268-441		48-79		268-441		48-79
6-9 y ^†^‡	240-508		43-91		240-508		43-91
10-14 y ^‡	302-508		54-91		318-575		57-103
14-19 y ^‡	290-570		52-102		302-564		52-101
Transferrin Saturation
1-5 y ^†^‡		0.07-0.44				0.07-0.44
6-9 y ^†^‡		0.17-0.42				0.17-0.42
10-14 y ^‡		0.11-0.36				0.02-0.40
14-19 y ^‡		0.06-0.33				0.06-0.33
Transferrin				U/L (Males and Females)
0-5 d ^§				1.43-4.46
1-3 y ^§				2.18-3.47
4-6 y ^§				2.08-3.78
7-9 y ^§				2.25-3.61
10-13 y ^§				2.24-4.42
14-19 y ^§				2.33-4.44

d, Days; m, months; y, years.

* Soldin SJ, Morales A, Albalos F, et al: Pediatric reference ranges on the Abbott IMx for FSH, LH, prolactin, TSH, T _4,T ₃, free T3, T-uptake, IgE, and ferritin. Clin Biochem 28:603, 1995. Study based on hospitalized patients; values represent 2.5th and 97.5th percentiles.

† No significant differences between males and females; range derived from combined data.

‡ Lockitch, G; Halstead, AC; Wadsworth, et al: Age- and sex-specific pediatric reference intervals and correlations for zinc, copper, selenium, iron, vitamins A and E, and related proteins. Clin Chem 34:1625, 1988. Study based on healthy children; values represent the 0.025 and 0.975 fractiles. Transferrin saturation calculated from iron (µmol/L)/total iron binding capacity. Note that the lower reference limits for serum iron and transferrin saturation in this study are below the limits used to define acceptable levels for these two analytes (O’Neal RM, Johnson OC, Schaefer AE: Guidelines for classification and interpretation of group blood and urine data collected as part of the National Nutrition Survey. Pediatr Res 4:103, 1970.)

§ Lockitch, G; Halstead, AC; Quigley G, et al: Age- and sex-specific pediatric reference intervals: study design and methods illustrated by measurement of serum proteins with the Behring LN nephelometer. Clin Chem 34:1618, 1988. Results are 2.5th and 97.5 percentiles.

APPENDIX 31

Reference Ranges for Erythrocyte Zinc Protoporphyrin in Children and Adolescents

Age Groups	Blood ZPP Reference Ranges 2.5th–97.5th Percentiles (µg/dL) *		Blood ZPP Reference Ranges 2.5th–97.5th Percentiles (µmol/mol heme)		N
Age Groups	Males	Females	Males	Females	Males	Females
0-12 months	8.5-34.5	9-40	15.6-63.5	16.6-73.6	145	203
13-24 months	10-34	11-32	18.4-62.6	20.2-58.9	605	725
2-5 years	5-35	10-31	9.2-64.4	18.4-57.0	1926	1822
6-9 years	6-31	9-30	11.0-57.0	16.6-55.2	522	408
10-17 years	5.5-31.5	5-33.5	10.1-58.0	9.2-61.6	61	61
Total					3259	3219

ZPP, Zinc protoporphyrin.

From Soldin OP, Miller M, Soldin SJ: Pediatric reference ranges for zinc protoporphyrin. Clin Biochem 36:21–25, 2003.

* Multiply µg/dL by 1.83 to convert to µmol/mol heme.

APPENDIX 32

Reference Ranges for Serum Transferrin Receptor in Children and Adolescents *

Age Groups	2.5% Reference Limit mg/L	97.5% Reference Limit mg/L
6 months to 4 years	1.5 (1.4-1.5)	3.3 (3.1-3.4)
4 to 10 years	1.3 (1.3-1.4)	3.0 (2.9-3.2)
10 to 16 years	1.2 (1.1-1.2)	2.7 (2.7-2.8)
>16 years	0.9 (0.9-1.0)	2.3 (2.2-2.4)

From Suominen, P, Virtanen A, Lehtonen-Veronmaa M, et al: Regression-based reference limits for serum transferrin receptor in children 6 months to 16 years of age. Clin Chem 47:935–937, 2001.

* The 95% confidence interval for each limit is given in parentheses.

