Objective:

2

To establish an LC-MS/MS method for the determination of alcohol-soluble components and water-soluble components in Arnebiae Radix(

L

-shikonin

β

β

′-dimethylacrylshikonin

β

-acetoxy-isovalerylshikonin

lithospermic acid

caffeic acid

rosmarinic acid)

for the purpose of quality control of the herb.

Method:

2

Chromatographic separation was carried out at 35 ℃ on Agilent ZORBAX SB C

18

(4.6 mm×50 mm

1.8 μm)

with 0.1% formic acid acetonitrile (A)-0.1% formic acid solution (B) as the mobile phase for gradient elution (0-3 min

5%-95%A; 3-7 min

95%A; 7-7.01 min

95%-5%A; 7.01-8.5 min

5%A). The flow rate was 0.2 mL·min

-1

and the injection volume was 5 μL. In a negative ionization mode

MRM scanning mode was adopted for quantification.

Result:

2

The calibration curves of

L

-shikonin

β

β

′-dimethylacrylshikonin

β

-acetoxy-isovalerylshikonin

lithospermic acid

caffeic acid

rosmarinic acid showed a good linearity

and the linear ranges of the above six compounds were 10.12-1 012 μg·L

-1

(

r

=0.998 1)

10.88-1 088 μg·L

-1

(

r

=0.991 2)

10.08-806.4 μg·L

-1

(

r

=0.997 6)

20.32-1 016 μg·L

-1

(

r

=0.996 6)

10.37-1 037 μg·L

-1

(

r

=0.999 6)

10.26-1 026 μg·L

-1

(

r

=0.997 8)

respectively. The average recoveries of the analysts were 95.8%(RSD 3.2%)

103.5%(RSD 2.3%)

105.3%(RSD 2.1%)

96.1%(RSD 3.3%)

98.9%(RSD 2.7%)

100.8%(RSD 3.4%). The contents of six components in 13 batches of samples showed significant differences.

Conclusion:

2

The established method is feasible and simple

and provides a basis for comprehensive quality evaluation of Arnebiae Radix.