Folate (Fol) Atellica IM Analyzer

Folate (Fol)Current Revision and Datea Rev. 04, 2020-11

Product Name Atellica IM Folate (Fol) 10995572(140 tests)

10995573(700 tests)

Abbreviated Product Name Atellica IM FolTest Name/ID FolSystems Atellica IM AnalyzerMaterials Required but Not Provided Atellica IM Fol DTT/REL 10995576

Atellica IM APW1 10995458

Optional Materials Atellica IM RBC Fol 10995440

Atellica IM Fol DIL 10995574

Atellica IM Fol MCM 10995575

Specimen Types Serum, dipotassium EDTA whole blood, heparinized whole bloodSample Volume 100 µLMeasuring Interval Serum: 0.56–24.00 ng/mL (1.27–54.36 nmol/L)

RBC Hemolysate: 0.98–17.51 ng/mL (2.22–39.66 nmol/L)a A vertical bar in the page margin indicates technical content that differs from the previous version.

Intended UseThe Atellica® IM Folate (Fol) assay is for in vitro diagnostic use in the quantitativedetermination of folate in human serum and red blood cells using the Atellica® IM Analyzer.Folic acid measurements are used in the diagnosis and treatment of anemias.

Summary and ExplanationFolates are compounds of pteroylglutamic acid (PGA) that function as coenzymes in metabolicreactions involving the transfer of single-carbon units from a donor to a recipient compound.Folate, with vitamin B12, is essential for DNA synthesis, which is required for normal red bloodcell maturation.1 Humans obtain folate from dietary sources including fruits, green and leafyvegetables, yeast, and organ meats.2 Folate is absorbed through the small intestine and storedin the liver.

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Low folate intake, malabsorption as a result of gastrointestinal diseases, pregnancy, and drugssuch as phenytoin are causes of folate deficiency.3 Folate deficiency is also associated withchronic alcoholism.4 Folate and vitamin B12 deficiency impair DNA synthesis, causingmacrocytic anemias. These anemias are characterized by abnormal maturation of red bloodcell precursors in the bone marrow, the presence of megaloblasts, and decreased red bloodcell survival.1

Since both folate and vitamin B12 deficiency can cause macrocytic anemia, appropriatetreatment depends on the differential diagnosis of the deficiency. Serum folate measurementprovides an early index of folate status.2 However, folate is much more concentrated in redblood cells than in serum so the red blood cell folate measurement more closely reflects tissuestores.4 Red blood cell folate concentration is considered the most reliable indicator of folatestatus.2

Principles of the ProcedureThe Atellica IM Fol assay is a competitive immunoassay using direct chemiluminescenttechnology. Folate in the patient sample competes with acridinium-ester-labeled folate in theLite Reagent for a limited amount of biotin-labeled folate binding protein. Biotin-labeled folatebinding protein binds to avidin that is covalently coupled to paramagnetic particles in the SolidPhase. In the Atellica IM Fol assay, the sample is pretreated to release the folate fromendogenous binding proteins in the sample.An inverse relationship exists between the amount of folate present in the patient sample andthe amount of relative light units (RLUs) detected by the system.

ReagentsMaterial Description Storage Stabilitya

Atellica IM Fol ReadyPack® primary reagent packLite Reagent9.1 mL/reagent packFolate labeled with acridinium ester (~9.8 ng/mL) in buffer;bovine serum albumin; sodium azide (0.1%); preservativesSolid Phase18.2 mL/reagent packPurified avidin (~20 µg/mL) covalently coupled toparamagnetic particles in buffer; human serum albumin;preservativesFolate Binding Protein9.1 mL/reagent packPurified folate binding protein (~1.0 µg/mL) covalentlycoupled to biotin in buffer; bovine serum albumin;preservatives

Unopened at 2–8°C Until expiration dateon product

Onboard 14 days

Atellica IM Fol CAL3.0 mL/vial; lyophilizedAfter reconstitution, low or high levels of N-5-methyltetrahydrofolic acid; buffer; human serum albumin;sodium azide (< 0.1%); preservatives

Lyophilized at 2–8°C Until expiration dateon product

Reconstituted at 2–8°C 7 daysReconstituted at roomtemperature

8 hours

Reconstituted at≤ -20°C

28 days

Atellica® SampleHandlerb

Fol Atellica IM Analyzer

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Material Description Storage Stabilitya

Atellica IM Fol DTT/Releasing Agentc

DTT8.0 mL/vialDithiothreitol (~95 mg/mL in liquid form)Releasing Agent4.0 mL/vialSodium hydroxide (~1.1 N)

At 2–8°C Until expiration dateon product

Onboard in an ancillaryreagent packd

108 hours

Atellica IM RBC Fole, f

RBC Folate Ascorbic AcidLyophilized ascorbic acid (~0.30 g/vial)RBC Folate Ascorbic Acid Diluent30.0 mL/vialBovine serum albumin; buffer; sodium azide (< 0.1%);preservatives


Reconstituted at 2–8°C 30 days

Atellica IM Fol DIL ReadyPack ancillary reagent packe

10.0 mL/reagent packHuman plasma; sodium azide (< 0.1%); preservatives


Onboard 28 days

Atellica IM APW1 ReadyPack ancillary reagent packc

25.0 mL/pack0.4 N sodium hydroxide


Onboard 14 daysa Refer to Storage and Stability.b Refer to the supplementary document "Atellica Sample Handler Calibrator and QC Storage and Stability" for

information about storage and stability of materials in the Cal‑QC tube storage area.c Refer to Materials Required but Not Provided.d Refer to Preparing the Reagents.e Refer to Optional Materials.f Refer to Preparing the Red Blood Cell Hemolysate in the Preparing the Samples section.

Warnings and PrecautionsFor in vitro diagnostic use.For Professional Use.CAUTIONFederal (USA) law restricts this device to sale by or on the order of a licensed healthcareprofessional.Safety data sheets (SDS) available on siemens.com/healthineers.

