The Astoria-Pacific SPOTCHECK G6PD 50 Hour Reagent Kit is intended for the semi-quantitative determination of glucose-6-phosphate dehydrogenase activity in erythrocytes using the Astoria-Pacific SPOTCHECK Analyzer. Measurements of glucose-6-phosphate dehydrogenase activity are used primarily in the diagnosis and treatment of nonspherocytic congenital hemolytic anemia or drug-induced hemolytic anemia associated with a G6PD deficiency. It is for in vitro diagnostic use primarily as an aid in screening for decreased levels of G6PD enzyme activity. This device is for use by trained, qualified laboratory personnel.

Device Description

The Astoria-Pacific SPOTCHECK G6PD 50 Hour Reagent Kit is a reagent kit used for the semi-quantitative determination of glucose-6-phosphate dehydrogenase activity in erythrocytes. The method is based on the spectrophotometric method where enzyme activity is measured by observing NADP+ reduction to NADPH when glucose-6-phosphate is present as a substrate. Maleimide is added to inhibit the production of NADPH by 6-phosphogluconate dehydrogenase. The fluorescent NADPH produced is proportional to the G6PD enzyme activity. The kit contains Extraction Buffer, Tris Buffer, G6PD Substrate, and NADH Stock Std reagents.

AI/ML Overview

The provided text describes the G6PD 50-Hour Reagent Kit and its performance characteristics. Here's a breakdown of the requested information:

1. Table of Acceptance Criteria and Reported Device Performance

The document doesn't explicitly state formal "acceptance criteria" for regulatory approval in the typical sense of numerical thresholds for sensitivity, specificity, etc., that the device had to meet to be cleared. Instead, it presents performance characteristics that demonstrate the device is substantially equivalent to a predicate. The key performance aspects highlighted are:

Performance Characteristic	Acceptance Criteria (Implicit)	Reported Device Performance
Intended Use	Semi-quantitative determination of G6PD activity for screening decreased levels in erythrocytes, aiding diagnosis/treatment of G6PD deficiency.	Semi-quantitative determination of G6PD activity in erythrocytes for screening decreased levels of G6PD enzyme activity.
Chemical Principle	NADP+ Reduction to NADPH.	NADP+ Reduction to NADPH.
Temperature	Operation at a specific temperature.	37° C.
Stability	Reagent stability.	8 hours.
Expected Value Range	Ability to delineate normal, intermediate, and deficient G6PD activity ranges in the tested populations.	Normal Population: 40 - 224 µM NADPH (Average 150 µM, Range 42-224 µM). Deficient Population: 4 - 40 µM NADPH (Average 18 µM, Range 4-40 µM). Intermediate Population: 28 - 69 µM NADPH (Average 49 µM, Range 28-69 µM).
Sensitivity	Ability to discern low levels of NADPH (reflecting G6PD activity).	As little as 2 µM NADPH is discernible from no response.
Detection Limit	Lower limit of quantifiable activity.	9 µM NADPH.
Detection Method	Method of detecting NADPH.	Fluorescence of NADPH (λ excit. = 350 nm, λ emiss. = 450 nm).
Linearity	Correlation between response and known NADPH concentrations.	Response standards from 0 to 75 µM NADH gave a correlation coefficient r > .999.
Carryover	Minimal sample-to-sample interference.	Less than 2%; corrected by CARRYOVER cup.
Specificity	Specific for G6PD activity.	Achieved by using specific substrate (glucose-6-phosphate). Small NADPH production in absence of substrate.
Within-run Precision	Consistency of results within a single run.	G6PD Deficient: CV 3.2%. Intermediate: CV 4.4%. G6PD Normal: CV 3.1%.
Total Precision	Overall consistency of results across runs and days.	G6PD Deficient: CV 5.4%. Intermediate: CV 6.0%. (Normal not provided for total precision but within-run is similar to other levels.)
Correlation Study/Agreement with Predicate	No false positives or false negatives compared to predicate.	0 of 20 false positives, 0 of 10 false negatives (compared to competitive device for normal/deficient samples).
Interference	Minimal interference from common substances/conditions.	Bilirubin to 25 mg/dl and lipemia to 1000 mg/dl do not interfere. Hemoglobin is removed by dialysis.

