Determination of Probiotic Abilities and Lactic Acid Content of Pediococcus acidilactici

2019 ◽  
Vol 15 (4) ◽  
pp. 511-521 ◽  
Author(s):  
Merve Eylul Kiymaci ◽  
Mehmet Gumustas ◽  
Nurten Altanlar ◽  
Ahmet Akin ◽  
Aysegul Zenciroglu ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
High Performance Liquid Chromatography ◽  
Lactic Acid ◽  
Liquid Chromatography ◽  
Acid Content ◽  
High Performance ◽  
Low Ph ◽  
Pediococcus Acidilactici ◽  
Lactic Acid Content

Background:Probiotics are living microorganisms that have a healthy influence on a host.Objective:The aim of this study was to isolate a probiotic Pediococcus acidilactici strain from newborn faeces and develop and optimize a selective high-performance liquid chromatography method for the determination and validation of its lactic acid content and also evaluate some probiotic characteristics.Methods:Isolated strains were identified by the API 50 CH system and 16S rDNA gene sequence analysis and tested for antibiotic susceptibility, bile salt tolerance, low pH resistance, proteolytic, haemolytic activity, as well as the production of bacteriocin, hydrogen peroxide, and lactic acid. Antimicrobial activity of selected strain against standard test microorganisms was determined by the spot lawn method and the quantitation of lactic acid was carried out by high-performance liquid chromatography on a Rezex ROA organic acid (300x7.8 mm) analytical column.Results:P. acidilactici M7 strain was evaluated as a potential probiotic due to its ability to survive at low pH values or in the presence of pepsin, pancreatin, and bile salts. The lactic acid amount of strain was found in the range between 5.59-5.94 mg mL-1 by HPLC. M7 strain was also found to be resistant to vancomycin, had no bacteriocin, and hydrogen peroxide production and was able to inhibit the growth of P. aeruginosa and E. faecalis by its lactic acid content.Conclusion:This study explains a simple, selective, and fully validated procedure for the determination of lactic acid from probiotic bacteria.

scholarly journals Optimasi dan Validasi Metode Penentuan Kadar Asam Glikolat dan Asam Laktat Dalam Krim Menggunakan Kromatografi Cair Kinerja Tinggi

2020 ◽  
Vol 16 (1) ◽  
pp. 10
Author(s):  
Devi Wulandari ◽  
Gusrizal Gusrizal ◽  
Titin Anita Zaharah
Keyword(s):  
High Performance Liquid Chromatography ◽  
Lactic Acid ◽  
Liquid Chromatography ◽  
High Performance ◽  
Glycolic Acid ◽  
Orthophosphoric Acid ◽  
Spiked Sample ◽  
Relative Standard ◽  
Uv Visible

