Thiamine Injection

Vitamin B₁.

Thiamine Injection is a sterile solution of Thiamine Hydrochloride in Water for Injections.

The injection complies with the requirements stated under Parenteral Preparations and with the following requirements.

95.0  to 105.0% of the stated amount.

A colourless or almost colourless solution.

A.  Carry out the method for thin-layer chromatography, Appendix III A, using cellulose F₂₅₄ as the coating substance and a mixture of 60  volumes of butan-1-ol, 25  volumes of water and 15  volumes of glacial acetic acid as the mobile phase. Apply separately to the plate 2 µl of each of the following solutions. For solution (1) use the injection diluted if necessary with water to contain 0.1% w/v of Thiamine Hydrochloride. Solution (2) contains 0.1% w/v of thiamine mononitrate BPCRS in water. After removal of the plate, allow it to dry in air, heat at 105° for 30  minutes, spray with a mixture of equal volumes of a 0.3% w/v solution of potassium hexacyanoferrate(III) and a 10% w/v solution of sodium hydroxide and examine under ultraviolet light (365  nm). The principal spot in the chromatogram obtained with solution (1) corresponds to that in the chromatogram obtained with solution (2).

B.  To a volume containing 20  mg of Thiamine Hydrochloride diluted, if necessary, to 10  ml with water, add 2  ml of 1m acetic acid and 1.6  ml of 1m sodium hydroxide, heat in a water bath for 30  minutes and cool. Add 5  ml of 5m sodium hydroxide, 10  ml of dilute potassium hexacyano-ferrate(III) solution and 10  ml of butan-1-ol and shake vigorously for 2  minutes. The upper layer shows an intense light blue fluorescence on exposure to ultraviolet light. Repeat the test but adding 0.9  ml of 1m sodium hydroxide and 0.2  g of sodium sulphite in place of the 1.6  ml of 1m sodium hydroxide. Not more than a slight fluorescence is produced.

C.  To a mixture of 0.1  ml of nitrobenzene and 0.2  ml of sulphuric acid add a volume of the injection containing 5  mg of Thiamine Hydrochloride. Allow to stand for 10  minutes, cool in ice and add slowly with stirring 5  ml of water followed by 5  ml of 10m sodium hydroxide. Add 5  ml of acetone and allow to stand. No violet colour is produced in the upper layer.

pH, 2.8  to 3.4, Appendix V L.

Carry out the method for liquid chromatography, Appendix III D, using the following solutions. Solution (1) contains 0.005% w/v of thiamine mononitrate BPCRS in 0.005m hydrochloric acid. For solution (2) dilute a volume of the injection containing 0.1  g of Thiamine Hydrochloride to 100  ml with 0.1m hydrochloric acid and further dilute 5  ml to 100  ml with water.

The chromatographic procedure may be carried out using (a) a stainless steel column (10  cm × 4.6  mm) packed with end-capped octadecylsilyl silica gel for chromatography (5 µm) (Nucleosil C18 is suitable), (b) as the mobile phase with a flow rate of 2  ml per minute a solution prepared by dissolving 1  g of sodium heptane-sulphonate in a mixture of 180  ml of methanol and 10  ml of triethylamine, diluting to 1000  ml with water and adjusting the pH to 3.2 with orthophosphoric acid and (c) a detection wavelength of 244  nm.

Calculate the content of C₁₂H₁₇ClN₄OS,HCl using the declared content of C₁₂H₁₇N₅O₄S in thiamine mononitrate BPCRS. Each mg of C₁₂H₁₇N₅O₄S is equivalent to 1.030  mg of C₁₂H₁₇ClN₄OS,HCl.

Thiamine Injection should be protected from light.

When vitamin B₁ injection is prescribed or demanded, Thiamine Injection shall be dispensed or supplied.