2. Sample Size Used for the Test Set and Data Provenance

Sample Size for Correlation/Comparison Study: 30 samples (16 normal, 4 intermediate, 10 G6PD deficient).
Sample Size for Expected Values:
- Normal population of neonates: 17 observations
- G6PD deficient population: 10 observations
- Intermediate G6PD activity population: 4 observations
Data Provenance: Not explicitly stated (e.g., country of origin). It's reasonable to infer these were collected for the purpose of the study. The study design (clinical samples, comparison to a competitive device) suggests prospective collection or at least retrospective use of stored clinical samples specifically for this performance evaluation.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and the Qualifications of Those Experts

The document does not mention the use of "experts" in the sense of clinicians or radiologists adjudicating results.
The ground truth for the comparison study (normal, intermediate, G6PD deficient) was established by analyzing samples "known to be normal (16), intermediate (4), and G6PD deficient (10)". It implies these classifications were based on prior clinical diagnosis or established laboratory methods, rather than real-time expert consensus for the purpose of this specific study.

4. Adjudication Method for the Test Set

Not applicable/Not mentioned. The study compares the device's results to a "competitive device" and pre-classified samples, rather than an adjudication process among multiple readers for image interpretation or similar tasks.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

No, an MRMC comparative effectiveness study was not done. This device is an in vitro diagnostic reagent kit, not an AI-powered diagnostic system involving human readers.

6. If a Standalone (i.e. algorithm only without human-in-the-loop performance) was done

Yes, the performance characteristics described are for the Astoria-Pacific SPOTCHECK G6PD 50 Hour Reagent Kit as a standalone device, for use by trained laboratory personnel. Its output (µM NADPH) is a quantitative measurement, and its interpretation (e.g., normal, deficient) is based on established ranges. There is no "human-in-the-loop" AI component.

7. The Type of Ground Truth Used

The ground truth for the performance studies was based on:
- Prior clinical classification/diagnosis: Samples were "known to be" normal, intermediate, or G6PD deficient before being tested by the device.
- Comparison to a predicate/competitive device: The device's results were compared against an existing, legally marketed G6PD assay.
- Controlled conditions for precision and linearity: Known concentrations of NADH for linearity, and samples with stable activity levels for precision.

8. The Sample Size for the Training Set

Not applicable. This is not a machine learning or AI device that requires a "training set." It's a chemical reagent kit with established biochemical principles.

9. How the Ground Truth for the Training Set was Established

Not applicable, as there is no training set for this type of device.

Summary

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MAY 1 1 1999

000016

Astoria-Pacific, Inc

G6PD 50-Hour Reagent Kit

510(K) SUMMARY

Name, address, telephone number, contact person and 1. date of preparation of summary.
Applicant's Name and Address Astoria-Pacific, Inc. FDA Establishment No. 3050015 14600 S. E. 82nd Drive Post Office Box 830 Clackamas, OR 97015-0830 USA

1-503-657-3010 TEL 1-503-655-7367 FAX

Raymond. L. Pavitt, President Official Correspondent

Signature of Applicant: Date: March 18, 1999

:
Lesterd Blannion

Garrison, Diagnostics Manager Submission Correspondent

Name of the device, including trade or proprietary 2. name, and classification name.

Product Classification

Product Code Requlation Number 510(k) Number Classification Panel Device Classification 81 JBL 21 CFR 864.7360 K Hematology Kits and Packages Class II

Product Nomenclature

Common Name	G6PD Test
Classification Name	Erythrocytic glucose-6-phosphatedehydrogenase assay, qantitative
Proprietary Name	Astoria-Pacific SPOTCHECKG6PD 50 Hour Reagent Kit
Model Number	Astoria-PacificPart No. 80-3000-13K

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510(K) SUMMARY

Identification of the legally marketed device for which 3. substantial equivalence is claimed.

