HPLC Report page 1 of 4

HPLC Chromatogram Result, Sample name: Metformin-HCl; Phenytoin Standard

Chr&HPLC,&Sample&name:&Metformin<HCl;&Phenytoin&Standard

590

540

490
Metformin9HCl) (2,5)µg/ml)
440
Metformin9HCl) (2)µg/ml)
390

mAU 340 Metformin9HCl) (1)µg/ml)

290 Metformin9HCl) (0,5)µg/ml)

Phenytoin)(2,5)µg/ml)
240 Metformin9HCl) (0,25)µg/ml)
Phenytoin)(2)µg/ml)
190 Metformin9HCl) (0,125)µg/ml)
Phenytoin)(1)µg/ml)
140 Phenytoin)(0,5)µg/ml)

90 Phenytoin)(0,25)µg/ml)
Phenytoin)(0,125)µg/ml)
40

910
0

10

11

12

13

14

15

16

17

18

19

20
7,561

9,929
RT& (min)

Table 1. HPLC Chromatogram Result, Standard concentration: 0,125 μg/ml

Standard concentration RT (min) Standard curve area Compound

(μg/ml)

0,125 7,561 6,12823 Phenytoin

9,929 5,17662 Metformin-HCl

 HPLC Report page 2 of 4

Table 2. HPLC Chromatogram Result, Standard concentration: 0,25 μg/ml

Standard concentration RT (min) Standard curve area Compound

(μg/ml)

0,25 7,561 9,74928 Phenytoin

9,929 17,91715 Metformin-HCl

Table 3. HPLC Chromatogram Result, Standard concentration: 0,5 μg/ml

Standard concentration RT (min) Standard curve area Compound

(μg/ml)

0,5 7,561 17,55249 Phenytoin

9,929 46,18273 Metformin-HCl

Table 4. HPLC Chromatogram Result, Standard concentration: 1 μg/ml

Standard concentration RT (min) Standard curve area Compound

(μg/ml)

1 7,561 30,92116 Phenytoin

9,929 97,52331 Metformin-HCl

Table 5. HPLC Chromatogram Result, Standard concentration: 2 μg/ml

Standard concentration RT (min) Standard curve area Compound

(μg/ml)

2 7,561 56,42338 Phenytoin

9,929 227,26983 Metformin-HCl

 HPLC Report page 3 of 4

Table 6. HPLC Chromatogram Result, Standard concentration: 2,5 μg/ml

Standard concentration RT (min) Standard curve area Compound

(μg/ml)

2,5 7,561 81,42152 Phenytoin

9,929 421,82736 Metformin-HCl

Table 7. Linear Regression Equation

Standard Linear Regression Equation r

Phenytoin y = 1,7519 + 30,0681x 0,9941

Metformin-HCl y = −33,8421+ 159,8353x 0,9693

Curve Area Ratio (Metformin-HCl/Phenytoin) y = 1,3403+ 1,5113x 0,9583

 HPLC Report page 4 of 4

HPLC Properties

HPLC apparatus model Shimadzu

System controller SCL 10 AVP

Solvent delivering unit LC 20AT

Column oven CTO 10 ASVP

Detector SPD 20-A UV Vis Detector

Column Shim-pack GIS-HP C 18 150 mmL x 3,0 mmID, 3 μm

Column temperature 25 °C

Mobile phase Acetonitrile 34% in KH2PO4 10 mM and Sodium lauryl sulfate 10 mM

Mobile phase method Isocratic

Flow rate 1,3 ml/min

Injection volume 20 μl

Wavelenght detector 233 nm

Run time 20 minutes

HPLC method Metformin method

User name Endang S, Unair, Surabaya

Sample name Metformin-HCl; Phenytoin Standard

Injection time 12/1/2016