APPENDIX 33

Percentile Distributions for Plasma Folate, RBC folate, Vitamins B ₆and B ₁₂, Holotranscobalamin, and Total Homocysteine of Healthy European Adolescents 12.5 to 17.49 Years of Age

		PF(nmol/L)	RBC Folate nmol/L	PLP (nmol/L)	Cbl (pmol/L)	Holo-TC (pmol/L)	tHcy (µmol/L)
	N	1049	1040	974	1051	1017	1050
	Mean	18.7	784.2	63.2	349.4	63.4	7.4
	SD	10.5	340.9	51.9	143.5	33.7	3.8
Percentile	2.5	6.4	315.5	16.6	154.4	27.4	3.7
	5	7.6	378.8	20.0	170.8	30.9	3.9
	10	9.1	428.9	25.0	193.0	35.2	4.4
	25	11.8	553.0	34.7	240.0	44.4	5.4
	50	16.0	721.9	49.1	319.0	57.8	6.7
	75	22.1	942.3	75.7	434.9	73.0	8.4
	90	31.7	1214.0	113.9	557.0	91.7	10.5
	95	39.0	1389.3	149.7	638.0	107.1	12.3
	97.5	46.4	1582.6	187.3	696.0	127.1	15.7

Cbl, Cobalamin; Holo-TC, holotranscobalamin; PF, plasma folate; PLP, pyridoxal 5-phosphate; RBC, red blood cell; SD, standard deviation.

APPENDIX 34

Plasma Levels of Vitamin E (α-Tocopherol) in Children and Adolescents

Age	MALES AND FEMALES
Age	µmol/L	µg/mL
Premature *	1-8	0.5-3.5
Full Term	2-8	0.5-3.5
2-5 m *	5-14	2.0-6.0
6-24 m *	8-19	3.5-80
2-12 y *	13-21	5.5-9.0
1-6 y ^†	7-21	3.0-9.0
7-12 y ^†	10-21	4.0-9.0
13-19 y ^†	13-24	6.0-10.1

* Meites S, editor: Pediatric clinical chamistry, 3rd ed. Washington, DC, 1989, AACC Press, p. 295–296.

† Lockitch G, Halstead AC, Wadsworth L, et al: Age- and sex-specific pediatric reference intervals for zinc, copper, selenium, iron, vitamins A and E and related proteins. Clin Chem 34:1625, 1988. Study based on healthy children; values represent the 0.025th and 0.975th fractiles.

APPENDIX 35

Serum Erythropoietin Levels During the First Year of Life *

Days After Birth	Erythropoietin (mU/mL)	Hemoglobin (g/dL)	RBC (×10 ⁶/µL)
0-6	33.0 ± 31.4 (11)	15.6 ± 2.2 (11)	4.51 ± 0.74 (11)
7-50	11.7 ± 3.6 (7)	12.8 ± 1.1 (5)	3.92 ± 0.35 (5)
51-100	21.1 ± 5.5 (13)	11.4 ± 1.0 (10)	4.09 ± 0.51 (10)
101-150	15.1 ± 3.9 (5)	11.2 ± 1.1 (3)	4.21 ± 0.31 (3)
151-200	17.8 ± 6.3 (6)	—	—
>201	23.1 ± 9.7 (10)	11.8 ± 0.8 (9)	4.57 ± 0.25 (9)

RBC, Red blood cell.

From Yamashita H, Kukita J, Ohga S: Serum erythropoietin levels in term and preterm infants during the first year of life. Am J Pediatr Hematol Oncol 16:213, 1994.

* Values are expressed as mean ± standard deviation; number of specimens is within parentheses.

APPENDIX 36

Plasma Erythropoietin Reference Ranges in Children and Adolescents *

Age (y)	MALE SUBJECTS		FEMALE SUBJECTS
Age (y)	2.5%	97.5%	2.5%	97.5%)
1-3	1.7	17.9	2.1	15.9
4-6	3.5	21.9	2.9	8.5
7-9	1.0	13.5	2.1	8.2
10-12	1.0	14.0	1.1	9.1
13-15	2.2	14.4	3.8	20.5
16-18	1.5	15.2	2.0	14.2

y, Years.

From Krafte-Jacobs B, Williams J,. Soldin SJ, Plasma erythropoietin reference ranges in children. J Pediatr 126:601, 1995.

* Data obtained from a total of 1122 hospitalized and outpatient children ages 1 to 18 years. Levels found in anemic patients cannot be compared with normal values. In fact, as long as the erythropoietin-generating apparatus in the kidney is efficient, serum levels increase exponentially as the hematocrit concentration decreases. Serum erythropoietin must therefore be evaluated in relation to the degree of anemia, and every single laboratory should determine the exponential regression of serum erythropoietin versus hematocrit (or hemoglobin) in a home-made reference population of anemic subjects and define the 95% confidence limits. The patients gathered to calculate a reference regression equation should have an anemia with a single simple mechanism and no evidence of either renal failure or excessive cytokine production (i.e., normal values for C-reactive protein and α ₂-globulins). Patients with chronic iron-deficiency anemia caused by nonneoplastic and noninflammatory chronic blood loss have the advantages of being easily found, unequivocably defined, and homogeneous. They could become the universal reference population, although patients with hemolytic anemia or thalassemia intermedia also may be studied as reference subjects. Serum erythropoietin levels are also much higher for hemoglobin concentration in hypoplastic than in hyperplastic states. Thus for the same hemoglobin concentration, serum erythropoietin levels are lower in thalassemia intermedia than in Diamond-Blackfan anemia.