AnemoCheck
Device Facts
| Record ID | K163215 |
|---|---|
| Device Name | AnemoCheck |
| Applicant | Sanguina, LLC |
| Product Code | KHG · Hematology |
| Decision Date | Sep 13, 2017 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 864.7500 |
| Device Class | Class 2 |
| Attributes | Pediatric |
Indications for Use
The AnemoCheck is an in vitro diagnostic semi-quantitative assay for use in point-of-care, clinical and doctor office laboratories for the determination of hemoglobin level and estimation of hematocrit percentage (within normal hemoglobin range) in anticoagulated (K2EDTA, heparin or citrate) whole blood (capillary or venous). AnemoCheck should not be used to evaluate neonatal samples (birth – 1 month).
Device Story
AnemoCheck is a manual, semi-quantitative colorimetric assay for total hemoglobin determination and hematocrit estimation. Input: anticoagulated whole blood (capillary or venous). Operation: manual test requiring no electrical power or additional equipment; 2-minute development time. Output: visual colorimetric result indicating hemoglobin level/hematocrit percentage. Used in point-of-care, clinical, and doctor office settings by healthcare professionals. Results provide rapid hemoglobin assessment to assist clinical decision-making regarding patient anemia status.
Clinical Evidence
Method comparison study performed at three point-of-care sites with 262 patient samples (221 venous, 41 capillary) across a diverse population (pediatric to 92 years old) including various anemias. Comparison against HemoCue Hb 201+ showed linear regression R-values of 0.979 (venous) and 0.972 (capillary). Analytical performance included precision (repeatability and reproducibility), linearity (4.0–18.0 g/dL), and interference testing. No clinical diagnostic sensitivity/specificity studies were required as the device is a semi-quantitative screening tool.
Technological Characteristics
Semi-quantitative colorimetric assay. Manual test; no electrical power or additional equipment required. Compatible with K2EDTA, heparin, or citrate anticoagulated whole blood. 2-minute development time.
Indications for Use
Indicated for determination of hemoglobin level and estimation of hematocrit in patients >1 month of age using capillary or venous whole blood. Contraindicated for neonatal samples (birth – 1 month).
Regulatory Classification
Identification
A whole blood hemoglobin assay is a device consisting or reagents, calibrators, controls, or photometric or spectrophotometric instrumentation used to measure the hemoglobin content of whole blood for the detection of anemia. This generic device category does not include automated hemoglobin systems.
Special Controls
*Classification.* Class II (special controls). An acid hematin intended for use with whole blood hemoglobin assays is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 864.9.
Predicate Devices
Related Devices
- K221508 — AnemoCheck Home · Sanguina, Inc. · Sep 29, 2023
- K122553 — MISSION PLUS HB HEMOGLOBIN TESTING SYSTEM · ACON Laboratories, Inc. · Aug 5, 2013
Submission Summary (Full Text)
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY
A. 510(k) Number:
K163215
B. Purpose for Submission:
Clearance of a new device
C. Measurand:
Hemoglobin
D. Type of Test:
Semi-quantitative, colorimetric measurement of hemoglobin and estimation of hematocrit
E. Applicant:
Sanguina, LLC
F. Proprietary and Established Names:
Proprietary Name – AnemoCheck™
Common Name – AnemoCheck™ Whole Blood Hemoglobin Determination
G. Regulatory Information:
1. Regulation section:
CFR 864.7500
2. Classification:
Class II
3. Product code:
KHG - Whole blood hemoglobin assays
4. Panel:
(81) Hematology
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H. Intended Use:
1. Intended use(s):
The AnemoCheck is an in vitro diagnostic semi-quantitative assay for use in point-of-care, clinical and doctor office laboratories for the determination of hemoglobin level and estimation of hematocrit percentage (within normal hemoglobin range) in anticoagulated (K2EDTA, heparin or citrate) whole blood (capillary or venous). AnemoCheck should not be used to evaluate neonatal samples (birth – 1 month).
2. Indication(s) for use:
Same as Intended Use
3. Special conditions for use statement(s):
For Prescription Use Only
Results outside of the AnemoCheck reportable range must be confirmed by a clinical laboratory and/or medical history.
AnemoCheck results should be confirmed by a quantitative test when monitoring disease progression or making treatment decisions.
4. Special instrument requirements:
Not applicable
I. Device Description:
The AnemoCheck™ is a semi-quantitative colorimetric assay for determination of total hemoglobin (g/dL) and estimated hematocrit (%) within the normal range from whole blood. This device can be used with capillary or venous whole blood samples. The assay is a rapid (development time is 2 minutes) manual test that does not require electrical power or additional equipment. The test is designed to use a plastic 5 μL capillary tube coated with sodium heparin (for anticoagulation) and 0.5 mL chemical reagent solution pre-filled in a 1.8 mL borosilicate glass vial. The capillary tube serves as a pipette and measuring instrument. Sample dilution is not required.
