The mean corpuscular volume (MCV), red blood cell distribution width (RDW), and the patient's history can exclude some of these etiologies. These cookies track visitors across websites and collect information to provide customized ads. When HbF is higher than 2% of the total hemoglobin, then this is considered abnormal. Available from: National Heart, Lung, and Blood Institute [Internet]. dog hair dander allergen. The hemoglobin electrophoresis with beta thalassemia trait usually has reduced or absent HbA, elevated levels of HbA2, and increased HbF.2 However, a normal concentration of HbA2 does not rule out beta thalassemia trait, especially if there was coexistent iron deficiency, which can lower HbA2 levels into the normal range. If thalassemia is still suspected, a hemoglobin electrophoresis may help diagnose the condition. If the synthesis of beta chains is less severely reduced, the person has beta thalassemia intermedia. Test Code: 121690 Specimen Type: Blood Description: Hemoglobin Electrophoresis Blood Test (Labcorp) The Hemoglobin Electrophoresis test is used to detect and quantify both normal and abnormal forms of Hemoglobin . Hemoglobin electrophoresis alone is not adequate for diagnosing thalassemia. In our laboratory, range of HbA2 value with normal RBC parameter was 2.0-3.3%. Lab test ranges: definitions. Hemoglobin Electrophoresis; [cited 2021 Aug 11]; [about 3 screens]. Pediatrics. Interpretation of hemoglobin electrophoresis results should be placed in the clinical context, including the family history and results of serum iron studies, red cell morphology, hemoglobin, hematocrit, and red cell indices (eg, mean corpuscular volume). Transfusions will occasionally be necessary depending upon the severity of the clinical condition. The tests measure the levels of certain enzymes and proteins in your blood. 9500 Euclid Avenue, Cleveland, Ohio 44195 |, Important Updates + Notice of Vendor Data Event, (https://www.hematology.org/education/patients/blood-basics). Previous investigators have compiled information on normal laboratory reference ranges in pregnancy [30,40,43,76]. A human body contains various types of hemoglobin in their blood. American Society of Hematology [Internet]. Normal value ranges are: Total protein: 6.4 to 8.3 grams per deciliter (g/dL) or 64 to 83 grams per . Family physicians need to know how to diagnose thalassemias, how to distinguish them from other causes of a microcytic anemia, and the treatment options for severe forms of thalassemia. Minor adult hemoglobin (HbA2) is made of two - and two -globin chains (2 2). There are several different types of hemoglobin. If levels of HgbA or HgbF are too high or too low, it can indicate certain types of anemia. Sometimes, the gene controlling your hemoglobin changes or mutates, turning healthy red blood cells into damaged cells. To diagnose blood disorders: Your doctor may have you do a hemoglobin electrophoresis test if youre showing symptoms of anemia. 2 nd trimester of pregnancy: 9.7 to 14.8 g/dl or 97 to 148 g/L. We will review it in the next few hours. Depending on the type of hemoglobin affected, thalassemia can cause mild, moderate, or severe anemia. Surgery is usually delayed until at least four years of age because of the spleen's role in clearing bacteria and preventing sepsis. Hemoglobin (HEE-muh-glow-bin) is the protein inside red blood cells that carries oxygen. Most persons with thalassemia trait are found incidentally when their complete blood count shows a mild microcytic anemia. (III-A) 3. They may use finger stick devices or heel sticks to get blood from your child or newborn baby. Well go over normal hemoglobin ranges for adults, children, and, A serum hemoglobin test measures the amount of free-floating hemoglobin in your blood serum. Cleveland (OH): Cleveland Clinic; c2020. The most common types of normal hemoglobin are: Hemoglobin A. 2005-2023 Healthline Media a Red Ventures Company. Risk factors include: A health care professional will take a blood sample from a vein in your arm, using a small needle. Transfusion dependence is uncommon. This causes the different types of hemoglobin to separate into different bands. Many of these conditions can be treated if found early. You should be able to increase hemoglobin in pregnancy by taking iron supplements. Available from: Salinas Cisneros G, Thein SL. An electric current is passed through the blood sample, which helps to segregate different hemoglobin into separate bands. [3] Citrate agar electrophoresis at pH 6.0-6.2 provides better resolution for different hemoglobin variants. Your blood sample is finally compared to a normal