A novel RP-HPLC method for simultaneous determination of vitamins B1, B2, B3, B6 and C in oral powder for veterinary consumption

Ahmet Haşimoğlu

Article Details

A novel RP-HPLC method for simultaneous determination of vitamins B1, B2, B3, B6 and C in oral powder for veterinary consumption

Authors:

Ahmet Haşimoğlu

Publisher:

Marmara University

Main Topics:

Pharmaceutical Analytical Chemistry

Journal:

Journal of Research in Pharmacy Vol.22 Issue 4, pp.484-492

Document Type:

Article

Publication Date:

6/2025

Document Language:

English

Document ID (Org No.):

2001346451

pISSN:

2630-6344

Keywords:

Abstract:

 The water-soluble vitamins are gaining rapid importance not only for human health but also being widely recommended for animal health care because of theirs wide spectrum of activities ranging from deficiency, pathogenic condition to nutritional supplements. A plethora of articles are published for simultaneous evaluation of B-Complex Vitamins, but most of their methods are not selective and time consuming which involve pre-treatment of sample due to complexities involved in the placebo matrix. In this study, we propose a simple, specific, precise, accurate and linear method to separate vitamin B1 thiamine, vitamin B2 riboflavin sodium phosphate, vitamin B3 niacin amide, vitamin B6 pyridoxine and vitamin C ascorbic acid by using Zorbax XDB C18 column 250 mm x 4.6 mm i.d., 5µm. The flow rate was kept as 1.3 mLmin and monitored at 270 nm with ambient temperature. Effective separations and quantification were obtained in 30 minutes with 10 µl injection volume. The current reverse phase gradient method resulted in significant resolution between interested vitamins and placebo matrix. Analytical method development, followed by analytical method validation was accomplished according to International Conference of Harmonization guidelines ICH. Statistical evaluation of validation parameters proved that the results were highly specific, precise, accurate, robust and linear over a wide concentration range. This method has been validated and found to be applicable in routine analysis of vitamin B1, vitamin B2, vitamin B3, vitamin B6 and vitamin C in oral powder for veterinary consumption.

 The water-soluble vitamins are gaining rapid importance not only for human health but also being widely recommended for animal health care because of theirs wide spectrum of activities ranging from deficiency, pathogenic condition to nutritional supplements. A plethora of articles are published for simultaneous evaluation of B-Complex Vitamins, but most of their methods are not selective and time consuming which involve pre-treatment of sample due to complexities involved in the placebo matrix. In this study, we propose a simple, specific, precise, accurate and linear method to separate vitamin B1 thiamine, vitamin B2 riboflavin sodium phosphate, vitamin B3 niacin amide, vitamin B6 pyridoxine and vitamin C ascorbic acid by using Zorbax XDB C18 column 250 mm x 4.6 mm i.d., 5µm. The flow rate was kept as 1.3 mLmin and monitored at 270 nm with ambient temperature. Effective separations and quantification were obtained in 30 minutes with 10 µl injection volume. The current reverse phase gradient method resulted in significant resolution between interested vitamins and placebo matrix. Analytical method development, followed by analytical method validation was accomplished according to International Conference of Harmonization guidelines ICH. Statistical evaluation of validation parameters proved that the results were highly specific, precise, accurate, robust and linear over a wide concentration range. This method has been validated and found to be applicable in routine analysis of vitamin B1, vitamin B2, vitamin B3, vitamin B6 and vitamin C in oral powder for veterinary consumption.

