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GB/T 5009.197-2003Determination of thiamine hydrochloride, pyridoxine hydrochloride niacin, niacinamide and caffeine in health foods (English PDF)

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Issued by

Ministry of Health of the People's Republic of China

Level / Type

National · Recommended

Issue date

August 11, 2003

Implementation date

January 1, 2004

Scope

GB/T 5009.197-2003 (Determination of thiamine hydrochloride, pyridoxine hydrochloride niacin, niacinamide and caffeine in health foods) is available as an English-translated PDF.

GB/T 5009.197-2003 — 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.

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Document preview — GB/T 5009.197-2003

National Standard of the People's Republic of China

ICS: 67.040
Classification: C 53

Issued by: Ministry of Health of the People's Republic of China

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

Foreword

This Standard was proposed by and shall be under the jurisdiction of the Ministry of Health

of the People's Republic of China.

Chief drafting organization of this Standard. Institute of Food Safety Control and

Inspection Ministry of Public Health.

Chief drafters of this Standard. Yang Dajin, Fang Congrong and Wang Zhutian.

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.

3.7 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.

4 Instruments

4.1 High performance liquid chromatograph. attached with ultraviolet detector.

4.2 Acoustic cleaner.

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.

5.2 Chromatographic condition

5.2.1 Chromatographic column. TSK C18 4.6mm×150mm, 5µm.

5.2.2 Column temperature. room temperature.

5.2.3 Detection wavelength. For thiamine hydrochloride, 260 nm. For caffeine, niacin,

nicotinamide, pyridoxine hydrochloride, 280nm.

5.2.5 Flow rate. 1mL/min.

5.2.6 Injection amount. 10µL.

5.3 Chromatogram

6 Result calculation

Reserve two significant digits for the calculation results.

7 Precision

The absolute difference of two independent determination results, under the repeatability

condition, shall not be over 10% of the arithmetic mean.

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This preview omits tables, figures, formulas and parts of the technical clauses. The complete document — 8 pages — is available in the English PDF.

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