J. Substantial Equivalence Information:
1. Predicate device name(s):
HemoCue® Hb 201+ System
2. Predicate 510(k) number(s):
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K032203
3. Comparison with predicate:
| Similarities | | |
| --- | --- | --- |
| Item | AnemoCheckTM | HemoCue® Hb 201+ System (K032203) |
| Method | Colorimetric oxidation reduction chemical reaction with hemoglobin | Same |
| Storage Conditions | 15–30°C | Same |
| Single Use | Yes | Same |
| Setting | Point-of-Care, Doctor's office and Clinical Laboratories | Same |
| Differences | | |
| --- | --- | --- |
| Item | AnemoCheckTM | HemoCue® Hb 201+ System (K032203) |
| Intended Use | The AnemoCheck is an in vitro diagnostic semi-quantitative assay for use in point-of-care, clinical and doctor office laboratories for the determination of hemoglobin level and estimation of hematocrit percentage (within normal hemoglobin range) in anticoagulated (K2EDTA, heparin or citrate) whole blood (capillary or venous). AnemoCheck should not be used to evaluate neonatal samples (birth – 1 month). | Quantitative determination of hemoglobin in capillary, venous and arterial whole blood. The HemoCue® Hb 201 Microcuvettes are intended for use in the HemoCue® Hb 201+ analyzer. The HemoCue 201 system is used for the quantitative determination of hemoglobin in blood using a specially designed analyzer, the HemoCue Hb 201+ analyzer, and specially designed microcuvettes the HemoCue Hb 201 Microcuvettes. The HemoCue Hb 201+ analyzer is only to be used with HemoCue Hb 201 Microcuvettes. The reagents/microcuvettes and the analyzer form an analytical system. The HemoCue Hb 201+ analyzer is only to be used together with HemoCue Hb 201 microcuvettes. The use of any other device in the HemoCue Hb 201+ analyzer except the HemoCue Hb 201 microcuvettes is neither supported nor recommended by HemoCue, and could give erroneous results with serious clinical consequences. The HemoCue Hb 201 microcuvettes are for in vitro |
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| Differences | | |
| --- | --- | --- |
| Item | AnemoCheck™ | HemoCue® Hb 201^{®} System (K032203) |
| | | diagnostic use only. |
| Analyte | Hemoglobin (g/dL) and estimated hematocrit percentage (%) with normal range | Hemoglobin (g/dL) |
| Result | Semi-quantitative | Quantitative |
| Read out | Manual (visual-based) | Automated (reader-based) |
| Development Time | 2 minutes | ~1 minute (time to read) |
| Quality Control/Calibration | Streck Inc. High Whole Blood Hemoglobin Control | Hemotrol Low, Mid and High Whole Blood Hemoglobin Controls |
| Anticoagulant(s) | K_{2}EDTA, citrate or heparin | EDTA or heparin |
| Specimen Type | Capillary and venous whole blood | Capillary, arterial and venous whole blood |
| Packaging | Disposable Test Vial | Disposable Cuvette |
| Shelf-Life | 9 months | 24 months |
| Sample Volume | 10 μL from fingerstick and 20 μL from venous tube | 10 μL |
## K. Standard/Guidance Document Referenced (if applicable):
ISO 14971 Second Edition 2007-03-01, Medical devices - Application of Risk Management to Medical Devices
CLSI EP09-A3; Measurement Procedure Comparison and Bias Estimation Using Patient Samples; Approved Guideline - Third Edition
CLSI EP05-A3; Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline – Third Edition
CLSI EP06-A; Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline
CLSI EP25-A; Evaluation of Stability of In Vitro Diagnostic Reagents; Approved Guideline
## L. Test Principle:
The reaction in each AnemoCheck test vial is an oxidation-reduction reaction utilizing 3,3'-5,5 tetramethylbenzidine (TMB). Erythrocyte membranes are disintegrated via high osmotic pressure when exposed to a chemical solution, releasing the hemoglobin. TMB is then oxidized in the presence of hemoglobin and hydrogen peroxide within the solution, exhibiting stable color products that are discernible by visual interpretation.
To determine hemoglobin values from this test, 5 μL of the capillary tube is filled with blood
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(via capillary action or "wicking") and then delivered to the AnemoCheck test vial which is pre-filled with the TMB chemical solution. After mixing the blood with solution and 2 minutes of development, the reaction with whole blood is complete. The resulting color of the solution then allows for visual interpretation with the color card for determination of hemoglobin (g/dL) and estimated hematocrit percentage (%).