Product Classification

Product Code	81 JBL
Regulation Number	21 CFR 864.7360
510(k) Number	K790211
S/E Decision Date	March 15, 1979
Classification Panel	Hematology Kits and Packages
Device Classification	Class II

Product Nomenclature

Common Name	G6PD Test
Classification Name	Erythrocytic glucose-6-phosphatedehydrogenase assay, quantitative
Proprietary Name	Glucose Phosphate Dehydrogenase
Model Number	SIGMA Chemical CompanyPreocedure No. 345-UV

4 . Description of the device as found in the labeling

G6PD 50 HOUR REAGENT KIT

API Part No. 80-3000-13K

Glucose-6-Phosphate Dehydrogenase Test System

INTENDED USE

The Astoria-Pacific SPOTCHECK G6PD 50 Hour Reagent Kit is intended for the semi-quantitative determination of glucose-6phosphate dehydrogenase activity in erythrocytes using the Astoria-Pacific SPOTCHECK Analyzer. Measurements of glucose-6-phosphate dehydrogenase activity are used primarily in the diagnosis and treatment of disease states associated with a G6PD deficiency. This method is intended for in vitro diagnostic use primarily as an aid in screening for decreased levels of G6PD enzyme activity. This device is intended for use by trained, qualified laboratory personnel.

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510(K) SUMMARY

Description of the device as found in the labeling (cont.) 4.

SUMMARY AND EXPLANATION OF THE METHOD

G6PD deficiency occurs in many forms. A genetic variant resulting in enzyme instability and mild enzyme deficiency, designated G6PD A, occurs among American blacks at a frequency of about 11%. A mutation that also results in enzyme lability and a much more severe deficiency, designated G6PD Mediterranean, exists in frequencies ranging from <1 to >50% in various Mediterranean populations. Other common variants exist among Asian populations. Such polymorphic enzyme deficiencies are associated with hemolytic anemia during drug administration, infection, and certain other stresses. Less common, functionally more severe mutations result in a chronic hemolytic state even when no abnormal stress is present.

The Astoria-Pacific method is based on the established spectrophotometric methods. Maleimide, an inhibitor of 6phosphoqluconate dehydrogenase (6-PGD) activity, is added to inhibit the production of NADPH by 6-phosphogluconate dehydrogenase.

CHEMICAL PRINCIPLES OF THE PROCEDURE

Enzyme activity is measured by observing NADP+ reduction to NADPH when glucose-6-phosphate is present as a substrate.

G6PD catalyzes the conversion of glucose -6-phosphate (Glu-6-P) to 6-phosphogluconate (6-PG) and, concurrently, the reduction of NADP+ to NADPH.

G6PD Glu-6-P + NADP+ ----------> 6-PG + NADPH + H+ Ma++

The fluorescent NADPH produced is proportional to the G6PD enzyme activity.

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510(K) SUMMARY

Description of the device as found in the labeling (cont.) 4.

Further conversion of 6-PG to ribulose-5'-phosphate (R-5-P) by 6phosphogluconate dehydrogenase (6-PGD) is inhibited by the addition of maleimide.

	6-PGD
6-PG + NADP+	---BLOCKED--->	R-5-P + NADPH + H+ + CO2

REAGENTS

ReagentName	ReactiveIngredient	FinalConcentration
Extraction Buffer	Succinate BufferpH 5.2	6 mM
Tris Buffer	Tris, pH 7.8Triton X-100	50 mM0.5 ml/L
G6PD Substrate	Tris, pH 7.8MaleimideNADPMagnesiumGlu-6-phosphateTRITON X-100	100 mM13 mM1.3 mM1.2 mM1.0 mM0.5 ml/L
NADH Stock Std	TEA Buffer pH 9NADH	50 mM2 mM

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510(K) SUMMARY

Statement of intended use. 5.

Intended Use

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510(K) SUMMARY

A summary of the technological characteristics of the device 6. compared to the predicate device, including chemical composition.

Both devices respond quantitatively to G6PD Activity.

The subject device has the same technological characteristics as the legally marketed predicate device. Specifically, the features, specifications, materials and mode of action are equivalent.

There are no significant differences in technology characteristics between the proposed device and the legally marketed predicate device. The proposed device has the same indications for use as the legally marketed predicate device.

The propose device has the same chemical composition and reaction mechanism as the legally marketed predicate device. There are no new reagents.

The proposed device uses a similar temperature, time and ratio of reagents to sample as the predicate device.

The proposed device uses fluorescence of NADPH to measure G6PD activity in the sample; the predicate device uses UV absorbance of NADPH to measure G6PD activity.

The proposed device is used as an element in a screening strategy that includes other tests and observations and requires confirmation testing and follow-up clinical assesment, as is the predicate device.

