Method development for the analysis of beta-carotene and astaxanthin by capillary electrophoresis / Khin Thida Nyunt
Issued Date
2007
Copyright Date
2007
Resource Type
Language
eng
File Type
application/pdf
No. of Pages/File Size
xiv, 120 leaves : ill.
Access Rights
open access
Rights
ผลงานนี้เป็นลิขสิทธิ์ของมหาวิทยาลัยมหิดล ขอสงวนไว้สำหรับเพื่อการศึกษาเท่านั้น ต้องอ้างอิงแหล่งที่มา ห้ามดัดแปลงเนื้อหา และห้ามนำไปใช้เพื่อการค้า
Rights Holder(s)
Mahidol University
Bibliographic Citation
Thesis (M.Sc. (Pharmaceutical Chemistry))--Mahidol University, 2007
Suggested Citation
Nyunt, Khin Thida, 1975- Method development for the analysis of beta-carotene and astaxanthin by capillary electrophoresis / Khin Thida Nyunt. Thesis (M.Sc. (Pharmaceutical Chemistry))--Mahidol University, 2007. Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/97097
Title
Method development for the analysis of beta-carotene and astaxanthin by capillary electrophoresis / Khin Thida Nyunt
Author(s)
Advisor(s)
Abstract
Oil-in-water (o/w) and water-in-oil (w/o) MEEKC were investigated for the separation of beta-cartoene and astaxanthin. Due to the instability of the carotenoids in acid pH (2.5), o/w MEEKC at acid pH was not suitable. O/W microemulsion with basic pH (9.2) gave poor separation and sensitivity for both carotenoids because of highly hydrophobic nature and solubility problems. The second approach, w/o MEEKC offered complete resolution of both carotenoids due to its unique separation mechanism. Development of w/o MEEKC optimum condition was investigated by varying injection time, oil and surfactant types, surfactant and water compositions, additional oils and capillary length. The w/o microemulsion buffer containing 9% (w/w) SDS, 80% (w/w) 1- butanol, 11% (w/w) 70 mM sodium acetate (pH 8), using temperature of 25 °C, the separating voltage of -30 kV and the total capillary length of 32 cm (effective length 23.5 cm) was optimized. Detection was by a diode array detector at 475 nm with 40 nm bandwidth. Both carotenoids could be resolved within 9 min with a resolution of 4.9. Method linearity was good with r2 of 0.997 for B-carotene and r2 of 0.996 for astaxanthin over the concentration of 20-120 µg/ml. The method precision was excellent with % RSD of 3.1 % for migration time, 3.8 % for peak area of B -carotene and 1.1% for migration time, 3.4 % peak area for astaxanthin. Limits of detections were 3.5 and 4 µg/ml (%RSD= 5.3%) and limit of quantitations were 11.5 and 14 µg/ml (%RSD = 5.8 %) for beta-carotene and astaxanthin, respectively.
Oil-in-water (o/w) and water-in-oil (w/o) MEEKC were investigated for the separation of beta-cartoene and astaxanthin. Due to the instability of the carotenoids in acid pH (2.5), o/w MEEKC at acid pH was not suitable. O/W microemulsion with basic pH (9.2) gave poor separation and sensitivity for both carotenoids because of highly hydrophobic nature and solubility problems. The second approach, w/o MEEKC offered complete resolution of both carotenoids due to its unique separation mechanism. Development of w/o MEEKC optimum condition was investigated by varying injection time, oil and surfactant types, surfactant and water compositions, additional oils and capillary length. The w/o microemulsion buffer containing 9% (w/w) SDS, 80% (w/w) 1- butanol, 11% (w/w) 70 mM sodium acetate (pH 8), using temperature of 25 °C, the separating voltage of -30 kV and the total capillary length of 32 cm (effective length 23.5 cm) was optimized. Detection was by a diode array detector at 475 nm with 40 nm bandwidth. Both carotenoids could be resolved within 9 min with a resolution of 4.9. Method linearity was good with r2 of 0.997 for B-carotene and r2 of 0.996 for astaxanthin over the concentration of 20-120 µg/ml. The method precision was excellent with % RSD of 3.1 % for migration time, 3.8 % for peak area of B -carotene and 1.1% for migration time, 3.4 % peak area for astaxanthin. Limits of detections were 3.5 and 4 µg/ml (%RSD= 5.3%) and limit of quantitation's were 11.5 and 14 µg/ml (%RSD = 5.8 %) for beta-carotene and astaxanthin, respectively.
Oil-in-water (o/w) and water-in-oil (w/o) MEEKC were investigated for the separation of beta-cartoene and astaxanthin. Due to the instability of the carotenoids in acid pH (2.5), o/w MEEKC at acid pH was not suitable. O/W microemulsion with basic pH (9.2) gave poor separation and sensitivity for both carotenoids because of highly hydrophobic nature and solubility problems. The second approach, w/o MEEKC offered complete resolution of both carotenoids due to its unique separation mechanism. Development of w/o MEEKC optimum condition was investigated by varying injection time, oil and surfactant types, surfactant and water compositions, additional oils and capillary length. The w/o microemulsion buffer containing 9% (w/w) SDS, 80% (w/w) 1- butanol, 11% (w/w) 70 mM sodium acetate (pH 8), using temperature of 25 °C, the separating voltage of -30 kV and the total capillary length of 32 cm (effective length 23.5 cm) was optimized. Detection was by a diode array detector at 475 nm with 40 nm bandwidth. Both carotenoids could be resolved within 9 min with a resolution of 4.9. Method linearity was good with r2 of 0.997 for B-carotene and r2 of 0.996 for astaxanthin over the concentration of 20-120 µg/ml. The method precision was excellent with % RSD of 3.1 % for migration time, 3.8 % for peak area of B -carotene and 1.1% for migration time, 3.4 % peak area for astaxanthin. Limits of detections were 3.5 and 4 µg/ml (%RSD= 5.3%) and limit of quantitation's were 11.5 and 14 µg/ml (%RSD = 5.8 %) for beta-carotene and astaxanthin, respectively.
Description
Pharmaceutical Chemistry (Mahidol University 2007)
Degree Name
Master of Science
Degree Level
Master's degree
Degree Department
Faculty of Pharmacy
Degree Discipline
Pharmaceutical Chemistry
Degree Grantor(s)
Mahidol University