H290, H314P280, P310,P305+P351+P338,P301+P330+P331,P303+P361+P353,P390, P501

Danger!May be corrosive to metals. Causes severe skin burns and eye damage.Wear protective gloves/protective clothing/eye protection/face protection.Immediately call a POISON CENTER or doctor/physician. IF IN EYES: Rinsecautiously with water for several minutes. Remove contact lenses, ifpresent and easy to do. Continue rinsing. IF SWALLOWED: rinse mouth.Do NOT induce vomiting. IF ON SKIN (or hair): Remove/Take offimmediately all contaminated clothing. Rinse skin with water/shower.Absorb spillage to prevent material damage. Dispose of contents andcontainer in accordance with all local, regional, and national regulations.Contains: sodium hydroxide (in Atellica IM Fol DTT/Releasing Agent)

Atellica IM Analyzer Fol

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http://siemens.com/healthineers

H290, H319, H315P280, P264,P305+P351+P338,P310, P390, P501

Warning!May be corrosive to metals. Causes serious eye irritation. Causes skinirritation.Wear protective gloves/protective clothing/eye protection/face protection.Wash hands thoroughly after handling. IF IN EYES: Rinse cautiously withwater for several minutes. Remove contact lenses, if present and easy todo. Continue rinsing. Immediately call a POISON CENTER or doctor/physician. Absorb spillage to prevent material damage. Dispose ofcontents and container in accordance with all local, regional, andnational regulations.Contains: sodium hydroxide (in Atellica IM APW1)

H412P273, P501

Harmful to aquatic life with long lasting effects.Avoid release to the environment. Dispose of contents and container inaccordance with all local, regional, and national regulations.Contains: sodium azide (in Atellica IM Fol CAL)

CAUTION POTENTIAL BIOHAZARDContains human source material. Each donation of human blood or blood component wastested by FDA-approved methods for the presence of antibodies to human immunodeficiencyvirus type 1 (HIV‑1) and type 2 (HIV‑2), as well as for hepatitis B surface antigen (HBsAg) andantibody to hepatitis C virus (HCV). The test results were negative (not repeatedly reactive). Notest offers complete assurance that these or other infectious agents are absent; this materialshould be handled using good laboratory practices and universal precautions.5–7

CAUTIONThis device contains material of animal origin and should be handled as a potential carrier andtransmitter of disease.Contains sodium azide as a preservative. Sodium azide can react with copper or lead plumbingto form explosive metal azides. On disposal, flush reagents with a large volume of water toprevent buildup of azides. Disposal into drain systems must be in compliance with prevailingregulatory requirements.Dispose of hazardous or biologically contaminated materials according to the practices of yourinstitution. Discard all materials in a safe and acceptable manner and in compliance withprevailing regulatory requirements.Note For information about reagent preparation, refer to Preparing the Reagents in theProcedure section.Note For information about calibrator preparation, refer to Preparing the Calibrators.

Storage and StabilityStore reagents in an upright position. Protect the product from heat and light sources.Unopened reagents are stable until the expiration date on the product when stored at 2–8°C.Store calibrators in an upright position. Lyophilized calibrators are stable until the expirationdate on the product when stored at 2–8°C. Reconstituted calibrators are stable for 7 days at2–8°C or for 8 hours at room temperature. Freeze reconstituted product at ≤ -20°C for up to28 days.Store Atellica IM Fol DTT/Releasing Agent in an upright position. Unopened Atellica IM Fol DTT/Releasing Agent is stable until the expiration date on the product when stored at 2–8°C.


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Store Atellica IM RBC Fol in an upright position. Unopened Atellica IM RBC Fol is stable until theexpiration date on the product when stored at 2–8°C. Reconstituted material is stable for30 days at 2–8°C.Store Atellica IM Fol DIL in an upright position. Unopened Atellica IM Fol DIL is stable until theexpiration date on the product when stored at 2–8°C.Store Atellica IM APW1 in an upright position. Unopened Atellica IM APW1 is stable until theexpiration date on the product when stored at 2–8°C.Do not use products beyond the expiration date printed on the product labeling.

Onboard StabilityAtellica IM Fol ReadyPack primary reagent packs are stable onboard the system for 14 days.Discard reagents at the end of the onboard stability interval.Note Refer to the supplementary document "Atellica Sample Handler Calibrator and QCStorage and Stability" for information about storage and stability of materials in the Cal‑QCtube storage area.Atellica IM Fol DTT/Releasing Agent is stable onboard the system for 108 hours.Atellica IM Fol DIL is stable onboard the system for 28 days.Atellica IM APW1 is stable onboard the system for 14 days.Do not use products beyond the expiration date printed on the product labeling.

Specimen Collection and HandlingSerum and red blood cell hemolysate (prepared from dipotassium EDTA or heparinized wholeblood) are the recommended sample types for this assay.Note Folates are light-sensitive. Minimize exposure to light during sample handling andstorage.8

Collecting the Specimen• Observe universal precautions when collecting specimens. Handle all specimens as if they

are capable of transmitting disease.7

• Follow recommended procedures for collection of diagnostic blood specimens byvenipuncture.9

• Follow the instructions provided with your specimen collection device for use andprocessing.10

• Allow blood specimens to clot completely before centrifugation.6

• Keep tubes capped at all times.6

Storing the Specimen• Store serum samples at room temperature for no longer than 8 hours.• Tightly cap and refrigerate specimens at 2–8°C if the assay is not completed within

8 hours.• Freeze serum samples at ≤ -20°C if the assay is not completed within 48 hours.


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• Freeze serum samples only 1 time and mix thoroughly after thawing. Frozen specimenscan remain frozen for up to 30 days. Do not store in a frost-free freezer. If serum sampleswill be stored for longer than 30 days, then they must be frozen at ≤ -80°C.

• If testing is not done within 24 hours for whole blood specimens, determine thehematocrit and freeze the whole blood specimen or hemolysate. Frozen whole bloodspecimens can be stored at -20°C for up to 2 months. Sample hemolysates prepared withthe reconstituted RBC Folate Ascorbic Acid can be stored at -20°C for up to 3 months. Donot store whole blood specimens or hemolysates in a frost-free freezer. Freeze specimensonly 1 time and mix thoroughly after thawing.

The handling and storage information provided here is based on data or referencesmaintained by the manufacturer. It is the responsibility of the individual laboratory to use allavailable references and/or its own studies when establishing alternate stability criteria tomeet specific needs.

