This European Standard specifies a method for the determination of vitamin B1 in food by high performance liquid chromatography (HPLC) with enzymatic treatment and pre- or post-column derivatization. This method has been validated in two interlaboratory studies. The first study was for the analysis of samples of whole meal flour, milk powder\/spray dried milk, freeze-dried mixed vegetables and freeze-dried pig’s liver ranging from 0,295 mg\/100 g to 0,807 mg\/100 g. The second study was for the analysis of samples of tube feeding solution, baby food with vegetables, powdered milk, meal with fruits, yeast, cereal, chocolate powder and food supplement ranging from 0,11 mg\/100 g to 486 mg\/100 g. Vitamin B1 is the mass fraction of total thiamin including its phosphorylated derivatives.<\/p>\n
For further information on the validation, see Clause 8 and Annex B.<\/p>\n
| PDF Pages<\/th>\n | PDF Title<\/th>\n<\/tr>\n | ||||||
|---|---|---|---|---|---|---|---|
| 5<\/td>\n | Foreword <\/td>\n<\/tr>\n | ||||||
| 6<\/td>\n | 1 Scope 2 Normative references 3 Principle 4 Reagents <\/td>\n<\/tr>\n | ||||||
| 9<\/td>\n | 5 Apparatus <\/td>\n<\/tr>\n | ||||||
| 10<\/td>\n | 6 Procedure 6.1 Preparation of the test sample 6.2 Preparation of the sample test solution 6.2.1 Extraction 6.2.2 Enzyme treatment 6.2.3 Sample test solution 6.3 Oxidation of thiamin to thiochrome 6.3.1 General <\/td>\n<\/tr>\n | ||||||
| 11<\/td>\n | 6.3.2 Pre-column oxidation 6.3.2.1 Procedure for oxidation step 6.3.2.2 Identification with HPLC after pre-column oxidation 6.3.3 Post-column oxidation 6.3.3.1 Procedure for oxidation step 6.3.3.2 Identification with HPLC using post-column oxidation <\/td>\n<\/tr>\n | ||||||
| 12<\/td>\n | 6.4 Determination 7 Calculation <\/td>\n<\/tr>\n | ||||||
| 13<\/td>\n | 8 Precision 8.1 General 8.2 Repeatability 8.3 Reproducibility <\/td>\n<\/tr>\n | ||||||
| 14<\/td>\n | 9 Test report <\/td>\n<\/tr>\n | ||||||
| 15<\/td>\n | Annex A (informative) Examples of HPLC chromatograms Figure A.1 \u2014 Example for an HPLC separation of thiamin as thiochrome standard using pre-column derivatization <\/td>\n<\/tr>\n | ||||||
| 16<\/td>\n | Figure A.2 \u2014 Examples for a HPLC separation of thiamin using post-column derivatization in lettuce (a), cooked rice (b) and cooked pork meat (c) <\/td>\n<\/tr>\n | ||||||
| 17<\/td>\n | Annex B (informative) Precision data Table B.1 \u2014 Precision data for whole meal flour, milk powder\/spray dried milk, freeze-dried mixed vegetables and freeze-dried pig’s liver <\/td>\n<\/tr>\n | ||||||
| 18<\/td>\n | Table B.2 \u2014 Precision data for tube feeding solution, baby food with vegetables, powdered milk, meal with fruits and yeast <\/td>\n<\/tr>\n | ||||||
| 19<\/td>\n | Table B.3 \u2014 Precision data for cereal, chocolate powder and food supplement <\/td>\n<\/tr>\n | ||||||
| 20<\/td>\n | Annex C (informative) Alternative HPLC systems Table C.1 \u2014 Alternative HPLC conditions <\/td>\n<\/tr>\n | ||||||
| 21<\/td>\n | Annex D (informative) Vitamin B1 compound: 2-(1-hydroxyethyl)thiamin (HET) performing post-column derivatization <\/td>\n<\/tr>\n | ||||||
| 22<\/td>\n | Figure D.1 \u2014 Chromatogram of an extract of CRM 487 pigs liver (a) and a standard of 0,1 \u00b5g\/ml TMP (Thiamin Mono Phosphate), 0,1 \u00b5g\/ml thiamin and 0,01 \u00b5g\/ml HET (2-(1-hydroxyethyl)thiamin) (b) <\/td>\n<\/tr>\n | ||||||
| 23<\/td>\n | Bibliography <\/td>\n<\/tr>\n<\/table>\n","protected":false},"excerpt":{"rendered":" Foodstuffs. Determination of vitamin B1 by high performance liquid chromatography<\/b><\/p>\n |