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GB/T 5009.197-2003 (GB/T5009.197-2003)

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GB/T 5009.197-2003: PDF in English (GBT 5009.197-2003)

GB/T 5009.197-2003
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
ICS 67.040
C 53
Determination of thiamine hydrochloride, pyridoxine
hydrochloride, niacin, niacinamide and caffeine in
health foods
ISSUED ON. AUGUST 11, 2003
IMPLEMENTED ON. JANUARY 01, 2004
Issued by. Ministry of Health of the People's Republic of China;
Standardization Administration of the People's Republic of
China.
Table of Contents
Foreword ... 3 
Introduction ... 4 
1  Scope ... 5 
2  Principle ... 5 
3  Reagents ... 5 
4  Instruments ... 6 
5  Analysis procedures ... 6 
6  Result calculation ... 8 
7  Precision ... 8 
Introduction 
Thiamine hydrochloride, pyridoxine hydrochloride, niacin, niacinamide and other
vitamins-B family are the indispensable nutrients for human beings; they play an important
role in prevention and treatment of various diseases. Currently vitamins-B family and
caffeine are added into health food as functional component. On basis of the reference to
the vitamin detection method of Japanese Taisho pharmaceutical Co., Ltd, and combined
with the specific conditions of instrument analysis in our country, this method is formulated
as a high performance liquid chromatographic method that can determine the thiamine
hydrochloride, pyridoxine hydrochloride, niacin, niacinamide and other vitamins-B family
and caffeine that are added in tablet, capsule, oral liquid, beverages as well as other
health foods.
Determination of thiamine hydrochloride, pyridoxine
hydrochloride, niacin, niacinamide and caffeine in
health foods
1  Scope 
This Standard specifies the high performance liquid chromatography determination
method of thiamine hydrochloride, pyridoxine hydrochloride, niacin, niacinamide and
caffeine in health foods.
This Standard is applicable to the high performance liquid chromatography determination
method for ADDING thiamine hydrochloride, pyridoxine hydrochloride, niacin, niacinamide
and caffeine that are served as dietary supplements INTO test sample types such as
tablet, capsule, oral liquid and drink.
Detection limit of this method is. 0.02μg for thiamine hydrochloride, 0.02μg for pyridoxine
hydrochloride, 0.10μg for niacin, 0.06μg for niacinamide, and 0.05μg for caffeine. The
linear range is. 0.500μg/mL ~ 50.0μg/mL for thiamine hydrochloride; 0.500μg/mL ~
50.0μg/mL for pyridoxine hydrochloride; 1.00μg/mL ~ 100μg/mL for niacin; 1.00μg/mL ~
100μg/mL for niacinamide; and 5.00μg/mL ~ 500μg/mL for caffeine.
2  Principle 
Use methanol + water + phosphoric acid=100+400+0.5 to extract and dilute the uniformly
crushed tablet, capsule or liquid test samples; then conduct qualitative and quantitative
determination according to the external standard method of high performance liquid
chromatography UV detector.
3  Reagents 
Unless otherwise specified, the water used in the analysis is double-distilled water.
3.1 Methanol. Guaranteed reagent.
3.2 Acetonitrile. Chromatographically pure.
3.3 Phosphoric acid. Analytically pure.
3.4 Sodium dodecylsulhte. High performance liquid chromatography (HPLC) reagents.
3.5 Sodium 1-decanesulfonate. High performance liquid chromatography (HPLC)
reagents.
3.6 Standard stock solution A. Accurately and respectively weigh 0.0500g of standard
thiamine hydrochloride and 0.0500g of standard pyridoxine hydrochloride of which the
moisture has been deducted; dissolve into water and fix the volume to 50mL.
3.7 Standard stock solution B. Accurately and respectively weigh 0.0400g of standard
nicotinamide and 0.0400g of standard niacin of which the moisture has been deducted;
dissolve into water and fix the volume to 20mL.
3.8 Mixed standard working solution. Accurately weight 0.0500 g of caffeine that has
been dried at 80°C for 4h; add 5mL of standard stock solution A and 10mL of standard
stock solution B into the caffeine; then add mixed solution of methanol + water +
phosphoric acid =100 + 400 + 0.5 TO 20 mL.
4  Instruments 
4.1 High performance liquid chromatograph. attached with ultraviolet detector.
4.2 Acoustic cleaner.
4.3 Centrifuge.
5  Analysis procedures 
5.1 Sample treatment
5.1.1 Take more than 20 tablets or capsule samples; grind it or mix it evenly; weigh a
certain amount of sample (accurate to 0.001g) and place into a test tube; add methanol +
water + phosphate = 100 + 400 + 0.5 mixed solution; make the concentration to be – in
each 1mL, it contains thiamine hydrochloride, pyridoxine hydrochloride 0.25 mg; and it
contains niacin, niacinamide 1 mg. After the supersonic extraction for 5 min, centrifuge it
at 3000r/min for 5 minutes; filter the supernatant through 0.45μm membrane, it is ready for
injection.
5.1.2 Liquid sample is directly diluted with methanol + water + phosphate = 100 + 400
+ 0.5 mixed solution; mixed evenly; filter through 0.45 μm membrane, it is ready for
......
 
(Above excerpt was released on 2016-08-26, modified on 2021-06-07, translated/reviewed by: Wayne Zheng et al.)
Source: https://www.chinesestandard.net/PDF.aspx/GBT5009.197-2003