Attachment 1. Flowchat of Determination of Vitamin C with HPLC
STANDARD PREPARATION
BEVERAGE SAMPLE
PREPARATION
PREPARATION OF PASTE, POWDER
AND SOLID SAMPLES
Weigh 5 grams of sample (or the
estimated amount of analyte in the
sample) into a 100 ml beaker.
Weigh 10 ml of sample (or
according to the estimated
analyte content in the sample)
into a 100 ml volumetric flask.
Vitamin C standard (ascorbic
acid) 200 mg/l: Weigh 0.01 g of
vitamin C standard (to standard
purity) into a 50 ml volumetric
flask, add 5 ml of 0.1 N HCl,
make up with distilled water.
Stir until dissolved and sonication for
poorly soluble samples
Add 10 ml of 0.1 N HCl
equilibrated with distilled water
up to the mark.
Prepare a working standard
solution of 5, 25, 50, 100, 200
mg/L by pipetting 0.25, 1.25, 2.5,
5, 10 ml of 200 mg/L standard
solution into a 10 ml volumetric
flask, then fill the flask with
distilled water. Place in vial and
inject to HPLC
Using a graduated cup, add 10 mL of 0.1N HCl
and 40 mL of distilled water.
Put into a 100 ml volumetric flask
and add distilled water until the
limit mark, then homogenize.
Put 40 ml of solution into a
centrifuge tube, centrifuge for 5
minutes at 4000 rpm.
Filter the solution through a
0.45 µm, 25 mm RC Syringe
Filter. Transfer the filtered
solution into a vial and inject
to HPLC.
Filter the solution using a 0.45 µm,
25 mm RC syringe filter, then put it
in a vial and inject to HPLC.
This document is for internal use of PT. TÜV NORD Indonesia. Uncontrolled when printed.