Transporting the SpecimenPackage and label specimens for shipment in compliance with applicable federal andinternational regulations covering the transport of clinical specimens and etiological agents.

Preparing the SamplesThis assay requires 100 µL of sample for a single determination. This volume does not includethe unusable volume in the sample container or the additional volume required whenperforming duplicates or other tests on the same sample. For information about determiningthe minimum required volume, refer to the online help.The sample volume required to perform onboard dilution differs from the sample volumerequired to perform a single determination. Refer to Dilutions.Note Do not use specimens with apparent contamination.Before placing samples on the system, ensure that samples are free of:• Bubbles or foam.• Fibrin or other particulate matter.Note Remove particulates by centrifugation according to CLSI guidance and the collectiondevice manufacturer’s recommendations.6

Note For a complete list of appropriate sample containers, refer to the online help.Preparing the Red Blood Cell Hemolysate

Note Folates are light sensitive. Minimize exposure to light during sample handling andstorage.8

1. Reconstitute the RBC Folate Ascorbic Acid by adding the contents of 1 vial of RBC FolateAscorbic Acid Diluent to the lyophilized RBC Folate Ascorbic Acid.

2. Let the reconstituted mixture stand at room temperature for 15 minutes and mix byinverting the bottle occasionally.

3. Collect the sample in a tube containing heparin or dipotassium EDTA.4. Invert the sample several times to mix.5. Determine and record the hematocrit.6. Dispense 1.0 mL of reconstituted RBC Folate Ascorbic Acid into a test tube or sample cup.7. Add 50 µL of the sample into the RBC Folate Ascorbic Acid.8. Cap and invert the tube several times or vortex gently to mix. Avoid foaming.


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9. Let the hemolysate stand protected from light, at room temperature, for a minimum of90 minutes, but less than 3 hours.

10. Do not mix the hemolysate again before placing the sample on the system.Note Do not dilute the RBC hemolysate. Freeze the hemolysate at ≤ -20°C immediately iftesting cannot be completed within 4 hours from the time you finish preparing thehemolysate. Hemolysates can be stored at ≤ -20°C for up to 3 months. Do not store in a frost-free freezer.If the hemolysate is frozen, thaw the hemolysate and mix it by inverting the tube severaltimes. Let the hemolysate stand for 30 minutes at room temperature before testing. Test thehemolysate within 3 hours after thawing.

ProcedureMaterials Provided

The following materials are provided:

ContentsNumber ofTests

10995572 1 ReadyPack primary reagent pack containing Atellica IM Fol Lite Reagent,Solid Phase, and Folate Binding ProteinAtellica IM Fol master curve and test definition 2 vials Atellica IM Fol CAL low calibrator 2 vials Atellica IM Fol CAL high calibrator Atellica IM Fol CAL calibrator lot-specific value sheet

140

10995573 5 ReadyPack primary reagent packs containing Atellica IM Fol Lite Reagent,Solid Phase, and Folate Binding ProteinAtellica IM Fol master curve and test definition 2 vials Atellica IM Fol CAL low calibrator 2 vials Atellica IM Fol CAL high calibrator Atellica IM Fol CAL calibrator lot-specific value sheet

700

Materials Required but Not ProvidedThe following materials are required to perform this assay, but are not provided:

Description

Atellica IM Analyzera

10995576 Atellica IM Fol DTT/REL (releasingagent)

3 x 8.0 mL/vial DTT3 x 4.0 mL/vial Releasing Agent 3 empty ReadyPack ancillary reagent packs

10995458 Atellica IM APW1 (probe wash) 2 ReadyPack ancillary reagent packs containing 25.0 mL/pack

a Additional system fluids are required to operate the system: Atellica IM Wash, Atellica IM Acid, Atellica IM Base,and Atellica IM Cleaner. For system fluid instructions for use, refer to the Document Library.


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Optional MaterialsThe following materials may be used to perform this assay, but are not provided:

Description10995440 Atellica IM RBC Fol (ascorbic acid/diluent) 4 x 0.30 g/vial Ascorbic Acid

4 x 30.0 mL/vial Ascorbic Acid Diluent

10995574 Atellica IM Fol DIL (diluent) 2 ReadyPack ancillary reagent packs containing10.0 mL/pack

10995575 Atellica IM Fol MCM (master curve material) 6 x 1.0 mL levels of master curve material

Assay ProcedureThe system automatically performs the following steps:1. Dispenses 100 µL of sample into a cuvette.2. Dispenses 35 µL of DTT/Releasing Agent, then incubates for 5 minutes at 37°C.3. Dispenses 65 µL of Folate Binding Protein and 130 µL of Solid Phase, then incubates for

5 minutes at 37°C.4. Dispenses 65 µL of Lite Reagent, then incubates for 3 minutes at 37°C.5. Separates, aspirates, then washes the cuvettes with Atellica IM Wash.6. Dispenses 300 μL each of Atellica IM Acid and Atellica IM Base to initiate the

chemiluminescent reaction.7. Reports results.

Preparing the ReagentsAll reagents are liquid and ready to use, with the exception of the Atellica IM Fol DTT/ReleasingAgent. Before loading primary reagent packs onto the system, mix them by hand and visuallyinspect the bottom of the reagent pack to ensure that all particles are resuspended. Forinformation about preparing the reagents for use, refer to the online help.

Preparing the DTT/Releasing AgentNote Careful preparation of DTT/Releasing Agent is required to obtain accurate and consistentresults since the absolute amount of DTT delivered to each test can affect results. The preparedDTT/Releasing Agent must be loaded onto the system immediately and used within 108 hoursafter preparation.1. Carefully transfer the contents of 1 vial of Releasing Agent into 1 vial of DTT. For

convenience, the Releasing Agent can be poured or transferred by pipette into the DTTvial.

2. Firmly screw the cap on the DTT vial and invert the vial several times to mix.3. Pour or pipette the entire contents of the DTT vial into the disposable ancillary reagent

pack provided.4. Place a pack seal on the disposable ancillary reagent pack. Ensure that the seal is centered

over the openings of the pack, and press firmly on the adhesive portion of the seal.Note DTT/Releasing Agent ReadyPack ancillary reagent packs are lot-number-specific. Do notuse packs from one lot of DTT/Releasing Agent with any other lot of DTT/Releasing Agent.


