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Example: AOAC Official Method 2015.11
Chondroitin Sulfate Content in Raw Materials and Dietary Supplements
High‐Performance Liquid Chromatography with Ultraviolet Detection After Enzymatic Hydrolysis
First Action 2015
A. Method Performance
The performance criteria in Table 2015.11A is prescribed by
Standard Methods Performance Requirements
(SMPR)
2014.009
, which was adopted by the AOAC Stakeholder Panel for Dietary Supplements (SPDS). Laboratory(ies)
implementing this method should follow ISO Standard 17025 to verify that the performance of this method in your
laboratory meets the criteria in table 2015.11A. Please report verification results to AOAC at this link:
AOAC
2015.11
.
Table 2015.11A: Verification Acceptance Criteria
Analytical Ranges
Parameters
1‐10% (w/w)
>10‐100% (w/w)
Repeatability (RSD
r
)
≤ 3%
≤2%
Recovery
92‐105%
98‐102%
B. Materials
Chondroitin sulfate (CS) raw materials from bovine trachea, porcine skin/cartilage, and shark cartilage, and CS
control material from bovine trachea were obtained from Bioiberica (Barcelona, Spain). Dietary supplement
products containing CS (hard‐shell capsules, tablets, chewables, softgels, and liquids) were obtained from
commercial suppliers. Descriptions of the dietary supplement products used in the study are presented in Table
2015.11B
.
C. Apparatus
(
a
)
LC system
.—Beckman 126 dual high pressure mixing pumps (Beckman Coulter, Fullerton, CA, USA), 168 diode
array UV detector, 507e autosampler, and 32 Karat software.
(
b
)
Operating conditions
.—Mobile phase flow rate, 1.1 mL/min; column temperature, ambient; injection volume,
30 μL; and detection, 240 nm.
(
c
)
LC column
.—Phenomenex Synergi Polar‐RP, 4.6 × 150 mm, 4 μm particle size (Phenomenex, Torrance, CA, USA).
(
d
)
Analytical balance
.—Accu‐124 (Fisher Scientific, Pittsburgh, PA, USA), ±0.01 mg readability.
(
e
)
Ultrasonic bath
.—Model FS60H (Fisher Scientific).
(
f
)
pH meter
.—Model pH 500 (Oakton, Vernon Hills, IL, USA), ±0.01 pH unit readability.
(
g
)
Dry block heater
.—Isotemp Dry Bath Incubator (Fisher Scientific), maintained at 37°C.
(
h
)
LC injection vials
.—2 mL, with caps and Teflon‐coated septa.
(
i
)
Limited volume inserts
.—200 μL, for LC vials.
(
j
)
Syringes
.—25, 100, and 500 μL Luer‐Lok.
D. Reagents
Note
: Chemicals from other suppliers meeting the specificationsmay also be used.
(
a
)
Solvents
.—Acetonitrile, LC grade; water, LC grade; hydrochloric acid, concentrated, ACS reagent grade.
AOAC OMB Meeting
February 10-11, 2016
48