hempglobin sample to give test results. The hemoglobin electrophoresis testsfor abnormal forms of hemoglobin (hemoglobinopathy). There are two options for how the blood sample is tested: Hemoglobin electrophoresisThis is a test that looks at the different types of hemoglobin in your blood. Available from: Lab Tests Online [Internet]. Hemoglobin types include hemoglobin A 1 (HbA 1 ),. These cookies ensure basic functionalities and security features of the website, anonymously. Hemoglobin electrophoresis is typically done when a person has signs or symptoms of hemolytic anemia, a family history of a hemoglobinopathy, an abnormal complete blood count (CBC) test, or a positive neonatal screening test. This leads to the decrease of RBC concentration in blood, causing hemoglobin levels to drop. This condition, known as phlebitis, can be treated with a warm compress several times a day. Your doctor may use it to diagnose or monitor hemolytic, Hemoglobin is a protein in your red blood cells that carries oxygen to the rest of your body. Thalassemia Genetic Testing by PCR-Based Methods PCR-based methods, including gap-PCR and PCR-RDB, were carried out at 9 study centers according to the manufacturer's protocol (Yanengbio, Yishengtang, Kaipu Bioscience). Healthcare providers place dissolved red blood cells from the sample on a cellulose strip. Anemia was defined as a hemoglobin level less than 11.0 g/dl in the first and third trimester or less than 10.5 g/dl in the second trimester of pregnancy. You may also want testing if you are at risk for having a child with sickle cell disease or another inherited hemoglobin disorder. Hemoglobin levels that are too high or too low may mean: Your results may also show whether a specific disorder is mild, moderate, or severe. The U.S. Food and Drug Administration recently approved oral deferasirox (Exjade) as an alternative treatment.17 Adverse effects of deferasirox were transient and gastrointestinal in nature, and no cases of agranulocytosis were reported. Yashoda Hospitals. The distinguishing finding in beta thalassemia is a hemoglobin electrophoresis with the finding of elevated Hgb A2 and F. Both will be increased in beta thalassemia trait without iron deficiency, and will be normal or decreased in alpha thalassemia and isolated iron deficiency anemia. This lab test measures the types of protein in the fluid (serum) part of a blood sample. These include: If you've just had a baby, your newborn will be tested as part of a newborn screening. A person viewing it online may make one printout of the material and may use that printout only for his or her personal, non-commercial reference. Centers for Disease Control and Prevention. Symptoms range from mild to severe. Available from: Lab Tests Online [Internet]. Alpha thalassemia intermedia, or HbH disease, causes hemolysis and severe anemia. Approximately 5 percent of the world's population has a globin variant, but only 1.7 percent has alpha or beta thalassemia trait.2 Thalassemia affects men and women equally and occurs in approximately 4.4 of every 10,000 live births. Hemoglobin is the protein inside red blood cells responsible for transporting oxygen to your tissues and organs. What is a hemoglobin electrophoresis test? Jaundice; [updated 2019 Oct 30; cited 2020 Jan 10]; [about 2 screens]. There are usually two genes, but if there is only one gene for HgS and the other is HgA then the person is a carrier, but if there are two of the HgC or HgS genes or one is HgS and the other is HgC then the person has the actual condition. dystrophin test Hemoglobin electrophoresis is used to diagnose beta thalassemia. He or she can help you understand the disorder and your risk of passing it along to your child. Erythrocyte sedimentation rate (ESR) rises significantly during pregnancy, levels being dependent on gestational age and hemoglobin concentration. Copyright 2009 by the American Academy of Family Physicians. hemoglobin < 27 pg) in the presence of a normal hemoglobin electrophoresis or high performance liquid chromatography the patient should be investigated with a brilliant cresyl blue stained blood smear to identify H bodies. St. Louis, Mo: Elsevier; 2019. Hemoglobin Electrophoresis. Since the test results are useful to rule out sickle cell and other blood disorders, it is also called Hemoglobin evaluation test or Sickle cell screen. Persons with beta thalassemia major are almost never symptomatic at birth because of the presence of HbF, but symptoms begin to develop by six months of age. Fetal hemoglobin-containing cells have the same mean corpuscular hemoglobin as cells without fetal hemoglobin: