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Standard stock solution: 0.11 mg/mL of USP Minocycline Hydrochloride RS in Medium
Standard solutions: Dilute the Standard stock solution with Medium to obtain solutions with final concentrations of 0.088 mg/mL, 0.0528 mg/mL, 0.0352 mg/mL, 0.022 mg/mL, and 0.0176 mg/mL.
Sample solution: dissolution solution
Injection size: 20 ¦ÌL for the 4 and 24 h time points; 50 ¦ÌL for the 48 and 72 h time points
Analysis: Construct a calibration curve for each sampling interval by plotting the concentration of the
Standard solutions versus peak area. Calculate the slopes and y-intercepts using linear regression analysis.
Calculate the release rate of minocycline:
Resulti = [(rUi − yi)/Si] ¡Á 10/( i ¡Á W ¡Á A)
i =sampling time, 4, 24, 48, 72 h
rUi=peak response from each of the Standard solutions at time i
yi =y-intercept of the calibration curve at sampling time i
Si =slope of the calibration curve at sampling time i
W =weight of the sample (mg)
A =amount of minocycline in the sample (mg/mg of sample) as determined in the Assay
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Tolerances
Time (h)   Release Rate (¦Ìg/h) Average of 6 Measurements
0¨C4       NLT 25
4¨C24      NLT 1.0
24¨C48     NLT 0.2
48¨C72     NLT 0.05

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