HICHEM ALP/AMP REAGENT KIT by HICHEM DIAGNOSTICS

HiChem ALP/AMP Reagent (product no. 70001) is intended for the quantitative determination of alkaline phosphatase in serum and plasma. The principal diagnostic indications of elevated serum alkaline phosphatase are diseases of the liver, bone, parathyroid and intestine.

Device Description

The HiChem ALP/AMP Reagent determines alkaline phosphatase by enzymatic hydrolysis of p-nitrophenylphosphate to p-nitrophenoxide at alkaline pH. The HiChem ALP/AMP Reagent is intended to be used either as a manual procedure or on clinical analyzers which can automate the required manipulations. The reagent is supplied as two liquid-stable components which are combined, either before or during use, in the approximate ratio of 1 part ALP/AMP Substrate and 5 parts ALP/AMP Reagent Buffer. The ALP/AMP Substrate can also be used as a start reagent and combined with the Reagent Buffer following sample addition.

AI/ML Overview

Here's a breakdown of the acceptance criteria and study information for the HiChem ALP/AMP Reagent, based on the provided text:

Important Note: The provided document is a 510(k) summary for a diagnostic reagent, not a medical device in the typical sense of AI-powered imaging or diagnostic software. Therefore, many of the requested categories (e.g., number of experts, adjudication methods, MRMC studies, standalone performance for AI, training set details) are not applicable to this type of submission. This document focuses on demonstrating substantial equivalence to existing, legally marketed devices through analytical performance characteristics.

Acceptance Criteria and Reported Device Performance

Device: HiChem ALP/AMP Reagent (product no. 70001)

Intended Use: Quantitative determination of alkaline phosphatase (ALP) in serum and plasma.

Acceptance Criteria Category	Acceptance Criteria (Stated or Implied)	Reported Device Performance
Manual Procedure
Linearity (30°C)	Linear to at least 900 U/L	Linear to at least 900 U/L (Recoveries at 30°C) = -1.7 U/L + 1.005 x (Standard Activity), r² = 0.9998, sy.x = 6.5 U/L
Linearity (37°C)	Linear to at least 900 U/L	Linear to at least 900 U/L (Recoveries at 37°C) = 2.9 U/L + 0.969 x (Standard Activity), r² = 0.9998, sy.x = 8.0 U/L
Precision (37°C)	Replicate assay with acceptable within-run and total SD values (implied by tabulated results)	See table below for detailed precision results at 37°C (n=30 for each control).
Comparison to MAS ALP Reagent	Acceptable correlation and agreement (implied by regression statistics)	r² = 0.9986, (HiChem Results) = -0.2 U/L + 1.058 x (MAS Results), sy.x = 5.5 U/L
Anticoagulant Interference	Bias < 2% and statistically insignificant at 95% confidence level for heparin and lithium iodoacetate	Bias < 2% and statistically insignificant at 95% confidence level for heparin and lithium iodoacetate
Combined Reagent Stability (2-8°C)	Shifts in recovery < 3 U/L or 5% (whichever is greater) over 1 month	Shifts in recovery < 3 U/L or 5% (whichever is greater) over 1 month
Combined Reagent Stability (18-25°C)	Shifts in recovery < 3 U/L or 5% (whichever is greater) over 3 days	Shifts in recovery < 3 U/L or 5% (whichever is greater) over 3 days
Automated Procedure (Hitachi 704)
Linearity	Linear to at least 1,200 U/L	Linear to at least 1,200 U/L (Recoveries) = 3.6 U/L + 0.981 x (Standard Activity), r² = 1.000, sy.x = 3.6 U/L
Precision	Replicate assay with acceptable within-run and total SD values (implied by tabulated results)	See table below for detailed precision results (n=60 for each control).
Comparison to BMD ALP/AMP Reagent	Acceptable correlation and agreement (implied by regression statistics)	r² = 1.000, (HiChem Results) = 1.2 U/L + 1.974 x (BMD Results), sy.x = 1.4 U/L
Calibration Stability (24 hours)	Observed shifts in recoveries < 0.25%	Observed shifts in recoveries < 0.25% over 24 hours without calibration
On-board Stability (2 weeks)	Largest observed control shift < 2% over 15 days	Largest observed control shift was 2% over 15 days

Precision Results - Manual Procedure (37°C):

Specimen	n	mean	within-run SD	total SD
Serum control 1	30	55 U/L	2.1 U/L	2.2 U/L
Serum control 2	30	225 U/L	3.7 U/L	3.9 U/L
Serum control 3	30	765 U/L	6.6 U/L	9.0 U/L