a reciprocal relationship between gamma- and beta-globin gene expression in normal subjects and in those with high fetal hemoglobin production. Most deaths are caused by the cardiac complications of iron overload.11. Hemoglobin A1 (96% to 98% of total hemoglobin), Hemoglobin A2 (1.5% to 3.5% of total hemoglobin), Hemoglobin F (0.5% to 1% of total hemoglobin). Yashoda Hospitals is committed to providing world-class treatment for patients from across the globe. If you or your child was diagnosed with a sickle cell disease or other hemoglobin disorder, talk to your provider about treatment options. Eric B Staros, MD is a member of the following medical societies: American Medical Association, American Society for Clinical Pathology, Association for Molecular Pathology, College of American PathologistsDisclosure: Nothing to disclose. Here is the normal range of haemoglobin in adults. Too high or too low hemoglobin levels they can therefore mean: Heres the typical test procedure: Isoelectric focusing (IEF) is another technique for identifying abnormal hemoglobin. Values above the norm of hemoglobin A2 (over 3,5% of the total) may indicate the presence of beta thalassemia or Mediterranean anemia. This patient probably:-is normal-has sickle cell anemia . Remember to take them regularly as levels can go back down once you stop taking them. Too high or too low hemoglobin levels they can therefore mean: The results can also show whether a specific disorder is mild, moderate, or severe. Serum ferritin is the best test to screen for iron deficiency anemia.10 In the absence of inflammation, a normal ferritin level generally excludes iron deficiency. Bishnu Prasad Devkota, MD, MHI, FRCS(Edin), FRCS(Glasg), FACP, FAMIA is a member of the following medical societies: American College of Physicians, American Medical Informatics Association, Healthcare Information and Management Systems Society, Royal College of Physicians and Surgeons of Glasgow, Royal College of Surgeons of EdinburghDisclosure: Nothing to disclose. There is very little risk to having a blood test. Anemia of chronic disease is most often a normocytic normochromic mild anemia. dismorphic red blood cells. 80. Normal Results In adults, these hemoglobin molecules make up the following percentages of total hemoglobin: Hb A: 95% to 98% Hb A2: 2% to 3% Hb F: 0.8% to 2% Hb S: 0% Hb C: 0% In infants and children, these hemoglobin molecules make up the following percentages of total hemoglobin: Hb F (newborn): 50% to 80% Hb F (6 months): 8% Hemoglobin A 2 levels have special application to the diagnosis of -thalassemia trait, which may be present even though peripheral blood smear is normal. Hemoglobin electrophoresis: Overview; [updated 2020 Jan 10; cited 2020 Jan 10]; [about 2 screens]. The problem can be so severe that it can even interrupt the regular progression of the gestation, causing side effects such as: As your hemoglobin levels keep falling, the symptoms will get worse and worse. Front Physiol [Internet] 2020 May 20 [cited 2021 Aug 11];11:435. The hemoglobin electrophoresis, on the other hand, offers a complete and more comprehensive breakdown of the hemoglobin in the test results. Beta thalassemia intermedia is less severe than beta thalassemia major and may require episodic blood transfusions. All material on this website is protected by copyright, Copyright 1994-2023 by WebMD LLC. Clinical and laboratory haematology. Arlington (VA): March of Dimes; c2020. Healthline has strict sourcing guidelines and relies on peer-reviewed studies, academic research institutions, and medical associations. . Test Overview. Hemoglobin electrophoresis is used as a screening test to identify normal and abnormal hemoglobins and assess their quantity. National Heart, Lung and Blood Institute. [3], Capillary electrophoresis (CE) is comparable to CA electrophoresis for reliable measurement of Hb fractions. The adequacy of the oxygen transport system depends on the affinity of hemoglobin for oxygen In adults, HbA is the major hemoglobin (97%), composed of 2 subunits and 2 subunits () with minor amount of HbA 2 (approximately 1.5 - 3.5%; 2 2) and HbF (approximately 1%; 2 2) ; Two -globin genes are located on each chromosome 16, resulting in 4 -gene loci (/) Hb Electrophoresis analysis. A patient has target cells and a positive tube solubility test. It is difficult to determine whether they are carriers of silent mutations or high normal HbA2 without genetic test. This leads to the decrease of RBC concentration in blood, causinghemoglobin levels to drop. Alpha thalassemia major with Hb Bart's causes nonimmune hydrops fetalis in utero, which