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Preparing the SystemEnsure that the system has sufficient reagent packs loaded in the reagent compartment. Thesystem automatically mixes reagent packs to maintain homogeneous suspension of thereagents. For information about loading reagent packs, refer to the online help.For automated dilutions, ensure that Atellica IM Fol DIL is loaded in the reagent compartment.

Master Curve DefinitionBefore initiating calibration on each new lot of reagent, load the assay master curve and testdefinition values by scanning the 2D barcodes. For loading instructions, refer to theonline help.

Performing CalibrationFor calibration of the Atellica IM Fol assay, use the calibrators provided with each kit.Do not pour the calibrators back into the vials after calibration because evaporation couldoccur, which may affect performance.Do not refill calibrator sample cups when the contents are depleted. If required, dispense freshcalibrators.

Calibration FrequencyPerform a calibration if one or more of the following conditions exist:• When changing lot numbers of primary reagent packs.• At the end of the lot calibration interval, for a specified lot of calibrated reagent on the

system.• At the end of the pack calibration interval, for calibrated reagent packs on the system.• When indicated by quality control results.• After major maintenance or service, if indicated by quality control results.At the end of the onboard stability interval, replace the reagent pack on the system with a newreagent pack. Recalibration is not required, unless the lot calibration interval is exceeded.

Stability Interval DaysLot Calibration 14Pack Calibration 7Reagent Onboard Stability 14

For information about lot calibration and pack calibration intervals, refer to the online help.Follow government regulations or accreditation requirements for calibration frequency.Individual laboratory quality control programs and procedures may require more frequentcalibration.

Preparing the CalibratorsPrepare calibrators using the following steps:1. Add 3.0 mL of special reagent water into each vial using a precision pipet. Replace cap.

Note For information about special reagent water requirements, refer to the online help.2. Let the vials stand for 15–20 minutes at room temperature to allow the lyophilized

material to dissolve.3. Gently mix and invert the vials to ensure homogeneity of the material.


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4. For extended storage, aliquot and seal tightly. Store reconstituted material according tostability limits specified in Storing the Specimen. Do not store in a frost-free freezer.Note Before using frozen calibrators, allow the material to completely thaw. Gently mixand invert the vials to ensure homogeneity of the material. Use immediately and discardany remaining material.

Note Use calibrators within the stability limits specified in Storing the Specimen and discardany remaining material.

Calibration ProcedureThe required sample volume for testing depends on several factors. For information aboutsample volume requirements, refer to the online help.Use the following lot‑specific materials to perform calibration:• For the master curve and assay test definitions, refer to the lot‑specific master curve and

test definition sheet provided with the assay reagents.• Calibrators provided in an assay kit must only be used with reagents from that assay kit lot.

Do not use calibrators from one assay kit with reagents from a different assay kit lot.• For the calibrator definitions, refer to the lot‑specific value sheet provided with

the calibrator materials.• Generate lot‑specific barcode labels to use with the calibrator samples.For instructions about how to perform the calibration procedure, refer to the online help.

Performing Quality ControlFor quality control of the Atellica IM Fol assay, use an appropriate quality control material ofknown analyte concentration with at least 2 levels (low and high) at least once during eachday that samples are analyzed. Use the quality control material in accordance with the qualitycontrol instructions for use.A satisfactory level of performance is achieved when the analyte values obtained are withinthe expected control interval for the system or within your interval, as determined by anappropriate internal laboratory quality control scheme. Follow your laboratory’s quality controlprocedures if the results obtained do not fall within the acceptable limits. For informationabout entering quality control definitions, refer to the online help.Follow government regulations or accreditation requirements for quality control frequency.Individual laboratory quality control programs and procedures may require more frequentquality control testing.Test quality control samples after a successful calibration.

Taking Corrective ActionIf the quality control results do not fall within the assigned values, do not report results.Perform corrective actions in accordance with established laboratory protocol. For suggestedprotocol, refer to the online help.

ResultsCalculation of Results

The system determines the result using the calculation scheme described in the online help.The system reports results in ng/mL (common units) or nmol/L (SI units), depending on theunits defined when setting up the assay.Conversion formula: 1 ng/mL = 2.265 nmol/L


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For information about results outside the specified measuring interval, refer to MeasuringInterval.

Atellica IM Test OptionsThe Atellica IM Fol assay can be used for the following test options:• Fol = system test generated serum folate concentration• rFOL = predefined ratio test on the system for system-generated hemolysate folate value• rbcFOL = predefined ratio test on the system for system-generated hemolysate RBC folate

value (requires off-system hematocrit value)Note For information about creating the off-system hematocrit test, refer to the onlinehelp.

• Manually calculated RBC Folate value using the rFOL predefined ratio value generated bythe system for the hemolysate and the off-system hematocrit value

• Corrected Red Blood Cell Folate = system or manually calculated corrected RBC FolateFor information about entering the off-system and ratio test definitions, refer to the onlinehelp.Note Use serum samples with the Fol test; use RBC hemolysate samples with rFOL or rbcFOLpredefined ratio tests

Manual Calculation for Red Blood Cell FolateUse this procedure to manually calculate RBC folate values using an off-system hematocritvalue and the result from the rFOL test.1. Prepare the RBC hemolysate as described in Preparing the Red Blood Cell Hemolysate.2. Order the rFOL predefined ratio test.

Note The rFOL predefined ratio test must use RBC hemolysate sample. Do not use othersample types.

3. Multiply the folate result for the hemolysate by 21 (a 1:21 dilution was made whenpreparing the RBC hemolysate). This value represents the folate concentration of wholeblood in ng/mL.

4. Divide this result by the hematocrit, and multiply by 100 to adjust for the hematocrit,which is a percentage.

RBC folate (ng/mL) = Folate result for hemolysate, (ng/mL) x 21

x 100hematocrit

Example:Hemolysate folate value = 5.7 ng/mLHematocrit = 43

RBC folate (ng/mL) = 5.7 x 21

x 100 = 27843

Red Blood Cell Folate using the predefined system ratio test (rbcFOL)1. Prepare the RBC hemolysate as described in Preparing the Red Blood Cell Hemolysate.2. Order rbcFOL from the list of predefined ratio tests.

