SimultaneousDetermination of SevenWater SolubleVitamins inProducts byLC-MS/MS
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Call forMethods:Biotin
MethodTitle:
SimultaneousDeterminationof SevenWater SolubleVitamins inProducts byLC-MS/MS
MethodAuthors:
JosephThompson, Ph.D.,Marti Cenky, JamesDenison, C. Thomas Seipelt, AdamSchuh,
MelissaNixon, NicholasCellar, Ph.D., DonGilliland, Ph.D., JasonHolton, Abbott Nutrition, Columbus, OH
USA
Compounds determined
: Biotin, FolicAcid, PantothenicAcid, Niacin, Pyridoxine, Thiamine, Riboflavin
Methoddescription:
Samples are prepared by the addition of 1% ascorbic acid topartiallyprecipitate proteins
aswell as protect folic acid fromdegradation. Stable-isotope labeled internal standards are incorporated into the
sample preparation and are used to correct for variability inboth the sample preparation and instrument operation.
Prepared samples andworking standard solutions are injectedontoultra performance liquid chromatograph
(UPLC) interfaced to a triple-quadrupolemass spectrometer (MS/MS) for analysis.
Status of thismethod:
method is validated and in-use at Abbott central R&D facility inColumbus
applicabilityof thismethod includes the six listedwater soluble vitamins in addition to biotin listed in the
call formethods.
LimitedSLV performedonSPIFANmatrices (data presented below)
SMPRRequirements
(fromSPIFANwebsite)
Biotin, folic acid (as total folate) andpantothenic acidhave definedSMPR requirements. As SMPR’s have not
yet been established for niacin, pyridoxine, riboflavin and thiamine themethod performance for those compounds
will be compared to theSMPR for biotin.
*
μg /100g reconstituted final product.
Table 1.Method performance requirements for the analysis of biotin
Analytical range
0.1 - 150
*
Limit ofQuantitation (LOQ)
≤ 0.1
*
Repeatability (RSDr)
0.1-1
*
≤ 8%
>1
*
≤ 6%
Recovery
0.1-1
*
80% to120%ofmean
spiked recoveryover the range of
the assay.
>1
*
90% to 110%ofmean spiked
recoveryover the range of the
assay.
Reproducibility (RSDR)
0.1-1
*
≤ 16%
>1
*
≤ 12%
Concentrations apply to: a) ‘ready-to-feed” liquids “as is”; b) re-
constituted powders (25 g into 200 gofwater); and c) liquid
concentrates diluted 1:1 byweight.
BIO-03
FORWORKINGGROUP/ERPUSEONLY
DONOTDISTRIBUTE