is almost always fatal. This is different in babies and adults: Hemoglobin is mostly made up of hemoglobin F in fetuses. Comparison of capillary electrophoresis with cellulose acetate electrophoresis for the screening of hemoglobinopathies. ReadDr. Amos' full bio, the book about him "Lessons in Survival: All About Amos," and afictionalized account of his father's lifein the novel, "Through Walter's Lens.". Available from: Merck Manual Consumer Version [Internet]. This is the most common type of hemoglobin found normally in adults. direct donation compatibility test. No HPCL and Hemoglobin electrophoresis are not the same test. (Hb H is a tetramer of beta globin chains.) You may need testing if you have symptoms of a hemoglobin disorder. (: Hb Electrophoresis Test) Hb . Understanding how a snapshot fits into your big picture may help. Specimen: 5 mL blood in any anticoagulant. Beta thalassemia major causes hemolytic anemia, poor growth, and skeletal abnormalities during infancy. Available from: UW Health [Internet]. To test a newborn, a health care provider will clean your baby's heel with alcohol and poke the heel with a small needle. Hemoglobin (Hb) Electrophoresis by HPLC test measures and identifies different types of hemoglobin in the blood. High hemoglobin usually occurs when there is an underlying issue present. Necessary cookies are absolutely essential for the website to function properly. But now, promising new therapies have become available. To use the sharing features on this page, please enable JavaScript. 4. Chui DHK SM. All rights reserved. Normal Hgb levels also depend on the age of a person. Estimating hemoglobin levels is a common practice after you get pregnant. It also looks for abnormal types of hemoglobin. It's most often used to help diagnose anemia, sickle cell disease, and other hemoglobin disorders. Hemoglobin electrophoresis is a test that measures the different types of hemoglobin in the blood. Ongoing bleeding could be a problem if you have a bleeding disorder or are taking blood-thinning medication, such as warfarin (Coumadin) or aspirin (Bufferin). The cookie is used to store the user consent for the cookies in the category "Other. In adults, these hemoglobin molecules make up the following percentages of total hemoglobin: In infants and children, these hemoglobin molecules make up the following percentages of total hemoglobin: Dr. Amos Grunebaum, MD, FACOG is a Professor of Obstetrics and Gynecology, and among the world's leading authorities on fertility and pregnancy. Newborn screening is a group of tests given to most American babies shortly after birth. The cookie is used to store the user consent for the cookies in the category "Performance". During gestation, the body needs more oxygen, while hemoglobin is responsible for carrying oxygen and the concentration of it affects the oxygen-carrying capacity. A hemoglobin electrophoresis test is a blood test done to check the different types of hemoglobin in the blood. The normal range is just a guide. Workup relies primarily on laboratory evaluation, hemoglobin electrophoresis, and genetic testing (alpha thalassemia mutations panel). Various seeds such as flax seeds and sunflower seeds should also be a part of your diet as they are rich in folate too. Persons with thalassemia trait have a normal life expectancy. Using these references, as well as publications by other researchers in which normal values were determined across pregnancy for a number of analytes, we compiled a table of the most common, and some not so common, laboratory test . Until recently, treatment options for sickle cell disease were limited. This cookie is set by GDPR Cookie Consent plugin. Healthline Media does not provide medical advice, diagnosis, or treatment. Heavy menstrual flows that you've been experiencing before becoming pregnant, Eating a diet low in iron for some time before getting pregnant, Your body being incapable of absorbing iron normally and thus causing your hemoglobin in pregnancy to be extremely low, Coldness of the peripheral parts of the body, such as feet and hands, Respiratory difficulties, such as shortness of breath, that occur even while resting, Foods rich in iron such as leafy vegetables, dry fruits such asalmonds and raisin, sesame seeds, fruits like kiwi, peaches apples, guava, and grapefruit, Foods rich in vitamin C such as kiwi, lime, raspberries, and oranges, as well as green leafy vegetables, tomatoes, broccoli, and peppers. Apr 1987;69(4):1109-1113: Witkowska HE, Bitsch F, Shackleton CH. Alpha thalassemia major with hemoglobin Bart's usually results in fatal hydrops fetalis. A high