Note The rbcFOL predefined ratio test must use RBC hemolysate sample. Do not use othersample types.


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3. Manually enter the hematocrit value for the sample.4. The system will display the calculated red blood cell folate value.

Corrected Red Blood Cell FolateIn most cases the serum folate values are very small compared to red blood cell folate values.However, occasionally serum folate values can be elevated. If the serum folate value is highand the red blood cell folate concentration is low, calculate the corrected RBC folate valueaccording to the following equation:

For information about entering the off-system and ratio test definitions, refer to the onlinehelp.

DilutionsThe measuring interval for serum is 0.56–24.00 ng/mL (1.27–54.36 nmol/L). For informationabout dilution options, refer to the online help.Serum samples with folate levels > 24.00 ng/mL (54.36 nmol/L) must be diluted and retestedto obtain accurate results.Note Do not dilute the RBC hemolysate.For automated dilutions, ensure that Atellica IM Fol DIL is loaded in the reagent compartment.Ensure that sufficient sample volume is available to perform the dilution and that theappropriate dilution factor is selected when scheduling the test, as indicated in the tablebelow.For automatic dilutions, enter a dilution setpoint ≤ 24 ng/mL (54.36 nmol/L).

Sample Dilution Sample Volume (µL)Serum 1:2 100

Interpretation of ResultsResults of this assay should always be interpreted in conjunction with the patient’s medicalhistory, clinical presentation, and other findings.

LimitationsThe following information pertains to limitations of the assay:• Hemolysis significantly increases folate values in serum due to the high folate

concentrations found in red blood cells.• Methotrexate and leucovorin interfere with folate measurement because these drugs

cross-react with folate binding proteins.


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Expected ValuesThe reagent formulations used on the Atellica IM Analyzer are the same as those used on theADVIA Centaur® system. Expected values were established using the ADVIA Centaur systemand confirmed by assay comparison. Refer to Assay Comparison.To determine the reference intervals for serum and RBC folate, data were obtained using 370and 286 samples, respectively. For sample results in the indeterminate range[3.38–5.38 ng/mL (7.64–12.19 nmol/L)], clinical results and other diagnostic protocols shouldsupplement folate results.

Category NaMedian(ng/mL)

Range(ng/mL)

Median(nmol/L)

Range(nmol/L)

Serum folateDeficientc 65 1.54 < 0.56–3.37 3.49 < 1.27–7.63

Indeterminated 3.38–5.38 7.64–12.19

Normal 305 12.51 > 5.38b 28.34 > 12.19

RBC folateNormal 286 425 280–791 963 634–1792

a Number of samples.b Inner 97.5% of the distribution of apparently healthy individuals.c Diagnosed by bone and/or peripheral blood smear pathology and other criteria including:• megaloblastic anemia• folate-deficient diet• malabsorption• alcoholism• Tropical Sprue• abnormal blood parameters including mean corpuscular volume (MCV), mean corpuscular hemoglobin

(MCH), and hematocrit (HCT)d Range between deficient and normal range.

Laboratories should consider these expected values as guidelines only. The data were obtainedon apparently healthy males and females from the United States. Because of populationdemographic factors, diet, and assay methods, each laboratory should determine its ownexpected values for the diagnostic evaluation of patient results.11

Performance CharacteristicsThe reagent formulations used on the Atellica IM Analyzer are the same as those used on theADVIA Centaur® system. Some performance characteristics for the Atellica IM assay wereestablished using the ADVIA Centaur system.

Measuring IntervalThe Atellica IM Fol assay provides results from 0.56–24.00 ng/mL (1.27–54.36 nmol/L) forserum. Report results below the measuring interval as < 0.56 ng/mL (1.27 nmol/L). Whensample results exceed the measuring interval, refer to Dilutions.The Atellica IM Fol assay provides results from 0.98–17.51 ng/mL (2.22–39.66 nmol/L) for RBChemolysates. Do not dilute the RBC hemolysates.


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SpecificityCross-reactivity was determined in accordance with CLSI Document EP07‑A215 using theAtellica IM Analyzer. Samples were spiked with the compounds listed below at the indicatedlevels. The following results were obtained:

SubstanceSubstance Test Concentration(ng/mL)

Analyte Concentrationng/mL (nmol/L) Cross-Reactivity (%)

Amethopterin 150 0.00 (0.00) 0.9150 3.31 (7.50) 1.4150 9.46 (21.43) 1.4

Aminopterin 75 0.00 (0.00) 0.075 3.30 (7.47) 0.175 9.31 (21.09) -0.2

Folinic Acid 25 0.00 (0.00) 0.125 3.41 (7.72) -1.825 9.29 (21.04) -3.9

Assay results obtained at individual laboratories may vary from the data presented.

Detection CapabilityDetection capability was determined in accordance with CLSI Document EP17‑A2.12 The assayis designed to have a limit of blank (LoB) ≤ 0.40 ng/mL (0.91 nmol/L), a limit of detection(LoD) ≤ 0.70 ng/mL (1.59 nmol/L), and a limit of quantitation (LoQ) ≤ 0.70 ng/mL(1.59 nmol/L).Representative detection capability data are shown below. Assay results obtained at individuallaboratories may vary from the data presented.The LoB corresponds to the highest measurement result that is likely to be observed for ablank sample. The LoB of the Atellica IM Fol assay is 0.19 ng/mL (0.43 nmol/L) for serum and0.00 ng/mL (0.00 nmol/L) for RBC hemolysate.The LoD corresponds to the lowest concentration of folate that can be detected with aprobability of 95%. The LoD for the Atellica IM Fol assay is 0.38 ng/mL (0.86 nmol/L) for serum,and was determined using 540 determinations, with 480 blank and 60 low-level replicates,and an LoB of 0.19 ng/mL (0.43 nmol/L). The LoD for Atellica IM Fol assay is 0.21 ng/mL(0.48 nmol/L) for RBC hemolysate and was determined using 473 determinations, with120 blank and 353 low-level replicates, and an LoB of 0.00 ng/mL (0.00 nmol/L).The LoQ corresponds to the lowest amount of folate in a sample at which the within-laboratory CV is ≤ 20%. The LoQ of the Atellica IM Fol assay is 0.56 ng/mL (1.27 nmol/L), andwas determined using multiple patient samples in the interval 0.27–1.25 ng/mL(0.61–2.83 nmol/L). All samples were assayed in 10 replicates, 2 runs per day using 3 reagentlots, over a period of 5 days.

