SimultaneousDetermination of SevenWater SolubleVitamins inProducts byLC-MS/MS
Page
16
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20
TableA2. WorkingStandard solutionpreparation.
Solution
IWSS
Volume
(µL)
Recommended
MaximumPipette
Volume
ISSM
Volume
(µL)
Recommended
Maximum
PipetteVolume
Final
Volume
(mL)
WS1
10
10
50
100
25
WS2
50
100
50
100
25
WS3
200
250
50
100
25
WS4
500
1000
50
100
25
WS5
1000
1000
50
100
25
ii)
Adjust pHwith additionof 80µL of 30% ammoniumhydroxide, cap, andmix
n)
Procedure
i)
Powder Products
(1)
Powder products are reconstitutedwithwater prior to analysis
ii)
SampleAnalysis
(1)
Using a tarred, 50-mL, self-standing centrifuge tube, weigh the appropriate sample amount.
(2)
Add50µLof the internal standardmixture (ISSM) via positive-displacement pipette.
(3)
Vortex the sample briefly (approx. 10 seconds) tomix in the ISSM.
(4)
Add25±2mLof 1% ascorbic acid to the sample. Abottle topdispenser can beused.
(5)
Tightly cap the tube andmix the sample by inverting aminimumof 5 times.
(6)
Add80µL30% ammoniumhydroxide to the sample via positive displacement pipette.
(7)
Tightly cap the sample tube andmix thoroughlyby inverting the tube aminimumof 5 times.
(8)
Allow the sample to stand for aminimumof 10minutes but not longer than60minutes.
(9)
Filter an aliquot of the sample extract into an amber autosampler vial using a 0.45µmPTFE
syringe filter.
(10)
For theAnalysis of pantothenic acid, niacin, pyridoxine, thiamine, and riboflavin (1-16
dilution)
(a)
Pipette 1500µLof theDiluent Solution into a clean autosampler vial
(b)
Pipette 100µLof the filtered sample (step6.i) into the vial with the diluent, cap and invert 5
to7 times tomix.
(i)
Expiration:
Samples prepared in 1% ascorbic acid solution have been determined to
be stable for up to72 hours at room temperature. Working standard solutions and
samplesmust be tested as a complete set if reanalysis is required.
(11)
For theAnalysis of folic acid andbiotin (1-4dilution)
(a)
Pipette 750µLof theDiluent Solution into a clean autosampler vial
(b)
Pipette 250µLof the filtered sample (step6.i) into the vial with the diluent, cap and invert 5
to 7 times tomix.
(i)
Expiration:
Samples prepared in 1% ascorbic acid solutionhave been determined to
be stable for up to72hours at room temperature. Working standard solutions and
samplesmust be tested as a complete set if reanalysis is required.
BIO-03
FORWORKINGGROUP/ERPUSEONLY
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