hemoglobin F (HbF) value - in patients aged two years and over - should suggest the possible diagnosis of beta thalassemia or persistence of fetal hemoglobin (HbF). Clin Chem. These cookies will be stored in your browser only with your consent. When not pregnant: 12 to 15.8 g/dl or 120 to 158 g/L. Alpha thalassemia intermedia with significant hemoglobin H (hemoglobin H disease), Moderate to severe hemolytic anemia, modest degree of ineffective erythropoiesis, splenomegaly, variable bone changes, Alpha thalassemia major with significant hemoglobin Bart's, Causes nonimmune hydrops fetalis, usually fatal, Two genes defective (mild to moderate decrease in beta globin synthesis), Variable degrees of severity of symptoms of thalassemia major, Two genes defective (severe decrease in beta globin synthesis), Abdominal swelling, growth retardation, irritability, jaundice, pallor, skeletal abnormalities, splenomegaly; requires lifelong blood transfusions, MCV (abnormal if < 80 fl in adults; < 70 fl in children six months to six years of age; and < 76 fl in children seven to 12 years of age), Mentzer index for children (MCV/red blood cell count), Increased HbA2, reduced HbA, and probably increased HbF, Adults: normal Newborns: may have HbH or Hb Bart's. Normal Results In adults, these are normal percentages of different hemoglobin molecules: HbA: 95% to 98% (0.95 to 0.98) HbA2: 2% to 3% (0.02 to 0.03) HbE: Absent HbF: 0.8% to 2% (0.008 to 0.02) HbS: Absent HbC: Absent In infants and children, these are normal percentage of HbF molecules: HbF (newborn): 50% to 80% (0.5 to 0.8) HbF (6 months): 8% Hemoglobin. This is because the volume of blood has to increase up to 50% in order for a body to provide the baby with all necessary nutrients. [9, 10]. Hemoglobin is the protein inside red blood cells responsible for transporting oxygen to your tissues and organs. In the newborn period, if the electrophoresis shows Hb Bart's or HbH, the infant has alpha thalassemia. Imbalances of globin chains cause hemolysis and impair erythropoiesis. Hemoglobin Electrophoresis,HGB Variants,HB S,Hemoglobin F,HGB Evaluation,Hemoglobin S,HB A2,HGB Electrophoresis,Hemoglobin Variants,Hemoglobin A2,Thalassemia Screen,HB F. LOINC Codes, Performing Laboratory . 13.7. Hemoglobin type levels are a way of evaluating hemoglobin, a protein in your red blood cells. Hemoglobin F still makes up the majority of hemoglobin in newborns. A hemoglobin test measures the amount of hemoglobin in your red blood cells. Soon after birth it gets converted to Hemoglobin A. Hemoglobin S, C, E and M These are abnormal hemoglobin variants. Normal types of hemoglobin include: Hemoglobin (Hgb) A, the most common type of hemoglobin in healthy adults Hemoglobin (Hgb) F, fetal hemoglobin. They include: A hemoglobin electrophoresis test doesnt tell you about the amount of hemoglobin in your blood thats done in a complete blood count. What is the Hemoglobin electrophoresis test used for? Copyright 2023 American Academy of Family Physicians. National Human Genome Research Institute. Comment sent successfully! . For example, Dr. Abimola Farinde on Medscape says that normal hemoglobin levels for men could be as low as 13.0 g/dL. disopyramide phosphate . If you had a hemoglobin electrophoresis test because you had certain symptoms, you should call your healthcare provider to review your results. A thalassemia results in an abnormally low quantity of a globin chain. Most patients with abnormal hemoglobins have no symptoms but could potentially transmit the gene to their fetus. Your baby may feel a little pinch when the heel is poked, and a small bruise may form at the site. Reference or 'normal' range: determined by testing non-diseased individuals, usually middle 95% of values. Hemoglobin electrophoresis measures hemoglobin levels and looks for abnormal types of hemoglobin. If the synthesis from both genes is severely reduced or absent, the person has beta thalassemia major, also known as Cooley anemia. This study aimed to determine whether Hb levels in early pregnancy were associated with the risk of gestational diabetes mellitus (GDM), pre-eclampsia (PE . It also transports carbon dioxide out of your cells and. 11.9. Hemoglobin electrophoresis test uses electric current to segregate different hemoglobin proteins in blood. The cookie is set by GDPR cookie consent to record the user consent for the cookies in the category "Functional". Newborn Screening Tests for Your Baby; [cited 2020 Jan 10]; [about 3 screens]. Your lab report should show the range that your lab uses for each test. Transfusion-dependent patients will develop iron overload and require