PrecisionPrecision was determined in accordance with CLSI Document EP05‑A3.13 Samples wereassayed on an Atellica IM Analyzer in duplicate in 2 runs per day for 20 days. The assay wasdesigned to have within-laboratory precision for serum ≤ 0.21 ng/mL (0.48 nmol/L) SD forsamples < 1.80 ng/mL (4.08 nmol/L) and ≤ 11% CV for samples from 1.90–24.00 ng/mL(4.30–54.36 nmol/L).


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The assay was designed to have within-laboratory precision for red blood cell hemolysate≤ 14.50 ng/mL (32.84 nmol/L) SD for samples < 130.99 ng/mL (296.69 nmol/L) and ≤ 11% CVfor samples ≥ 131.00 ng/mL (296.72 nmol/L).

Sample Type Na

Mean

Repeatability

Within-Laboratory Precision

(ng/mL) (nmol/L)SDb

CVc

(%)SD

CV(%)(ng/mL) (nmol/L) (ng/mL) (nmol/L)

Serum A 80 1.42 3.22 0.05 0.11 N/Ad 0.08 0.18 N/A

Serum B 80 4.13 9.35 0.10 0.23 2.4 0.24 0.54 5.9Serum C 80 6.19 14.02 0.18 0.41 2.9 0.36 0.82 5.9Serum D 80 9.23 20.91 0.24 0.54 2.6 0.6 1.36 6.5Serum Control 1 80 2.82 6.39 0.09 0.20 3.3 0.17 0.39 6.1Serum Control 2 80 5.43 12.30 0.14 0.32 2.5 0.35 0.79 6.4

Whole BloodSample A

80 94.89 214.93 4.56 10.33 N/A 6.55 14.84 N/A

Whole BloodSample B

80 153.11 346.79 4.40 9.97 2.9 11.21 25.39 7.3

Whole BloodSample C

80 362.58 821.24 9.82 22.24 2.7 19.99 45.28 5.5

Whole BloodSample D

80 563.94 1277.32 19.34 43.81 3.4 35.18 79.68 6.2

Whole BloodSample E

80 899.24 2036.78 45.66 103.42 5.1 62.92 142.51 7.0

Whole BloodControl 1

80 77.24 174.95 2.68 6.07 N/A 5.48 12.41 N/A

Whole BloodControl 2

80 257.52 583.28 6.51 14.75 2.5 16.84 38.14 6.5

a Number of samples tested.b Standard deviation.c Coefficient of variation.d Not applicable; there is no CV requirement at these concentrations.

Collection Tube ComparisonRed Blood Cell Folate

Specimen equivalency was determined using weighted Deming linear regression inaccordance with CLSI Document EP09‑A3.14 The following results were obtained:

Specimen (y)Reference Specimen(x) Regression Equation Sample Interval Na

Dipotassium EDTAwhole blood

Lithium heparin wholeblood

y = 1.02x - 32.86 ng/mL(y = 1.02x - 74.43 nmol/L)

432.70–1103.69 ng/mL(980.07–2499.86 nmol/L)

90

a Number of samples tested.


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Assay ComparisonThe Atellica IM Fol assay is designed to have a correlation coefficient of ≥ 0.90 and a slope of1.0 ± 0.1 for serum when compared to the ADVIA Centaur Folate assay. Assay comparison wasdetermined using the weighted Deming regression model in accordance with CLSI DocumentEP09‑A3.14 The following results were obtained:

Specimen Comparative Assay (x) Regression Equation Sample Interval Na rb

Serum ADVIA Centaur Folate y = 0.94x - 0.01 ng/mL(y = 0.94x - 0.02 nmol/L)

0.64–22.78 ng/mL(1.45–51.60 nmol/L)

105 0.99

RBC hemolysate ADVIA Centaur Folate y = 1.06x - 2.52 ng/mL(y = 1.06x - 5.71 nmol/L)

229.66–2264.56 ng/mL(520.18–5129.23 nmol/L)

100 0.93

a Number of samples tested.b Correlation coefficient.

Agreement of the assays may vary depending on the study design, comparative assay, andsample population used. Assay results obtained at individual laboratories may vary from thedata presented.

InterferencesInterference testing was performed in accordance with CLSI Document EP07‑A215 using theAtellica IM Analyzer.The following substances were added to serum samples containing different concentrations offolate. These samples were tested against an appropriate control and the observed bias ispresented in the following table:

SubstanceSubstance Test Concentrationmg/dL

Analyte Concentrationng/mL (nmol/L) Bias (%)

Acetaminophen 200 3.7 (8.4) -0.2200 8.6 (19.5) -0.7

Acetylsalicylic Acid 1000 3.4 (7.7) -0.71000 8.2 (18.6) 9.6

Ibuprofen 50 3.8 (8.6) -5.050 9.6 (21.7) 0.0

Acetylcysteine 566 3.4 (7.7) 5.3566 9.4 (21.3) 3.8

Ampicillin-Na 1000 3.4 (7.7) -0.71000 9.1 (20.6) -0.6

Cefoxitin 660 3.2 (7.2) -2.9660 9.5 (21.5) -7.1

Cyclosporine 5 3.1 (7.0) 1.55 7.8 (17.7) -3.0

Doxycyclin 50 3.0 (6.8) 5.550 7.0 (15.9) 8.7


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SubstanceSubstance Test Concentrationmg/dL


Levodopa 20 3.1 (7.0) 2.020 8.5 (19.3) -1.9

Methyldopa 20 3.7 (8.4) 3.720 10.4 (23.6) 0.7

Metronidazole 200 3.7 (8.4) -1.0200 11.1 (25.1) -9.8

Phenylbutazone 40 3.6 (8.2) 1.440 9.4 (21.3) 0.9

Rifampicin 60 3.6 (8.2) -9.360 10.5 (23.8) -7.9

Theophylline 10 3.7 (8.4) -3.510 9.9 (22.4) -0.9

Ascorbic Acid 44 3.4 (7.7) 6.644 10.0 (22.7) 6.1

The Atellica IM Fol assay is designed to have ≤ 10% interference for bilirubin and lipemia whentested at the levels indicated in the table below.Bias is the difference in the results between the control sample (does not contain theinterferent) and the test sample (contains the interferent) expressed in percent. Analyteresults should not be corrected based on this bias.

