Chemical structures and descriptions

Vitamin B2

B2

What is measured?

Synonym: Riboflavin.
What is measured: Riboflavin and the flavin cofactor, flavin mononucleotide (FMN).
Method(s): LC-MS/MS (1).

Indication(s)

Riboflavin is a marker of vitamin B2 status (2).

Specimen, collection and processing

Patient/subject: Prandial status may have some effect on concentrations of riboflavin, but has no effect on FMN.
Matrix: Riboflavin and FMN are most stable in EDTA plasma.
Volume: Minimum volume is 60 µL, but 200 µL is optimal and allows reanalysis.
Preparation and stability: The plasma fraction must be separated from the blood cells, preferentially within 30 minutes. Samples should be put on ice, and protected from light. Riboflavin is light sensitive. In serum, citrate plasma or heparin plasma, FMN is converted to riboflavin. Measurement of the sum of riboflavin plus FMN is an option in samples that have not been adequately treated.

Transportation

Frozen, on dry ice. (for general instruction on transportation, click here.)

Reported values, interpretation

Reported values (riboflavin/FMN): 5-100/3-30 nmol/L.
Intraclass correlation coefficient (ICC) (riboflavin/FMN): 0.79 /0.69.

Literature

1. Midttun, O., Hustad, S., and Ueland, P.M. (2009). Quantitative profiling of biomarkers related to B-vitamin status, tryptophan metabolism and inflammation in human plasma by liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Sp 23, 1371-79.
2. Hustad, S., McKinley, M.C., McNulty, H., Schneede, J., Strain, J.J., Scott, J.M., and Ueland, P.M. (2002). Riboflavin, flavin mononucleotide, and flavin adenine dinucleotide in human plasma and erythrocytes at baseline and after low-dose riboflavin supplementation. Clin Chem 48, 1571-77.

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Statistical power is the probability that a statistical test will correctly reject a false null hypothesis (H0​) when a specific alternative hypothesis (H1​) is true. H0​ is the null hypothesis, which states there is no effect or no difference. H1​ is the alternative hypothesis, which states there is a real effect or difference. Alpha (α) is the probability of a Type I error (a false positive), which is the risk of incorrectly rejecting the H0​ when it is actually true. You set this value before the experiment, commonly at 0.05. Beta (β) is the probability of a Type II error (a false negative), which is the risk of failing to reject the H0​ when it is actually false.

Power is calculated as 1−β. Increasing power means you are decreasing the probability of making a Type II error.

Several factors can be adjusted to increase the power of a statistical test:

  • Effect Size: This is the magnitude of the difference you are trying to detect. A larger effect size is easier to detect, thus increasing power. 

  • Sample Size: The number of observations in a study. A larger sample size provides more information about the population, reducing the margin of error and increasing the power to detect a true effect.

  • Variation: Refers to the spread or standard deviation of the data within the population. Less variation makes it easier to distinguish a real effect from random noise, thereby increasing power.

  • Alpha (): Increasing the alpha level (e.g., from 0.05 to 0.10) also increases power, but at the cost of a higher risk of a Type I error. This trade-off is often undesirable.

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