chelation therapy to remove the excess iron. Your blood gets its red color from a protein called hemoglobin which also helps transfer oxygen to different cells of your body. Bone marrow aspiration and biopsy are generally not. You can learn more about how we ensure our content is accurate and current by reading our. Pressure is applied on the site and then a bandage is put. A single gene deletion results in alpha thalassemia silent carrier status, which is asymptomatic with normal hematologic findings. Typically, healthcare providers obtain blood samples by drawing blood from a vein in your arm or hand. To analyze the types of hemoglobin present in a blood sample, the standard of care involves using two methods of analysis on each sample. Mosbys Diagnostic & Laboratory Test Reference. The lab technician may also perform a test known as hemoglobin electrophoresis. Microcytic hypochromic red cells with normal or borderline low Hb Diagnosed on HPLC or Hb electrophoresis - HbA2 > 3.5%, often elevated Hb F Red cell distribution width (RDW) is often normal Pre-pregnancy carrier testing of partner is important (Ensure parents have been tested if likely to have more children) If you have questions about your results, talk to your health care provider. The chamber is a machine that passes electrical currents through the sample. Hemoglobin F or Fetal hemoglobin- It is found in newborn kids and foetuses. The one gene defect, beta thalassemia trait (minor), is asymptomatic and results in microcytosis and mild anemia. The two-gene deletion causes alpha thalassemia trait (minor) with microcytosis and usually no anemia. Hemoglobin is a protein in red blood cells that carries oxygen from the lungs to the rest of the body. https://www.sciencedirect.com/topics/medicine-and-dentistry/hemoglobin-electrophoresis#:~:text=Haemoglobin%20electrophoresis%20provides%20qualitative%20analysis,quantitation%20of%20the%20Hb%20fractions. To screen for genetic conditions: People who have a family history of inherited anemias such as thalassemia or sickle cell anemia may choose to screen for these genetic disorders before having children. In other forms of sickle-cell disease, Hb levels tend to be higher. In addition to his current work, Dr. Amos is using his vast experience to launch Obie, a science-based app that offers personalized fertility advice. Abnormal cells on peripheral blood smear: The peripheral blood smear is perhaps the most inexpensive laboratory test that can accurately lead to a diagnosis of thalassemia. Beta-thalassemia causes symptoms related to anemia, including weak or brittle bones, and leads to problems with how your bones develop. This should go away quickly. If your test results show you or your child may have a serious illness, its easy to focus on the snapshot. The four-gene deletion results in significant production of hemoglobin Bart's (Hb Bart's), which has four gamma chains (gamma4). The vial is submitted for analysis. The major hemoglobin types have different electrical charges. The Korean journal of laboratory medicine. Learn more about laboratory tests, reference ranges, and understanding results. [6], Mass spectrometry, a newer analytical technology, may be used to identify highly unstable hemoglobins that may manifest clinically as hemolytic anemia or thalassemia. Our hospitals are equipped with advanced technology to perform a wide range of procedures and treatments. Homozygous sickle cell disease (Hb SS) and other sickle cell disease variants including sickle cell beta-thalassemia and hemoglobin SC disease are discussed separately. Hemoglobin abnormalities (2015, December 22), What is sickle cell disease? Oct 2011;31(4):238-243.: Caruso D, Crestani M, Mitro N, et al. FBS and PPBS values in pregnancy.. 88 and 144.. worried: Hi all most have undergone a GCT or GTT in pregnancy. Jacksonville (FL): The Nemours Foundation; c19952020. Hoffman: Hematology: Basic Principles and Practice. For persons with beta thalassemia intermedia, the decision to transfuse is a more subjective clinical assessment. Additionally, mass spectrometry may provide information on posttranslational modifications, such as oxidation and glycation. Hemoglobin C, S-C, and E Diseases; [updated 2019 Feb; cited 2020 Jan 10]; [about 2 screens]. Sickle cell can be diagnosed by a blood test. A normal serum lead level excludes lead poisoning. Aga Khan lab test directory with complete lab test details regarding biochemistry, blood bank, haematology, histopathology, & molecular pathology. Hemoglobin electrophoresis has been superseded by more rapid, sensitive, and quantitative methods of hemoglobin separation.