Method development for folate determination using local lactobacillus casei / by Faith Jennah L. Corpuz

Thesis (BS Medical Technology) Cavite State University

Includes bibliographical references.

College of Nursing (CON)

CORPUZ, FAITH JENNAH L., CORTEZ, PATRICIA P., PESTRANA, ROLAND G., SANGALANG, ANDREA MICAELA C. Method Development For Folate Determination Using Local Lactobacillus casei. Undergraduate Thesis. Bachelor of Science in Medical Technology, Cavite State University, Indang, Cavite. April 2016. Adviser: Dr. Hosea L. Matel.

Generally, this study aimed to develop a method for the quantitation of folate levels using local Lactobacillus casei. The study included modifications which was based from the original microbiological assay which was done by previous researchers. The modifications that were used in the study included: use of local strain of Lactobacillus case! which was isolated from fermented mung bean, lyophilized human serum controls and reading of results at 16, 24, 42 and 48 hours of incubation. The study was conducted from December 2015 to March 2016 in the College of Nursing and Research Center of Cavite State University, Indang, Cavite. Lyophilized human serum control with Folate, levels 1 (1.00 - 1.60 ng/mL) and 3 (11.0 - 16.0 ng/mL) was procured at the Bio-Rad Laboratories, 9500 Jeronimo Road, Irvine, California. Lactobacillus casei was procured at the National Institute of Molecular Biology and Biotechnology, University of the Philippines Los Barios, Laguna, Philippines. The procured microorganism was sub-cultured using de Man, Rogosa and Sharpe (MRS) Broth, incubated at 37°C aerobically for 48-72 hours. Personal Protective Equipment (PPE) and other miscellaneous materials that was used in the study was procured at Bambang, Sta. Cruz, Manila. The working standard and 0.5% Sodium Ascorbate were prepared. Microtiter plates with working standard, 0.5% Sodium Ascorbate, and Lactobacillus case! added was incubated aerobically at 37°C.

The microtiter plates were read at 546 nm using the Dynex Technologies Microtiter Plate Reader and the optical density readings at 16, 24, 42, and 48-hour incubation period were recorded. Computation of the actual concentration of each well was calculated using the known optical density of standards to get the Folic Acid concentration. From the data that was gathered from the reader, a linear regression graph was constructed from each plate with corresponding levels: Monday microtiter plates were read at Tuesday and Wednesday (10:00 AM & 4:00 PM) for levels 1 and 3. Tuesday microtiter plates were read at Wednesday and Thursday (10:00 AM & 4:00 PM) for levels 1 and 3. Wednesday microtiter plates were read at Thursday and Friday (10:00 AM & 4:00 PM) for levels 1 and 3. Thursday microtiter plates were read at Friday and Saturday (10:00 AM & 4:00 PM) for levels 1 and 3. Finally, Friday microtiter plates were read at Saturday and Sunday (10:00 AM & 4:00 PM) for levels 1 and 3. Linear regression graph readings for Figures 18, 19, 22, and 23 shows trend between the Folic Acid concentration and absorbance; as the Folic Acid concentration increases, the spectral bacterial growth also increases. Hence, there is a correlation between the absorbance at 546 nm in that particular reading which was incubated at 42, 48, 16 and 24 hours, respectively, to the Folic Acid concentration. But since only four set ups out of 40 shows this result, this method was proven to have a low degree of reproductivity in determining Folic Acid.

Submitted copy to the University Library. 12/06/2016 T-6084