<p>Telah dilakukan optimasi dan validasi metode penentuan asam glikolat dan asam laktat dalam krim menggunakan kromatografi cair kinerja tinggi. Pemisahan asam glikolat dan asam laktat dilakukan pada kolom fasa balik C-8menggunakan fasa gerak asam ortofosfat 0,1% dengan pH 3,5 dan detektor <em>UV-Visible</em>. Standar asam glikolat dan asam laktat dibuat dengan melarutkannya menggunakan asam ortofosfat 0,1% pH 3,5. Hasil eksperimen menunjukkan bahwa pada rentang konsentrasi 25 – 400 μg/mL, asam glikolat dan asam laktat memiliki kurva yang linear dengan koefisien korelasi masing-masing 0,9997 dan 0,9999. Uji presisi untuk larutan standar berkonsentrasi 100 µg/mL menghasilkan simpangan baku relatif sebesar 1,49% untuk asam glikolat dan 1,72% untuk asam laktat. Metode yang telah dioptimasi memberikan akurasi yang baik yang ditunjukkan oleh nilai perolehan kembali dari pengukuran tiga <em>spiked sample</em> dengan konsentrasi berbeda (50, 100, dan 150 μg/mL). Nilai perolehan kembali untuk masing-masing konsentrasi <em>spiked sample</em> adalah 97,12% ± 0,69; 98,76% ± 0,43; 100,80% ± 0,29 untuk asam glikolat dan 97,58% ± 0,39; 96,20% ± 0,68; 98,00% ± 0,38 untuk asam laktat. Batas deteksi dan batas kuantisasi untuk asam glikolat adalah 0,05 dan 0,17 μg/mL, sedangkan untuk asam laktat adalah 1,40 dan 4,67 μg/mL. Nilai kekasaran metode untuk asam glikolat pada hari pertama dan hari kedua adalah 1,43% dan 1,67%, sedangkan untuk asam laktat adalah 1,67% dan 1,25%. Metode yang telah dioptimasi dan divalidasi berpotensi untuk digunakan secara spesifik pada penentuan kadar asam glikolat dan asam laktat dalam krim.</p><p><strong>Optimization and Validation of Determination Methods of Glycolic Acid and Lactic Acid in Cream Using High-Performance Liquid Chromatography.</strong> A high-performance liquid chromatography analytical method for the determination of glycolic acid and lactic acid in creams has been optimized and validated. The separation was performed in a reverse phase C–8 column with a mobile phase of 0.1%, orthophosphoric acid at pH 3.5, and UV-Visible detector. The standard of glycolic acid and lactic acid was dissolved in 0.1% orthophosphoric acid at pH 3.5. The experimental results showed that in the concentration range of 25–400 μg/mL, glycolic acid and lactic acid showed a linear curve with a correlation coefficient of 0.9997 and 0.9999, respectively. The precision test for standard solutions containing 100 µg/mL resulted in a relative standard deviation of 1.49% for glycolic acid and 1.72% for lactic acid. The optimized method provided good accuracy indicated by the recovery of the measurement of three spiked samples in different concentrations (50, 100, and 150 μg/mL). The recovery for each concentration of the spiked sample was 97.12% ± 0.69; 98.76% ± 0.43; 100.80% ± 0.29 for glycolic acid and 97.58% ± 0.39; 96.20% ± 0.68; 98.00% ± 0.38 for lactic acid. The limit of detection and limit of quantization for glycolic acid was 0.05 and 0.17 μg/mL, and for lactic acid was 1.40 and 4.67 μg/mL. The ruggedness of the method for glycolic acid on the first day and second day was 1.43% and 1.67%, while for lactic acid, it was 1.67% and 1.25%. The method that has been optimized and validated shows the potential to be used specifically for the determination of glycolic acid and lactic acid in the cream.</p>

scholarly journals Simultaneous Determination of Kynurenine and Kynurenic Acid by High-Performance Liquid Chromatography Photoirradiation System Using a Mobile Phase Containing 18-crown-6

2019 ◽  
Vol 12 ◽  
pp. 117864691983455
Author(s):  
Motomasa Atsumi ◽  
Ken-ichi Mawatari ◽  
Akari Morooka ◽  
Makoto Yasuda ◽  
Tomoko Fukuuchi ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Phosphate Buffer ◽  
Mobile Phase ◽  
High Performance ◽  
Kynurenic Acid ◽  
Fluorometric Determination ◽  
The Mean

A high-performance liquid chromatography (HPLC) system has been developed for the fluorometric determination of kynurenine (KYN) and kynurenic acid (KYNA) in human serum using a mobile phase containing 18-crown-6. A retention time of KYNA was adjusted with pH of phosphate buffer in 18-crown-6. KYN and KYNA were separated on a CAPCELLPAK C18 (250 × 4.6 mm i.d.). The mobile phase consisted of 35 mmol/L phosphate buffer (pH 8.0)/methanol (85/15, v/v) containing 35 mmol/L hydrogen peroxide and 10 mmol/L 18-crown-6. The retention times of KYN and KYNA were 18and 24 minutes, respectively. The calibration graphs of KYN and KYNA were linear over the range 180 to 2900 and 1 to 84 nmol/L by injecting a 50-μL volume of KYN and KYNA, respectively. Pretreatment of serum was achieved by deproteinization only. The mean recoveries of KYN and KYNA from serum were more than 97%.

Method for determination of hydrogen peroxide in adulterated milk using high performance liquid chromatography

2019 ◽  
Vol 283 ◽  
pp. 431-436 ◽  
Author(s):  
Anastasia S. Ivanova ◽  
Anna D. Merkuleva ◽  
Sergey V. Andreev ◽  
Konstantin A. Sakharov
Keyword(s):  
Hydrogen Peroxide ◽  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance
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