SubstanceSubstance Test ConcentrationCommon Units (SI Units)


Bilirubin, conjugated 20 mg/dL (341 µmol/L) 3.83 (8.67) 0.520 mg/dL (341 µmol/L) 9.50 (21.52) 9.2

Bilirubin, unconjugated 20 mg/dL (341 µmol/L) 3.62 (8.20) -5.220 mg/dL (341 µmol/L) 9.80 (22.20) -1.4

Lipemia (Intralipid®) 2000 mg/dL (22.7 mmol/L) 5.02 (11.37) 1.82000 mg/dL (22.7 mmol/L) 8.79 (19.91) 3.0

Biotin was added to serum samples containing different concentrations of folate. Thesesamples were tested against an appropriate control and the observed bias is presented in thefollowing table:


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AnalyteConcentrationng/mL (nmol/L)

Biotin Test Level in Serum(ng/mL)

0 1 5 19 38 75 150 300 600% Bias

9.23 (20.91) — 1.4 -1.1 6.9 4.7 20.2 > MIa > MI > MI

17.37 (39.34) — 1.3 1.3 3.6 8.3 35.3 > MI > MI > MIa Measuring Interval

In addition, biotin was added to whole blood samples containing different concentrations offolate. These samples were tested against an appropriate control and the observed bias ispresented in the following table:

AnalyteConcentrationng/mL (nmol/L)

Biotin Test Level in Whole Blood(ng/mL)

0 1 5 19 38 75 150 300 600% Bias

12.76 (28.90) — -7.0 -8.4 -8.8 -5.7 -3.5 -1.1 49.6 68.018.47 (41.83) — 0.1 4.5 -0.2 4.4 7.8 12.7 > MIa > MI

a Measuring IntervalSpecimens that contain biotin at a concentration of 50 ng/mL (in serum) or 75 ng/mL (inwhole blood) demonstrate a less than or equal to 10% change in results. Biotin concentrationsgreater than these may lead to falsely elevated results for patient samples.The recommended adult daily dietary intake for biotin is 30 µg/day. Over the counter dietarysupplements promoted for use in hair, skin, and nail health may contain 5–100 mg of biotin,with recommendations to take multiple pills per day. Pharmacokinetic studies in healthy adultshave shown that, in subjects ingesting 5 mg, 10 mg, and 20 mg of biotin, serumconcentrations of biotin can reach up to 73 ng/mL, 141 ng/mL, and 355 ng/mL, respectively.16

Subjects who take up to 300 mg of biotin per day may have plasma biotin levels as high as1160 ng/mL.17

Dilution RecoveryFive human serum samples in the range of 18.93–32.42 ng/mL (42.88–73.43 nmol/L) offolate were diluted 1:2 with Atellica IM Fol DIL and assayed for recovery and parallelism. Therecoveries ranged from 102.6%–110.0% with a mean of 105.4%.

Sample DilutionObserved(ng/mL)

Expected(ng/mL)

Observed(nmol/L)

Expected(nmol/L)

Recovery(%)

1 1:2 33.08 31.15 74.93 70.55 106.22 1:2 28.46 27.27 64.46 61.77 104.43 1:2 19.42 18.93 43.99 42.88 102.64 1:2 24.44 22.21 55.36 50.31 110.05 1:2 33.66 32.42 76.24 73.43 103.8Mean 105.4


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Results were established using the Atellica IM Analyzer. Assay results obtained at individuallaboratories may vary from the data presented.

Spiking RecoveryVarying amounts of folate were added to 5 samples with endogenous folate levels of1.05–1.35 ng/mL (2.38–3.06 nmol/L). The recoveries ranged from 87%–116% with a mean of104%.

SampleObserved(ng/mL)

Expected(ng/mL)

Observed(nmol/L)

Expected(nmol/L)

Recovery(%)

1 4.65 4.32 10.53 9.79 1088.50 7.58 19.24 17.16 11212.21 10.62 27.66 24.04 11516.43 15.11 37.21 34.22 10918.32 16.90 41.50 38.27 108

Mean 110

2 4.65 4.45 10.54 10.08 1048.40 7.60 19.03 17.22 11112.27 10.59 27.79 23.99 11615.49 15.39 35.09 34.86 10119.61 16.93 44.42 38.36 116

Mean 110

3 4.45 4.70 10.09 10.65 957.80 7.74 17.68 17.53 10111.45 10.80 25.93 24.46 10613.60 15.71 30.79 35.59 8717.05 17.18 38.62 38.90 99

Mean 98

4 3.97 4.21 8.99 9.54 946.74 6.79 15.28 15.37 999.40 9.46 21.29 21.42 9911.64 11.72 26.35 26.54 9914.52 14.06 32.90 31.85 103

Mean 99

5 4.11 4.06 9.30 9.20 1017.21 6.79 16.33 15.37 10610.08 9.36 22.84 21.21 10811.90 11.86 26.95 26.86 100


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SampleObserved(ng/mL)

Expected(ng/mL)

Observed(nmol/L)

Expected(nmol/L)

Recovery(%)

14.86 14.13 33.67 32.01 105Mean 104

Mean 104

Results were established using the Atellica IM Analyzer. Assay results obtained at individuallaboratories may vary from the data presented.

StandardizationThe Atellica IM Fol assay standardization is traceable to an internal standard manufacturedusing highly purified material (N‑5‑methyl tetrahydrofolate). Assigned values for calibrators aretraceable to this standardization.

Technical AssistanceFor customer support, contact your local technical support provider or distributor.siemens.com/healthineers

References1. Miale JB. Laboratory Medicine: Hematology. 6th ed. St. Louis: CV Mosby; 1982:416–440.2. Brewster MA. Vitamins. In: Kaplan LA, Pesce AJ, eds. Clinical Chemistry: Theory, Analysis,

Correlation. St. Louis: CV Mosby; 1989:543–568.3. Steinkamp RC. Vitamin B12 and folic acid: clinical and pathophysiological considerations.