## M. Performance Characteristics (if/when applicable):
### 1. Analytical performance:
#### a. Precision/Reproducibility:
Precision studies were conducted to assess the sources of potential variation for the AnemoCheck test, namely variance due to site, operator, and lot.
#### Repeatability
Repeatability studies were conducted at a point-of-care site using three different AnemoCheck test lots and three operators. Five natural patient samples collected in sodium citrate tubes with hemoglobin concentrations at the lower and upper limits and medical decision levels of the analytical measuring range of the AnemoCheck were tested. Each sample was tested 10 times for a total of 90 tests results for each hemoglobin concentration. Repeatability results were within the defined acceptance criteria.
| Sample | N | Mean (g/dL) | Within -Run | | Between-Lot | | Between-Operator | | Total | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| 630P7 | 90 | 4.8 | 0.16 | 3.36 | 0.08 | 1.70 | 0.15 | 3.21 | 0.27 | 5.63 |
| 630P5 | 90 | 14.2 | 0.21 | 1.46 | 0.10 | 0.71 | 0.20 | 1.39 | 0.48 | 3.36 |
| 706P19 | 90 | 14.8 | 0.20 | 1.36 | 0.13 | 0.86 | 0.17 | 1.13 | 0.34 | 2.30 |
| 706P14 | 90 | 6.4 | 0.24 | 3.73 | 0.11 | 1.70 | 0.23 | 3.57 | 0.48 | 7.49 |
| 706P12 | 90 | 17.7 | 0.20 | 1.14 | 0.11 | 0.62 | 0.17 | 0.96 | 0.39 | 2.19 |
### Reproducibility
Reproducibility was conducted at three point-of-care sites over five different days using three different lots of AnemoCheck tests at each site and one lot of Streck Para 4 controls (K842017, low and high levels) across all sites. Testing was performed once daily using the same set of controls for 5 days. Each control level was run 10 times by two operators at each site. A total of 300 test results were generated for each control level at each site. SD and %CV for within-run, between-day, between-operator, and between-site were calculated for each site and all sites combined. Results for each site and all sites combined were within the predefined acceptance criteria.
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All Sites Combined
| Sample | N | Mean | Within-Run | | Between-Day | | Between-Lot | | Between-Operator | | Between-Site | | Total | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Low | 900 | 5.7 | 0.26 | 4.54 | 0.10 | 1.80 | 0.12 | 2.13 | 0.21 | 3.64 | 0.10 | 1.68 | 0.36 | 6.40 |
| High | 900 | 17.3 | 0.33 | 1.92 | 0.21 | 1.22 | 0.09 | 0.51 | 0.15 | 0.89 | 0.03 | 0.20 | 0.54 | 3.11 |
Site 1
| Sample | N | Mean | Within-Run | | Between-Day | | Between-Lot | | Between-Operator | | Total | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Low | 300 | 5.6 | 0.31 | 5.50 | 0.10 | 1.76 | 0.14 | 2.43 | 0.33 | 5.88 | 0.42 | 7.48 |
| High | 300 | 17.3 | 0.37 | 2.13 | 0.28 | 1.64 | 0.15 | 0.87 | 0.22 | 1.28 | 0.49 | 2.87 |
Site 2
| Sample | N | Mean | Within-Run | | Between-Day | | Between-Lot | | Between-Operator | | Total | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Low | 300 | 5.8 | 0.22 | 3.79 | 0.17 | 2.92 | 0.10 | 1.66 | 0.06 | 1.11 | 0.30 | 5.19 |
| High | 300 | 17.3 | 0.41 | 2.39 | 0.32 | 1.85 | 0.09 | 0.49 | 0.23 | 1.32 | 0.72 | 4.16 |
Site 3
| Sample | N | Mean | Within-Run | | Between-Day | | Between-Lot | | Between-Operator | | Total | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Low | 300 | 5.7 | 0.21 | 3.64 | 0.12 | 2.14 | 0.15 | 2.61 | 0.06 | 0.98 | 0.34 | 5.96 |
| High | 300 | 17.3 | 0.17 | 0.98 | 0.08 | 0.46 | 0.07 | 0.40 | 0.07 | 0.41 | 0.32 | 1.87 |
b. Linearity/assay reportable range:
Two linearity studies were conducted for AnemoCheck. The first linearity study was conducted using 13 natural patient samples (5.0–17.7 g/dL) correlating to AnemoCheck color blocks on the test card. Samples were tested in triplicate using three lots of AnemoCheck tests. Color change in the AnemoCheck test vials were interpreted visually and recorded. The second linearity study utilized contrived samples across the reportable range (4.0–18.0 g/dL). Samples were tested in triplicate using three lots of AnemoCheck tests. Results were interpreted visually and by spectrophotometry. Linear regression of the visual results was performed and based on the data analysis, AnemoCheck demonstrated linearity over the claimed measuring range of 4.0–18.0 g/dL.
