(243 days)
The ESA Biosciences Inc. Vitamin D HPLC test is for the quantitative determination of 25-hydroxyvitamin D in human serum or EDTA-plasma to be used in the assessment of vitamin D sufficiency. Assay results should be used in conjunction with other clinical or laboratory data to assist the clinician in making individual patient management decisions in an adult population.
The ESA method is a complete kit for measurement of Total 25(OH)D by HPLC with electrochemical (EC) detection. Specific reagents and solid phase extraction columns are included for sample preparation and are employed with user-supplied standard laboratory equipment (centrifuge, test tubes, pipettes, etc.). A 200µL volume of sample (serum or plasma) is mixed with a precipitation reagent, which contains internal standard (IS). The internal standard is a stable vitamin D analogue that is used to correct for variability in extraction recovery and analytical sample volume. After centrifugation, supernatant is poured onto a pre-conditioned SPE column for rapid extraction of 25(OH)D and IS. SPE columns are washed with 2 different reagents and analytes are eluted with a third reagent. The resulting eluent is diluted before analysis. The prepared sample is analyzed with an isocratic HPLC system using an ESA EC detector (Coulochem® III or CoulArray®) equipped with a dual coulometric EC cell. Calibration is accomplished by direct HPLC analysis of authentic standard solutions (i.e. not taken through the extraction step). Analysis requires a specific guard and analytical column, mobile phase and calibration reagents to allow rapid quantitative analysis. A dual EC cell is used with the first, upstream, cell maintained at a specific potential to oxidatively screen possible interfering sample components. The second, downstream cell is maintained at a potential that is optimized for selective 25(OH)D detection. The dual coulometric EC cell is a rugged detector that provides much higher selectivity than commonly used absorbance detectors. This allows the use of lower sample volumes than are typically required with HPLC-UV methods and is less susceptible to interferences. Analytical run time is less than 12 minutes and Total 25(OH)D sample concentration is automatically determined by single-point internal standard quantitation.
Here's an analysis of the provided text, outlining the acceptance criteria and study details for the ESA Biosciences Inc. Vitamin D HPLC Test:
1. Table of Acceptance Criteria and Reported Device Performance
The document does not explicitly state "acceptance criteria" in a dedicated section with pass/fail thresholds. However, it presents performance characteristics that implicitly serve as the criteria for evaluating the device's suitability. The table below lists these performance metrics and the reported results.
| Performance Characteristic | Acceptance Criteria (Implied) | Reported Device Performance |
|---|---|---|
| Precision (Within-run) | Low %CV for various samples | Sample #1 (Plasma): 2.82% CV |
| Sample #2 (Plasma): 2.31% CV | ||
| Sample #3 (Serum): 2.55% CV | ||
| Sample #4 (Serum): 2.16% CV | ||
| Level 1: 10.6% CV | ||
| Level 2: 6.8% CV | ||
| Precision (Within-device/Total) | Low %CV for various samples | Sample #1 (Plasma): 6.05% CV |
| Sample #2 (Plasma): 5.48% CV | ||
| Sample #3 (Serum): 5.66% CV | ||
| Sample #4 (Serum): 5.43% CV | ||
| Level 1: 12.6% CV | ||
| Level 2: 8.4% CV | ||
| Limit of Detection (LoD) | Defined as concentration with ≤ 5% false negative/positive | 5.0 ng/mL |
| Limit of Quantitation (LoQ) | % CV ≤ 20% at this concentration | 7.0 ng/mL |
| Limit of Blank (LoB) | N/A (reported as a baseline metric) | 2.5 ng/mL |
| Linearity Range | Linear from LoQ to 200 ng/mL with specific bias goals | Linear from 7.0 ng/mL (LoQ) to 200 ng/mL (bias goals: ≤ 3ng/mL bias for ≤ 20 ng/mL; ≤ 7.5% bias for 21-200 ng/mL) |
| Recovery | Expected % recovery around 100% for spiked samples | Plasma: 90-107% mean recovery |
| Serum: 93-105% mean recovery | ||
| Method Comparison (Correlation) | Correlation Coefficient (R) for comparison with reference method (implies r needs to be within an acceptable range, though r |
§ 862.1825 Vitamin D test system.
(a)
Identification. A vitamin D test system is a device intended for use in clinical laboratories for the quantitative determination of 25-hydroxyvitamin D (25-OH-D) and other hydroxylated metabolites of vitamin D in serum or plasma to be used in the assessment of vitamin D sufficiency.(b)
Classification. Class II (special controls). Vitamin D test systems must comply with the following special controls:(1) Labeling in conformance with 21 CFR 809.10 and
(2) Compliance with existing standards of the National Committee on Clinical Laboratory Standards.