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G6PD 50-Hour Reagent Kit

510(K) SUMMARY

A summary of the technological characteristics of the device 6. compared to the predicate device, continued

Comparison Table

Feature	Predicate DeviceSigma Diagnostics	Proposed DeviceAstoria-Pacific Inc
Intended User	Clinical laboratoryprofessionals	Clinical laboratoryprofessionals
Intended Use	Quan titativedetermination of G6PDactivity in bloodred blood cells)	Semi-quan titativedetermination of G6PDactivity in bloodred blood cells)
Indications for use	Screening fordecreased levels ofG6PD activity	Screening fordecreased levels ofG6PD activity
Chemical Principle	NADP+ Reduction toNADPH	NADP+ Reduction toNADPH
Temperature	30° C or 37° C	37° C
Stability	8 hours	8 hours
Expected Value	4.6-13.5 U/g Hb	40-224 µM NADPH
Sensitivity	0.4 U/g Hb	2 µM NADPH
Detection Limit	1.0 U/g Hb	9 µM NADPH
Detection Method	UV absorbanceof NADPH	fluorescence ofNADPH
Wavelength	340 nm	λ excit. =350 nmλ emiss.=450 nm

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Astoria-Pacific, Inc

G6PD 50-Hour Reagent Kit

510(K) SUMMARY

Performance characteristics of the device, including:

Expected values Interfering substances Specific performance characteristics carryover specificity sensitivity within-run precision total precision correlation

EXPECTED VALUES

We have studied a normal population of neonates and these were the values:

17 Number of observations Average Value observed 54 µM Sample Standard Deviation Range of the data >42 µM 95% Confidence interval

150 µM NADPH 40 - 224 μM

We studied a G6PD deficient population, and these were the values:

Number of observations	10
Average Value observed	18 μM NADPH
Sample Standard Deviation	12 μM
Range of the data	4 - 40 μM
95% Confidence interval	0 - 42 μM

We also studied a population with intermediate G6PD activity. These were the values:

Number of observations	4
Average Value observed	49 µM NADPH
Sample Standard Deviation	22 µM
Range of the data	28 - 69 µM
95% Confidence interval	5 - 93 µM

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510(K) SUMMARY

Performance characteristics of the device (continued) 7.
These studies indicate:

Samples exhibiting activity less than about 40 µM NADPH, the lowest observed value of the normal population range, are outside the normal range for G6PD activity and require follow-up and/or additional testing.
There is an overlap in the range for persons with intermediate G6PD activity and the normal population. When that occurs, those samples require follow-up and additional testing.

Each laboratory must determine its own range of normal, intermediate, and deficient levels of G6PD activity, based on its population and analytical variables.

The activity of normal samples varies widely, and the activity of all samples decreases with time under any conditions of storage. Samples showing sufficient activity can be classified as in the normal range. However, the G6PD 50 Hour Reagent Kit can not be used to classify a particular genotype.

Specimens producing abnormal or non-expected responses require confirmation/follow-up testing according to local, state and federal requirements.

Low activity may represent a deteriorated sample. If a sample has deteriorated, or was incorrectly collected, stored or handled, inaccurate results may be obtained.

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510(K) SUMMARY

Performance characteristics of the device (continued) 7.

INTERFERING SUBSTANCES

G6PD is inhibited by NADPH with a K; of 0.02 mM, and by ATP with a Ki of 2 mM 7. Sulfate at 5 mM in vitro causes a decrease of G6PD activity8.

G6PD is increased in cases of pernicious anemia, folic acid deficiency and sickle cell disease; it isn't affected by hairy cell leukemia or multiple sclerosis 9. Copper strongly inhibits this enzyme10.

Neither smoking nor physical training have an effect on the level of enzyme activity10. In one study in the literature, the interindividual variability of G6PD was observed to be 32%, and the intraindividual variation was observed to be 33%.

Hemoglobin may minimally decrease G6PD activity by quenching fluorescence; in this procedure hemoglobin is removed from the reagent stream by dialysis. Bilirubin to 25 mg/dl and lipemia to 1000 mg/dl do not interfere with this test.

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G6PD 50-Hour Reagent Kit

510(K) SUMMARY

Performance characteristics of the device. (continued) 7.

SPECIFIC PERFORMANCE CHARACTERISTICS

Carryover from sample-to-sample is less than 2% and is corrected when a CARRYOVER cup is entered in the sample table.