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
Open-vial and Closed-vial Stability
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Open-vial stability was determined by testing natural patient samples across the reportable range using three lots AnemoCheck tests. Each sample was tested in triplicate. Under room temperature conditions (15–30°C), samples were tested immediately after opening the vial containing the AnemoCheck chemical reagent solution (t=0) and at 15 minutes. The acceptance criterion for open-vial stability was met at 15 minutes. The open-vial stability study supports a claim for 10 minutes after opening when stored at 15–30°C.
Closed-vial stability was determined by testing control material across the reportable range using three lots AnemoCheck tests. Each sample was tested in triplicate. Samples were tested immediately after opening the AnemoCheck chemical reagent solution vial (t=0), 156 days, and 280 days at 15–30°C. The closed-vial stability study supports a claim for 9 months when stored at 15–30°C.
## Transport Stability
Three different AnemoCheck test lots were tested after being exposed to different transport conditions which included vibration testing for various time frames (8 hours, 1 day and 5 days), extreme temperature (45–50°C, 60°C and -20°C) and prolonged humidity (7 days). AnemoCheck tests were from lots that were stored for 260, 262 and 264 days after being manufactured. Patient whole blood samples spanning medical decision points and a low control (4.0, 10.0, 13–15.0 and 18.0 g/dL) were used to evaluate the AnemoCheck test kits exposed to the different transport conditions. Acceptance criteria were met for all conditions with the exception of studies conducted for the test stored at -20°C for 5 days. The labeling adequately addresses the limitation that AnemoCheck tests should not be stored frozen.
## Value Assignment
Value assignment of control material (Streck High hemoglobin control, K842017) was conducted with two operators using two lots of control material on two different days with two different lots of AnemoCheck for a total of 67 test results. The operator was instructed to fill an AnemoCheck capillary tube halfway with a control sample in order to obtain 6.0 g/dL Hgb value. AnemoCheck test results were quantified (via spectrophotometer) to enable direct comparison of hemoglobin values to the expected hemoglobin control result of 6.0 g/dL. The results confirmed that 6.0 g/dL Hgb value corresponds to AnemoCheck capillary tube filled halfway.
## Quality Controls
Quality control testing is recommended to be performed prior to use. A hemoglobin test capturing a hemoglobin level of approximately 6.0 g/dL is utilized to ensure the redox reaction color change can proceed. The user is instructed to fill an AnemoCheck capillary tube halfway with a control sample (Streck, Inc. High hemoglobin control, K842017) and test it using an AnemoCheck test vial. After
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mixing and development time (2 minutes), the resulting color should correlate to approximately $6\mathrm{g / dL}$ on the AnemoCheck color scale.
# d. Detection limit:
In order to determine the detection limit (LoD) of the AnemoCheck assay (4.0–18.0 g/dL), four low hemoglobin samples 0.0, 0.2, 0.5 and $1.4\mathrm{g / dL}$ as verified by the predicate device were used. Each sample was tested 10 times using one lot of AnemoCheck test. Testing was completed as an end-user would perform testing per the instructions for use. The blank $0.0\mathrm{g / dL}$ did not elicit any visual color change and all other samples elicited a blue result correlating to the lowest color block in the reportable range when interpreted between 2–10 minute development time. The limit of detection (LoD) is $0.2\mathrm{g / dL}$ .
# e. Analytical specificity:
Studies were conducted where the interfering substance was introduced into a subject, and then compared against control samples drawn from the same subject. Venous blood samples containing ibuprofen, acetaminophen, aspirin, Benadryl, bilirubin and triglycerides with the maximum concentrations listed below were tested in triplicate using one lot of AnemoCheck.
In order to assess potential interference with blood pH and extremely high bilirubin and albumin concentrations, studies were conducted by delivering a concentrated dose of a potentially interfering substance ("spiking") to healthy venous blood samples of known volume in order to reach final concentrations. Hemoglobin concentrations were verified on a reference standard hematology analyzer that covered the analytical measuring range (4.0, 10, 13.0–15.0, 18.0 g/dL). Samples were then tested in triplicate using one lot of AnemoCheck. Potential interfering substances at the concentration in the table below and blood pH values between 6.8 and 8.4 do not interfere with the AnemoCheck test.