In: Brewster MA, Naito HK, eds. Nutritional Elements and Clinical Biochemistry. New York:Plenum Publishing Corp.; 1980:169–240.

4. McNeely MD. Folic acid. In: Pesce AJ, Kaplan LA, eds. Methods in Clinical Chemistry. St.Louis: CV Mosby; 1987:539–542.

5. Centers for Disease Control. Perspectives in disease prevention and health promotionupdate: Universal precautions for prevention of transmission of human immunodeficiencyvirus, hepatitis B virus and other bloodborne pathogens in healthcare settings. MMWR.1988;37(24):377–382, 387–388.

6. Clinical and Laboratory Standards Institute. Procedures for the Handling and Processing ofBlood Specimens for Common Laboratory Tests; Approved Guideline—Fourth Edition.Wayne, PA: Clinical and Laboratory Standards Institute; 2010. CLSI Document GP44‑A4.

7. Clinical and Laboratory Standards Institute. Protection of Laboratory Workers FromOccupationally Acquired Infections; Approved Guideline—Fourth Edition. Wayne, PA:Clinical and Laboratory Standards Institute; 2014. CLSI Document M29‑A4.

8. Mastropaolo W, Wilson M. Effect of light on serum B12 and folate stability. Clin Chem.1993;39(5):913.

9. Clinical and Laboratory Standards Institute. Procedures for the Collection of DiagnosticBlood Specimens by Venipuncture; Approved Standard—Sixth Edition. Wayne, PA: Clinicaland Laboratory Standards Institute; 2007. CLSI Document GP41‑A6.

10. Clinical and Laboratory Standards Institute. Tubes and Additives for Venous and CapillaryBlood Specimen Collection; Approved Standard—Sixth Edition. Wayne, PA: Clinical andLaboratory Standards Institute; 2010. CLSI Document GP39‑A6.


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11. Clinical and Laboratory Standards Institute. Defining, Establishing, and Verifying ReferenceIntervals in the Clinical Laboratory; Approved Guideline—Third Edition. Wayne, PA:Clinical and Laboratory Standards Institute; 2010. CLSI Document EP28‑A3c.

12. Clinical and Laboratory Standards Institute. Evaluation of Detection Capability for ClinicalLaboratory Measurement Procedures; Approved Guideline—Second Edition. Wayne, PA:Clinical and Laboratory Standards Institute; 2012. CLSI Document EP17‑A2.

13. Clinical and Laboratory Standards Institute. Evaluation of Precision of QuantitativeMeasurement Procedures; Approved Guideline—Third Edition. Wayne, PA: Clinical andLaboratory Standards Institute; 2014. CLSI Document EP05‑A3.

14. Clinical and Laboratory Standards Institute. Measurement Procedure Comparison and BiasEstimation Using Patient Samples; Approved Guideline—Third Edition. Wayne, PA: Clinicaland Laboratory Standards Institute; 2013. CLSI Document EP09‑A3.

15. Clinical and Laboratory Standards Institute. Interference Testing in Clinical Chemistry;Approved Guideline—Second Edition. Wayne, PA: Clinical and Laboratory StandardsInstitute; 2005. CLSI Document EP07‑A2.

16. Grimsey P, Frey N, Bendig G, et al. Population pharmacokinetics of exogenous biotin andthe relationship between biotin serum levels and in vitro immunoassay interference. Int. J.Pharmacokinet. 2017;2(4):247–256.

17. Piketty ML, Prie D, Sedel F, et al. High-dose biotin therapy leading to false biochemicalendocrine profiles: validation of a simple method to overcome biotin interference. ClinChem Lab Med. 2017;55(6):817‑825.

Definition of SymbolsThe following symbols may appear on the product labeling:

Symbol Symbol Title and DescriptionConsult instructions for use

Version of instructions for use

Internet URL address to access the electronic instructions for use

Revision

CautionConsult instructions for use or accompanying documents for cautionary informationsuch as warnings and precautions that cannot, for a variety of reasons, be presentedon the medical device.Biological risksPotential biological risks are associated with the medical device.

Corrosive

Dangerous to environment


11200602_EN Rev. 04, 2020-11 21 / 24

Symbol Symbol Title and DescriptionIrritantOral, dermal, or inhalation hazard

Inhalation hazardRespiratory or internal health

FlammableFlammable to extremely flammable

Oxidizing

Explosive

Toxic

Compressed gas

Keep away from sunlightPrevent exposure to sunlight and heat.

UpStore in an upright position.

Do not freeze

Temperature limitUpper and lower limits of temperature indicators are adjacent to the upper andlower horizontal lines.Handheld barcode scanner

In vitro diagnostic medical device

Contains sufficient for <n> testsTotal number of IVD tests the system can perform with the IVD kit reagents appearsadjacent to the symbol.Prescription device (US only)Applies only to United States-registered IVD assays.CAUTION: Federal (USA) law restricts this device to sale by or on the order of alicensed healthcare professional.Mixing of substancesMix product before use.


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Symbol Symbol Title and DescriptionReconstitute and mix lyophilized product before use.

Target

Interval

Legal Manufacturer

Authorized Representative in the European Community

Use-by dateUse by the designated date.

Batch code

Catalog number

Recycle

Printed with soy ink

CE Mark

CE Mark with notified body ID numberNotified body ID number can vary.

Date format (year‑month‑day)Variable hexadecimal number that ensures the Master Curve and Calibratordefinition values entered are valid.Common Units

International System of Units

Material

Unique material identification number

Name of control

Type of control


11200602_EN Rev. 04, 2020-11 23 / 24

Legal InformationAtellica, ReadyPack, and ADVIA Centaur are trademarks of Siemens Healthcare Diagnostics.All other trademarks and brands are the property of their respective owners.© 2017–2020 Siemens Healthcare Diagnostics. All rights reserved.

Siemens Healthcare Diagnostics Inc.511 Benedict AvenueTarrytown, NY 10591USAsiemens.com/healthineers Siemens Healthineers HeadquartersSiemens Healthcare GmbHHenkestr. 12791052 ErlangenGermanyPhone: +49 9131 84-0siemens.com/healthineers


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Documents

Folate (Fol) Atellica IM Analyzer