Specificity for G6PD is achieved by using the specific substrate for the enzyme, glucose-6-phosphate. A small amount of NADPH may be produced by samples in the absence of the substrate.

As little as 2 µM NADPH is discernable from no Sensitivity. response. The response standards from 0 to 75 µm NADH gave a correlation coefficient r > .999 linearity.

Within-run and total precision were evaluated for this Precision. method. Samples with three levels of activity were assayed in duplicate, in 2 runs per day over 5 days to estimate the within-run and total precision. The data is summarized below:

WITHIN-RUN PRECISION (SWR)

	G6PDDeficient	Intermediate	G6PDNormal
Average	7.80 μM	43.0 μM	158 μM
S.D	0.25 μM	1.9 μM	7.34.9 μM
C.V.	3.2 %	4.4 %	3.1 %

TOTAL PRECISION

(ST)

	G6PDDeficient	Intermediate
Average	7.80 μM	43.0 μM
S.D	0.42 μM	2.6 μM
C.V.	5.4 %	6.0 %

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Astoria-Pacific, Inc

G6PD 50-Hour Reagent Kit

510(K) SUMMARY

1. Performance characteristics of the device (continued)
  Correlation. The performance of the G6PD 50-Hour Reagent Kit and a competitive device were evaluated by analyzing samples known to be normal (16), intermediate (4), and G6PD deficient (10). Based upon the evaluation of these 30 samples, a comparison of the performance is presented below:

	FalsePositives	FalseNegatives
API G6PD 50 Hour Kit	0 of 20	0 of 10
Competitive Device	0 of 20	0 of 10

END

510(K) SUMMARY

This 510(k) summary is submitted in accordance with the requirements of 21 CFR § 807.92, as revised April 1, 1998.

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Image /page/12/Picture/1 description: The image shows the logo for the Department of Health & Human Services - USA. The logo is a circular seal with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is an abstract image of an eagle with three lines representing its wings and three wavy lines representing its body.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

MAY 1 1 1999

Mr. Raymond L. Pavitt President Astoria-Pacific, Inc. 14600 S. E. 82nd Drive Post Office Box 830 Clackamas, Origan 97015-0830

K990957 Re:

Trade Name: Astoria-Pacific SPOTCHECK G6PD 50 Hour Reagent Kit Regulatory Class: II Product Code: JBL Dated: March 18, 1999 Received: March 22, 1999

Dear Mr. Pavitt:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic QS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

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Page 2

Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To determine if it does, you should contact the Centers for Disease Control and Prevention (CDC) at (770) 488-7655.

This letter will allow you to begin marketing your device as described in your 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at (301) 594-4588. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at (301) 594-4639. Also, please note the regulation entitled, "Misbranding by reference to premarket notification"(21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll-free number (800) 638-2041 or (301) 443-6597, or at its internet address "http://www.fda.gov/cdrh/dsma/dsmamain.html".

Sincerely yours,

Steven Sutman

Steven I. Gutman, M.D. M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health

Enclosure

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Page / of /

Labels	Values
510(k) Number (if known):	K990957
Device Name:	Spotcheck G6PD
	Kit, so hour.

Indications For Use:

Astoria-Pacific, Inc

G6PD 50-Hour Reagent Kit

INDICATIONS FOR USE

Intended Use

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of Device Evaluation (ODE)

Situe. Maker

(Division Sign-Off)
Division of Clinical Laboratory Devices
510(k) Number K990957

Prescription Use
(Per 21 CFR 801.109)

Over-The-Counter Use_

(Optional Format 1-2-96)

Regulation Number and Section

§ 864.7360 Erythrocytic glucose-6-phosphate dehydrogenase assay.

(a)
Identification. An erythrocytic glucose-6-phosphate dehydrogenase assay is a device used to measure the activity of the enzyme glucose-6-phosphate dehydrogenase or of glucose-6-phosphate dehydrogenase isoenzymes. The results of this assay are used in the diagnosis and treatment of nonspherocytic congenital hemolytic anemia or drug-induced hemolytic anemia associated with a glucose-6-phosphate dehydrogenase deficiency. This generic device includes assays based on fluorescence, electrophoresis, methemoglobin reduction, catalase inhibition, and ultraviolet kinetics.(b)
Classification. Class II (performance standards).