SPDS Lutein and Turmeric ERPs

AOAC O FFICIAL M ETHODS OF A NALYSIS (2013)

G UIDELINES FOR D IETARY S UPPLEMENTS AND B OTANICALS Appendix K, p. 15

cancel out. (The Youden ruggedness trial or fractional factorial experiment was designed for this outcome.) Similarly,

Effect of B and b [(x1 + x2 + x5 + x6)/4] – [(x3 + x4 + x7 + x8)/4] = K 4B/4 – 4b/4 = K

These values are plotted on a line. In this case they are more or less uniformly scattered along the line, but some attention should be paid to the extremes. Factor D, the highest positive value represents a difference in solvent, as expected, and this factor has to be investigated further to determine if the high values represents impurities or additional active ingredient. The extreme value of factor G suggests that the extraction should be conducted in the dark. As discussed by Youden, considerably more information can be obtained by utilizing several different materials and several independent replications in different laboratories, so as to obtain an estimate of the standard deviation to be expected between laboratories. Although the ruggedness trial is primarily a method development technique, validation of the application of a method to different matrices and related analytes can be explored simultaneously by this procedure. Comments not used (may be added later): 3.3 Calibration: Run standards from low to high to compensate for any carryover. [Run in random order to compensate for drift is more important than allowing for carryover which should not occur.] Independently made standards results in considerable random error in the calibration curve and is in fact the major source of random error in spectrophotometry. [Therefore a common stock solution is the preferred way of preparing the individual standards.] Version 54 contains revisions as a result of comments from levanseler@nsf.org and McClure. Outline: I. Types and benefits of each method validation study without reproducibility II. Preparing for a Single-Laboratory Method Validation Study III. Review of Performance Characteristics of a Method IV. Errors V. Calibration and Types VI. Bias and Precision Estimations (no reference standard; no reproducibility) VII. Detection and Quantification Limits VIII. Ruggedness

Effect of C and c [(x1 + x3 + x5 + x7)/4] – [(x2 + x4 + x6 + x8)/4] = L 4C/4 – 4c/4 = L

Effect of D and d [(x1 + x2 + x7 + x8)/4] – [(x3 + x4 + x5 + x6)/4] = M 4D/4 – 4d/4 = M

Effect of E and e [(x1 + x3 + x6 + x8)/4] – [(x2 + x4 + x5 + x7)/4] = N 4E/4 – 4e/4 = N

Effect of F and f [(x1 + x4 + x5 + x8)/4] – [(x2 + x3 + x6 + x7)/4] = O 4F/4 – 4f/4 = O

Effect of G and g [(x1 + x4 + x6 + x7)/4] – [(x2 + x3 + x5 + x8)/4] = P 4G/4 – 4g/4 = P

Perform the eight determinations or runs carefully using the assigned factor level combinations and tabulate the values found. Then unscramble the 7 factors and obtain the effect of the assigned factor as the last number. It is important to use the combination of subscripts as assigned for proper interpretation. Expt. Found, % Factors x1 1.03 J (A) = 4A/4 – 4a/4 = 4.86 – 5.14 = –0.28 x2 1.32 K (B) = 4B/4 – 4b/4 = 4.79 – 5.21 = –0.42 x3 1.29 L (C) = 4C/4 – 4c/4 = 4.86 – 5.14 = –0.28 x4 1.22 M (D) = 4D/4 – 4d/4 = 5.05 – 4.95 = +0.10 x5 1.27 N (E) = 4E/4 – 4e/4 = 4.92 – 5.08 = –0.16 x6 1.17 O (F) = 4F/4 – 4f/4 = 4.95 – 5.05 = –0.10 x7 1.27 P (G) = 4G/4 – 4g/4 = 4.69 – 5.31 = –0.62 x8 1.43

© 2013 AOAC INTERNATIONAL