Precision Results - Automated Procedure:

Specimen	n	mean	within-run SD	total SD
Serum control 1	60	49 U/L	0.4 U/L	0.6 U/L
Serum control 2	60	196 U/L	0.8 U/L	1.1 U/L
Serum control 3	60	692 U/L	2.5 U/L	3.0 U/L

Study Details

Sample size used for the test set and the data provenance:
- Manual Procedure:
  - Linearity: Standards ranging from 0 to over 1,350 U/L (number of individual standards not specified, but usually involves multiple levels).
  - Precision: 30 replicates for each of 3 control sera.
  - Method Comparison (vs. MAS Reagent): 83 mixed serum and plasma specimens.
  - Anticoagulant Interference: "spiked serum pools" (specific number not given, but plural implies more than one).
  - Stability: Serum controls (ranging from approx. 50 to 750 U/L ALP) tested over 1 month and 3 days.
- Automated Procedure (Hitachi 704):
  - Linearity: Ten linearity standards spanning the claimed linear range.
  - Precision: 60 replicates for each of 3 commercially available control sera.
  - Method Comparison (vs. BMD Reagent): 178 mixed serum and plasma specimens.
  - Calibration Stability: Serum controls (ranging from approx. 50 to 750 U/L ALP).
  - On-board Stability: Serum controls using the same reagent tested over 15 days.
- Data Provenance: Not explicitly stated (e.g., country of origin). The studies appear to be prospective analytical performance studies designed to test the reagent's characteristics. The "mixed serum and plasma specimens" for method comparison suggest real-world patient samples.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- N/A. This is an in-vitro diagnostic reagent. Ground truth is established through the measurement of analyte concentrations using the stated methods and comparisons to established, legally marketed predicate devices, not through expert human interpretation.
Adjudication method (e.g., 2+1, 3+1, none) for the test set:
- N/A. Adjudication methods are relevant for subjective interpretations (e.g., medical imaging reads), not for quantitative chemical measurements.
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:
- N/A. This applies to AI-assisted diagnostic tools, not chemical reagents.
If a standalone (i.e. algorithm only without human-in-the loop performance) was done:
- N/A. This applies to AI algorithms, not chemical reagents. The performance described is the "standalone" performance of the reagent, either manually or on an automated analyzer.
The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
- Analytical Measurement (Reference Methods/Predicate Devices): The "ground truth" for the linearity studies were known concentrations of alkaline phosphatase in standards. For method comparison studies, the "ground truth" was established by measurements performed using the legally marketed predicate devices (MAS ALP Reagent and BMD Alkaline Phosphatase/AMP Reagent), which are accepted reference points for determining substantial equivalence.
The sample size for the training set:
- N/A. This reagent is not an AI algorithm requiring a training set in the conventional sense. The "training" of the reagent would involve its formulation and optimization during development, which is not detailed here.
How the ground truth for the training set was established:
- N/A. See point 7.

Summary

{0}------------------------------------------------

/963383 SEP

DIAGNOSTICS

SUMMARY OF 510(K) SAFETY AND EFFECTIVENESS INFORMATION

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The HiChem ALP/AMP Reagent is substantially equivalent to the BMD Alkaline Phosphatase/AMP Reagent, product no. 704093, manufactured by Boehringer Mannheim Corp., Indianapolis, IN., and the MAS ALP Reagent, product no. 139-154 manufactured by Medical Analysis Systems, Inc., Camarillo, CA. All three reagents support the same intended use and produce equivalent results with the same clinical purpose. In addition, they are all based on the same methodology which determines alkaline phosphatase (ALP) through the measurement of liberated p-nitrophenoxide. Finally, all reagents are sold in a generic format which supplies the reagent with a manual procedure and supports its use on various instruments through procedure supplication sheets).

The effectiveness of the manual procedure is shown by the recovery of linearity standards, the precision of control recoveries, the comparison of serum and plasma recoveries to the MAS ALP Reagent and the validation of the chemical additives and reconstituted stability claims.

The recovery of alkaline phosphatase using HiChem ALP/AMP Reagent as a manual method at both 30°C and 37 C reaction temperatures is linear to at least 900 U/L as shown by the recovery of linearity standards ranging from 0 to over 1,350 U/L. Regression statistics are shown below.

