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A HPLC-UV Method for Simultaneous Determination of Six Steroid Hormone Drugs

Method: HPLC
Matrix: Medicine
Application No.: 101956
Componds: Hydrocortisone, Estradiol, Equilin, Estrone, Methyltestosterone, Stanozolol
Columns: Diamonsil® Plus C18 5μm 250 x 4.6 mm
Cat No.: 99403
Sample Pretreatment: Standards: Individual stock solutions were prepared in acetonitrile at a concentration of 1.0 mg/mL. A working solution of the mixture of all 6 drugs was obtained by dilution of the stock solution with acetonitrile/water (20/80).
Conditions: Mobile Phase:
A: 30mM Potassium Phosphate Dibasic (pH 8.0);
B: Acetonitrile: Methanol = 4 / 6
Gradient: hold 40% B for 1 min, 40 to 60% B in 3 min, hold 4 min, 60 to 70% B in 4 min, hold 6 min, 70 to 40% B in 1 min, equilibrate at 40% B for 6 min.
Flow Rate: 1.0 mL/min
Injection: 10 µL
Temperature: 30 ℃
Wavelength: 230 nm
Publisher: Dikma Technologies Inc.
Keyword: Hydrocortisone, Estradiol, Equilin, Estrone, Methyltestosterone, Stanozolol, Diamonsil Plus C18
Abstract: All analytes were successfully eluted from DIKMA Diamonsil Plus C18 column within 19 min. The proposed method is simple, efficient, and can be useful and reliable for quality control analysis in SHDs research and production.

 

No.

Compounds

Concentration 

(µg/mL)

Retention Time

(min)

Resolution

Asymmetry

1

Hydrocortisone

10

8.807

-

1.09

2

Estradiol

20

12.480

18.92

1.06

4

Equilin

20

13.071

2.56

1.07

5

Estrone

20

13.650

2.59

1.06

5

Methyltestosterone

20

15.617

8.95

1.04

6

Stanozolol

40

18.576

11.55

1.10