Interfering Substance Concentrations
| Interfering Substance | Maximum Concentration Tested |
| --- | --- |
| Acetaminophen | 1000 mg |
| Aspirin | 800 mg |
| Diphenhydramine HCl (Benadryl) | 50 mg |
| Bilirubin conjugated with albumin | 20 mg/dL |
| Ibuprofen | 800 mg |
| Triglycerides | 350 mg/dL |
| Blood pH | pH 6.8 |
| Alkaline Blood pH | pH 8.4 |
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f. Assay cut-off:
# 2. Comparison studies:
a. Method comparison with predicate device:
Method comparison studies were performed at three point-of-care clinical sites in the United States to assess the performance of AnemoCheck compared to the predicate device (HemoCue Hb 201+ System). A total of 262 patient samples, which included 221 venous samples and 41 capillary samples, were used to assess the AnemoCheck test. Of the 221 venous samples, 140 samples that were collected in $\mathrm{K}_2\mathrm{EDTA}$ tubes, 34 samples were collected in citrate tubes and 47 samples were collected in heparin tubes. Of the capillary samples, 30 samples were collected in $\mathrm{K}_2\mathrm{EDTA}$ tubes and 11 samples were collected in heparin tubes. In total 170, 34 and 58 patient samples were collected in $\mathrm{K}_2\mathrm{EDTA}$ , citrate tubes and heparin tubes, respectively. Fourteen samples of the 262 patient samples used in this study were contrived. Testing was performed using three different lots of AnemoCheck (one lot at each site) with five operators over three sites. The study population consisted of $48.1\%$ males and $51.9\%$ females including pediatric (i.e. infants, children, and adolescents) and adult subjects up to 92 years of age. In addition to healthy individuals, subjects with the following clinical conditions were included in the studies: microcytic hypochromic anemia, thalassemia, sickle cell anemia, acute lymphoblastic leukemia (ALL), aplastic anemia, acute idiopathic thrombocytopenic purpura (ITP), and iron deficiency anemia. Linear regression analyses demonstrate comparable performance between the AnemoCheck and HemoCue Hb 201+ System across the analytical measuring range. The method comparison study demonstrated that the analytical performance of the AnemoCheck test is substantially equivalent to the predicate device.
| Sample type | N | Min Hgb (g/dL) | Max Hgb (g/dL) | Regression | R value |
| --- | --- | --- | --- | --- | --- |
| Venous | 221 | 4.0 | 17.5 | y = 1.009x - 0.131 | 0.979 |
| Capillary | 41 | 6.5 | 17.0 | y = 0.9452x + 0.4932 | 0.972 |
| Anticoagulant Type | N | Min Hgb (g/dL) | Max Hgb (g/dL) | Regression | R Value |
| --- | --- | --- | --- | --- | --- |
| Citrate | 34 | 5.0 | 17.0 | y = 1.01x + 0.016 | 0.98 |
| Heparin | 58 | 4.0 | 17.0 | y = 0.97x + 0.172 | 0.97 |
| K2EDTA | 170 | 4.0 | 18.0 | y = 1.02x - 0.224 | 0.97 |
b. Matrix comparison:
All matrices are incorporated in the method comparison study.
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3. Clinical studies:
a. Clinical Sensitivity:
Not applicable
b. Clinical specificity:
Not applicable
c. Other clinical supportive data (when a. and b. are not applicable):
4. Clinical cut-off:
Not applicable
5. Expected values/Reference range
The reference range was verified along with the 95% CI using 85 normal adult samples and 74 normal pediatric samples using three lots of AnemoCheck tests as outlined in the table below. The reference range intervals provided were further supported by published literature¹.
| Subgroups | Average Hgb Value (g/dL) | Lower 95% CI (g/dL) | Upper 95% CI (g/dL) | Expected Values (g/dL) |
| --- | --- | --- | --- | --- |
| Infant (>1 month – 2y) | 11.7 | 11.3 | 12.1 | 11.0–14.0 |
| Child (3–12y) | 11.2 | 10.5 | 11.8 | 11.0–14.0 |
| Adolescent (13–21y) | 14.2 | 13.8 | 14.6 | 11.0–14.0 |
| Adult Male >22y | 15.2 | 14.6 | 15.8 | 13.0–17.0 |
| Adult Female >22y | 12.4 | 11.9 | 12.9 | 12.0–15.0 |
N. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.
O. Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
¹ WHO. Haemoglobin concentrations for the diagnosis of anaemia and assessment of severity. Vitamin and Mineral Nutrition Information System. Geneva, World Health Organization, 2011 (WHO/NMH/NHD/MNM/11.1).
