.6. Method validation
A recovery study for each vitamin was performed as follows
using Spearmint as model sample: 2 sets of extractions were carried
out simultaneously following the optimized procedure. In one
of them, the sample spiked with a known amount of the target compounds
was extracted whereas in the other, the same sample was
extracted and spiked with the same concentration of vitamins after
extraction. Peak areas of vitamins from the same sample spiked
before and after the extraction process were compared. The recovery
study was performed in three different days, always with the
same sample in order to check its repeatability.
Once optimum separation conditions and MS/MS detection
parameters were optimized, instrumental intraday and interday
precision was assessed. For this purpose, the mixture of the studied
vitamins was consecutively injected five times in the same day
(n = 5) and also in three different days (n = 15). The vitamins concentration
of this mixture was in the middle range of the linearity
of each one (vitamin C: 1.32 g/ml; B1: 0.24 g/ml; B2: 0.3 g/ml;
B3: 0.7 g/ml; B5: 0.33 g/ml; B6: 0.27 g/ml; B9: 0.13 g/ml). Limits
of detection (LOD) and quantification (LOQ) were calculated as
a signal-to-noise ratio of 3 and 10, respectively, and expressed in
ng/ml. Accuracy of the method was assessed by using the recovery
percentages of spiked samples.
.6. Method validation
A recovery study for each vitamin was performed as follows
using Spearmint as model sample: 2 sets of extractions were carried
out simultaneously following the optimized procedure. In one
of them, the sample spiked with a known amount of the target compounds
was extracted whereas in the other, the same sample was
extracted and spiked with the same concentration of vitamins after
extraction. Peak areas of vitamins from the same sample spiked
before and after the extraction process were compared. The recovery
study was performed in three different days, always with the
same sample in order to check its repeatability.
Once optimum separation conditions and MS/MS detection
parameters were optimized, instrumental intraday and interday
precision was assessed. For this purpose, the mixture of the studied
vitamins was consecutively injected five times in the same day
(n = 5) and also in three different days (n = 15). The vitamins concentration
of this mixture was in the middle range of the linearity
of each one (vitamin C: 1.32 g/ml; B1: 0.24 g/ml; B2: 0.3 g/ml;
B3: 0.7 g/ml; B5: 0.33 g/ml; B6: 0.27 g/ml; B9: 0.13 g/ml). Limits
of detection (LOD) and quantification (LOQ) were calculated as
a signal-to-noise ratio of 3 and 10, respectively, and expressed in
ng/ml. Accuracy of the method was assessed by using the recovery
percentages of spiked samples.
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