(Recoveries at 30°C) = -1.7 U/L + 1.005 x (Standard Activity),	$r^2$ = 0.9998,	sy.x = 6.5 U/L
(Recoveries at 37°C) = 2.9 U/L + 0.969 x (Standard Activity),	$r^2$ = 0.9998,	sy.x = 8.0 U/L

Precision, demonstrated by replicate assay of control sera at 37°C, is shown below.

Specimen	n	mean	within-run SD	total SD
Serum control 1	30	55 U/L	2.1 U/L	2.2 U/L
Serum control 2	30	225 U/L	3.7 U/L	3.9 U/L
Serum control 3	30	765 U/L	6.6 U/L	9.0 U/L

Alkaline phosphatase recoveries of 83 mixed serum and plasma specimens are compared between the HiChem and MAS reagents. Least squares regression statistics are shown below.

$r^2$ = 0.9986, (HiChem Results) = -0.2 U/L + 1.058 x (MAS Results) sv.x = 5.5 U/L.

The use of heparin and lithium iodoacetate as anticoagulants is shown by the assay of spiked serum pools. In all cases, the bias due to the addition of anticoagulant is less than 2% and statistically insignificant at a 95% confidence level.

The stability of the combined working reagent over 1 month at 2-8 C and 3 days at 18-25 C are documented through the recovery of serum controls which range from approximately 50 to 750 U/L AMP at 37°C. For all controls, the observed shifts in recovery were less than the greater of 3 U/L or 5%.

The effectiveness of the automated Hitachi 704 procedure is shown by the recovery of linearity standards, the precision of control recovery of serum controls over both the calibration stability period and the onboard stability claim, and the comparison of patient specimen recoveries to the BMD Alkaline Phosphatase/AMP Reagent.

510(k) Notification, HiChem ALP/AMP Reagent Kit HiChem Diagnostics, Brea, California

23 August, 1996 Page 53 of 54

{1}------------------------------------------------

The recovery of alkaline phosphatase using HiChem ALP/AMP Reagent as an automated method is linear to at least 1.200 UL as shown by the recovery of ten linearity standards which span the claimed linear range. Regression statistics are shown below.

$$\text{//}\text{\'e\`e`em,\text{"Rec`ever`ies`es}\text{`es}}\text{`a``ed'`e``ed'`e} = \text{3.6```ed'`L} + \text{0.98``I} \times \text{\text{`a`et`b`e`d`e`e}}\text{`a``ed'`e}, \qquad \text{#\text{%} = \text{1.000}, \qquad \text{#\text{%} = \text{3.6``ed'de`e}}\text{\text{#\text{%} = \text{1.000}}}$$

Precision, demonstrated by replicate assay of commercially available control sera, is shown below.

Specimen	n	mean	within-run SD	total SD
Serum control 1	60	49 U/L	0.4 U/L	0.6 U/L
Serum control 2	60	196 U/L	0.8 U/L	1.1 U/L
Serum control 3	60	692 U/L	2.5 U/L	3.0 U/L

Alkaline phosphatase recoveries of 178 mixed serum and plasma specimens compared between the HiChem and BMD reagents using least squares regression, vield the following statistics.

$r^2$ = 1.000, (HiChem Results) = 1.2 U/L + 1.974 x (BMD Results) sv.x = 1.4 U/L.

The 24 hour calibration stability claim is documented through the recovery of serum controls which span from approximately 50 to 750 U/L ALP. In all cases, the observed shifts in recoveries over 24 hours without calibration are less than 0.25%. The on-board stability claim of 2 weeks is documented by the recovery of serum controls using the same reagent, left on the analyzer, over 15 days. The largest observed control shift over the 15 day period was only 2%.

The HiChem ALP/AMP Reagent is shown to be safe and effective and substantially equivalent to the BMD Alkaline Phosphatase/AMP Reagent, product no. 704093, manufactured by Boehringer Mannheim Corp., Indianapolis, IN., and the MAS ALP Reagent, product no. 139-154 manufactured by Medical Analysis Systems, Inc., Camarillo, CA.

Wynn Stocking

Wynh Stocking Manager of Regulatory Affairs HiChem Diagnostics

Regulation Number and Section

§ 862.1050 Alkaline phosphatase or isoenzymes test system.

(a)
Identification. An alkaline phosphatase or isoenzymes test system is a device intended to measure alkaline phosphatase or its isoenzymes (a group of enzymes with similar biological activity) in serum or plasma. Measurements of alkaline phosphatase or its isoenzymes are used in the diagnosis and treatment of liver, bone, parathyroid, and intestinal diseases.(b)
Classification. Class II.