This document describes a method for analyzing formic acid using high performance liquid chromatography (HPLC). The method involves using a C18 column with a mobile phase of potassium phosphate buffer at pH 2.5. Standards were prepared with concentrations ranging from 10-1200 ppm formic acid. The results show a linear relationship between concentration and peak area with an R2 value of 0.9911. The method was then used to analyze formic acid levels in Biotronic products, showing consistent recovery when samples were spiked with additional formic acid.

1. Formic acid analysis method
standardized
using HPLC
Karthikeyan

2. • Aim
• To estimate amount of formic acid present in product and competitor
product by using HPLC
• Supporting to salcurb asf and myco curb products
FORMIC ACID
• Formic acid is a preservative and antimicrobial agent in animal feeds, when
spray on feed and silage, it's retain long nutritive value

3. HPLC Condition
• Column:5 µm C18 125 Å, LC Column 250 x 4.6 mm, Ea
• Mobile Phase ;25um KH2 Po4 Buffer solution
• Elution Type:Isocratic
• Wave length:220
• Flow rate:0.6ml/per min

4. Formic acid standard recovery
10 25 50 75 107.3 1204.27 4.251 4.24 4.238 4.244 4.244
98950
243750.8
492772.4
718871.6
1041682.275
1250019
y = 239293x - 196516
R² = 0.9911
1 2 3 4 5 6
Formic acid standard recovery
PPM RT Area
100 4.27 98950
250 4.251 243750.8
500 4.24 492772.4
750 4.238 718871.6
1070.3 4.244 1041682
1200 4.244 1250019

5. Formic acid standard Biotronic product result
y = -29205x + 186904
R² = 0.9908
0
20000
40000
60000
80000
100000
120000
140000
160000
180000
1 2 3 4 5 6
48.8 mg sample+ 24.4 mgstd spike recovery
S.No 48.8 mg sample and std pike
50 4.507 161639
40 4.512 130386
30 4.512 89913.4
20 4.512 71887.9
10 4.512 38128.5
5 4.5 16163.9

6. Formic acid standard Biotronic product result
S.No 98.4 mg sample and std pike
100 4.6 194247.5
80 4.6 156308.5
60 4.6 118741.7
40 4.6 81169.8
20 4.6 36353.4
10 4.6 19876.1
y = -36266x + 228046
R² = 0.9911
0
50000
100000
150000
200000
250000
1 2 3 4 5 6
98.8mg sample +22.4mg sample spike

7. Formic acid standard Biotronic product result
4.6 4.6 4.6 4.6 4.6 4.6
230344.75
195518.2
147152.2
108143.5
70193.1
12789.5
y = -42936x + 277633
R² = 0.9955
0
50000
100000
150000
200000
250000
1 2 3 4 5 6
148.9mg sample+ 25.4mg std S.No 148.9 mg sample and std pike
150 4.6 230344.75
130 4.6 195518.2
100 4.6 147152.2
75 4.6 108143.5
50 4.6 70193.1
25 4.6 12789.5

8. Chromatogram Of Formic acid