Normaidah

K 100110129
 Ephedra sinica Stapf
• In TCM, it has been used medicinally as
a diaphoretic, stimulant, and
antiasthmatic
• Ephedra species are a result of their
contents of ephedrine alkaloids: E, NE,
PE, NPE, ME, and MPE. The
derivatization of alkaloids enhances the
sensitivity and allows for the chiral HPLC
separation of enantiomeric mixtures
• 2,3,5,6-tetramethylpyrazine (TMP)
Ma-Huang (volatile oil-alkaloid), in Ma-Huang it
plays a very important role in relieving
TCM asthma
The goal of the analytical experiments
• To find out if direct GC-MS can give good
separation and determination of ephedrine
alkaloids and tetramethylpyrazine in Ephedra
sinica Stapf
Active marker or analytical marker
chosen, and why ?

(1S,2S)-(+)-Norpseudoephedrine

2,3,5,6-Tetramethylpyrazine

ACTIVE MARKER
 Hewlett-Packard

Analytical Method
Method : GC-MS
Detektor : MSD
Column : HP-5 MS capillary column (30.0 m × 250
µm × 0.25 µm, 5% phenyl methyl siloxane)
Temp Col : 600C – 1000C at 250C/min, 1100C at
0.50C/min, 1200C at 1.00C/min
Temp Inj : 2800C (1 µL)
Temp MS : 2500C – EI with energy 70eV
Mobile Ph : He was 1.0 mL/min
Mode : Both the full-scan acquisition mode (m/z 35-400)
Selected ion monitoring (SIM) (m/z 58,44,136)
 Preparation sample
Two grams of raw medicinal material of Ephedra sinica Stapf was
crushed into small pieces

Concentrated NH4OH (5mL) was added, the mixture was extracted with
60mL ethanol–ethyl ether (1:2, v/v) by refluxing on a water bath for 30
min, placed in an ultrasonic bath for 10 min.

Extraction was repeated two times

The total organic solvent was combined and removed with a rotary
evaporator at 40°C under vacuum and the residue was dissolved in 10
mL methanol

The methanol solution was filtered through a 0.45-µm membrane filter

for GC–MS analysis.
The reason an analytical method was used

• GC-MS is quick and sensitive method

• Use SIM with 3 areas because in Lv et al.
(2000) research, they used m/z 136 in GC-MS,
It which was not an accurate and simple
method, TMP in Ma-Huang has never been
simultaneously determined with ephedrine
alkaloids
A = identification
all standard
with Total ion
chrom (TIC)
PE, E, and small
detected NE
and NPE

B = identification
all standard
with m/z 58
PE and E
C = identification
all standard
with m/z 44
detected all

D = identification
all standard
wit h m/z 136
PE , E, NE, and
NPE (TMP not
showed)
 Validation
linearity %Recovery
y = (1.46 × 107)x – (2.80 × 106) E = 105 %
(r = 0.9985) for E PE = 106 %
y = (1.76 × 107)x – (3.31 × 106) TMP = 93%
(r = 0.9970) for PE
y = (4.41 × 104)x – (2.14 × 104)
(r = 0.9992) for TMP Presision
E = 1.5%
LOD PE = 1.2%
E = 0.4 ng TMP = 2.0 %
PE = 0.7 ng N=4
TMP = 0.02 ng
 Data obtained, are they good data ?
• The final data was good, but the detail data
was not showed, so we can’t analyse the data
 Result and discussion, was the experiment
capable of proving and obtaining the data
corresponding the purpose ?
• The purpose was not capable of proving and
obtaining the data corresponding because not
all alkaloids can detected, the peaks of NE and
NPE were tailed and the contents of NE and
NPE in Ephedra sinica Stapf were very low, NE
and NPE were not quantitated in this study.
 Conclusion
• Ephedrine alkaloids and TMP were
simultaneously determined by GC–MS without
tedious pretreatment, such as derivatization.
• Both the full-scan acquisition mode and SIM
mode were used to identify and quantitate the
alkaloids in Ephedra sinica Stapf.
• It was a simple, reliable, and quick method for
the determination of the active components in
ephedrine-containing medicinal herbs.
Do you agree/disagree that your paper
is a good research paper ? Why ?
• Disagree, because the data in this paper not
showed detail data