 The water-soluble vitamins are gaining rapid importance not only for human health but also being widely recommended for animal health care because of theirs wide spectrum of activities ranging from deficiency, pathogenic condition to nutritional supplements. A plethora of articles are published for simultaneous evaluation of B-Complex Vitamins, but most of their methods are not selective and time consuming which involve pre-treatment of sample due to complexities involved in the placebo matrix. In this study, we propose a simple, specific, precise, accurate and linear method to separate vitamin B1 thiamine, vitamin B2 riboflavin sodium phosphate, vitamin B3 niacin amide, vitamin B6 pyridoxine and vitamin C ascorbic acid by using Zorbax XDB C18 column 250 mm x 4.6 mm i.d., 5µm. The flow rate was kept as 1.3 mLmin and monitored at 270 nm with ambient temperature. Effective separations and quantification were obtained in 30 minutes with 10 µl injection volume. The current reverse phase gradient method resulted in significant resolution between interested vitamins and placebo matrix. Analytical method development, followed by analytical method validation was accomplished according to International Conference of Harmonization guidelines ICH. Statistical evaluation of validation parameters proved that the results were highly specific, precise, accurate, robust and linear over a wide concentration range. This method has been validated and found to be applicable in routine analysis of vitamin B1, vitamin B2, vitamin B3, vitamin B6 and vitamin C in oral powder for veterinary consumption.

 The water-soluble vitamins are gaining rapid importance not only for human health but also being widely recommended for animal health care because of theirs wide spectrum of activities ranging from deficiency, pathogenic condition to nutritional supplements. A plethora of articles are published for simultaneous evaluation of B-Complex Vitamins, but most of their methods are not selective and time consuming which involve pre-treatment of sample due to complexities involved in the placebo matrix. In this study, we propose a simple, specific, precise, accurate and linear method to separate vitamin B1 thiamine, vitamin B2 riboflavin sodium phosphate, vitamin B3 niacin amide, vitamin B6 pyridoxine and vitamin C ascorbic acid by using Zorbax XDB C18 column 250 mm x 4.6 mm i.d., 5µm. The flow rate was kept as 1.3 mLmin and monitored at 270 nm with ambient temperature. Effective separations and quantification were obtained in 30 minutes with 10 µl injection volume. The current reverse phase gradient method resulted in significant resolution between interested vitamins and placebo matrix. Analytical method development, followed by analytical method validation was accomplished according to International Conference of Harmonization guidelines ICH. Statistical evaluation of validation parameters proved that the results were highly specific, precise, accurate, robust and linear over a wide concentration range. This method has been validated and found to be applicable in routine analysis of vitamin B1, vitamin B2, vitamin B3, vitamin B6 and vitamin C in oral powder for veterinary consumption.

Article Details

A novel RP-HPLC method for simultaneous determination of vitamins B1, B2, B3, B6 and C in oral powder for veterinary consumption

Authors:

Ahmet Haşimoğlu

Publisher:

Marmara University

Main Topics:

Pharmaceutical Analytical Chemistry

Journal:

Journal of Research in Pharmacy Vol.22 Issue 4, pp.484-492

Document Type:

Article

Publication Date:

6/2025

Document Language:

English

Document ID (Org No.):

2001346451

pISSN:

2630-6344

RP-HPLC

simultaneous determination

vitamin C, B1, B2, B3, B6

water-soluble vitamins

veterinary

Issues

Article Details

A novel RP-HPLC method for simultaneous determination of vitamins B1, B2, B3, B6 and C in oral powder for veterinary consumption

Authors: Ahmet Haşimoğlu

Publisher: Marmara University

Main Topics: Pharmaceutical Analytical Chemistry

Journal: Journal of Research in Pharmacy Vol.22 Issue 4, pp.484-492

Document Type: Article

Publication Date: 6/2025

Document Language: English

Document ID (Org No.): 2001346451

pISSN: 2630-6344

RP-HPLC

simultaneous determination

vitamin C, B1, B2, B3, B6

water-soluble vitamins

veterinary

Issues

Authors:

Ahmet Haşimoğlu

Publisher:

Marmara University

Main Topics:

Pharmaceutical Analytical Chemistry

Journal:

Journal of Research in Pharmacy Vol.22 Issue 4, pp.484-492

Document Type:

Article

Publication Date:

6/2025

Document Language:

English

Document ID (Org No.):

2001346451

pISSN:

2630-6344