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UHPLC/MS/MS Method for Simultaneous Determination of 5Benzodiazepines

Method: HPLC-MS
Matrix: Medicine
Application No.: 101930
Componds: Chlordiazepoxide, Clonazepam, Oxazepam, Alprazolam, Diazepam
Columns: Endeavorsil C18 1.8μm , 150x2.1mm
Cat No.: 87004
Sample Pretreatment: Standards: Individual stock solutions were prepared in acetonitrileat a concentration of 0.5 mg/mL. A working solution of the mixture of all 5 drugs was obtained by dilution of the stock solution with mobile phase. All the stock solutions should be stored at -20 ℃. Working solution are prepared when needed and used out as soon as possible.
Conditions: Mobile Phase:
A: 10 mM ammonium formate, pH 3.0
B: methanol:acetonitrile = 4:6
Gradient: hold 40% B for 1.5 min, 40 to 60% B in 0.5 min, 60 to 65% B in 5.5 min, 65 to 70% B in 0.5 min, hold 70% B for 2 min, 70 to 40% B in 0.5 min, equilibrate at 40% B for 4.5 min.
Flow Rate: 0.2 mL/min;
Injection: 2 µL;
Temperature: 25 0C
Source: ESI; Ionization Mode: Positive
Instrument: Agilent 1290 Infinity System and AB Sciex API 4000+
Publisher: Dikma Technologies Inc.
Keyword: Chlordiazepoxide, Clonazepam, Oxazepam, Alprazolam, Diazepam, UHPLC/MS/MS, 5Benzodiazepines
Abstract: All analytes were eluted from the column within 10 mins with good peak shape and resolution. Test results showed that it is a rapid, simple method and can be a good recommendation for routine lab work.

 

No.

Compounds

Concentration (µg/mL)

Retention Time (min)

MS→MS1 Ions

(Precursor→Product 1)

MS→MS2 Ions

(Precursor→Product 2)

1

Chlordiazepoxide

0.15

4.77

300.1→227.0

300.1→283.1

2

Clonazepam

0.40

6.13

316.1→270.1

316.1→214.2

3

Oxazepam

0.16

6.37

287.1→269.0

287.1→241.0

4

Alprazolam

0.05

6.82

309.1→280.9

309.1→205.2

5

Diazepam

0.10

9.39

285.1→154.0

285.1→193.0