Identification and Determination of Nicorandil and

its Degradation
Products by HPLC and GC/MS
Zhong Zhou CHENG1∗, Ze Hui JIA 1, Yan CHEN 1, Li Ying CHEN 2, Hua LI 1
Pendahuluan
Nicorandil [N-(2-hydroxyethyl)-nicotinamide nitrate NIC] is a novel
kind of compound in the treatment of angina pectoris.
NIC can be degraded easily in storage.
The degradation products include :
N-(2-hydroxyethy) nicotinamide (HN), nitrate ion (NI), and nicotinic
acid (NA),
in Scheme 1. Determination of NIC1-5, HN6-7 and NI8 have been
studied, but determination of NA is rare.
In this paper, we report the identification of NA by HPLC and GC/MS,
and separation of NIC and its three degradation products in HPLC.
The method is suitable to control the quality of nicorandil in
production process and clinical investigation.
Skema 1
Experiment
NIC (purity ≥ 99.93 %) is donated by Xi’an Libang
Pharmaceutical Co., Ltd., raw drug NIC: 971204 (Institute of
Pharmaceutical industry of Shaanxi province), 2001010
(Jiangsu Diyi Pharmaceutical Co., Ltd), NIC tablets: 040601,
031201 and 030801 (Shaanxi Zhengkang Pharmaceutical Co.,
Ltd.),
methanol is in HPLC grade and otherreagents are in
analytical grade. The stocked solution of NIC and HN (350
mg/L), NI (NO3 -1– O: 108 mg/L) and NA (NA– OH: 11 mg/L)
and other samples were prepared by dissolving the
corresponding reagents in water and diluting in mobile
phase.
Experiment
The HPLC experiments were performed using a LC-
10ATVP (Shimadzu Japan) system with diode array
detector (DAD). The Shim-Pack VP-ODS C18 column
was in 1 50x 4.6 mm (i.d.) (5 µm), the mobile phase
was methanol and 0.002 mol/L phosphate buffer (pH
7.0) (3:7, v/v), the flow rate 1 mL / min and injection
volume 20 µL.
Experiment
The GC/MS experiments were performed using a HP 6890 GC
with connection to a HP5973 MS series (Agilent USA).
The column was a HP-5 quartz capillary; the column
temperature was raised from 60℃ to 270℃ at the rate of 15℃
/min, the inlet temperature was 280℃, the carrier gas nitrogen at
a follow-rate of 0.6 mL/min.
The ionization source temperature was 230℃ and the scan range
was 29-450 amu.
NIC and 030801 were dissolved in methanol, respectively, and
detected after 24 hours.
The UV experiments were performed according to the reference
5, using an UV-2450 (Shimadzu Japan) spectrophotometer.
Results and Discussion

The pure NIC sample was one peak in HPLC. Several

peaks of degradation products can be obtained when
NIC was degraded. NA was one of degradation
products of NIC, and it was identified by HPLC with
that of the pure NA. Figure 1 and Figure 2 showed the
retention time and UV spectra of NA which were same
with the pure NA.
Results and Discussion

NA is confirmed by GC/MS again. Figure 3 (C) and

(D) showed the GC and MS result of NA in the 03080.
M/z 106 and m/z 137 in Figure 3 (D) indicated the
fragment of M (NA)-OH and the fragment of wethyl
ester of NA formed with methanol (solvent in
experiment) respectively. The facts indicated that NA
was pure NA.
Results and Discussion

In the 030801, there are four peaks corresponding to

NIC, HN, NI and NA in HPLC, but NI does not appear
in GC (see Figure 3 (C) ) due to it is inorganic ion. HN
and NIC is a peak namely “HN+NIC” because nitrate
ester of NIC was lost and NIC was changed to HN in
GC system. Consequently, NIC and HN of 030801
formed one peak in Figure 3 (C), and its MS data is
consistent with m/z 148 fragment of M(NIC)-NO2-OH
in Figure 3 (B).
Results and Discussion
Results and Discussion
Results and Discussion

The developed method has a good linear correlation

(r) between peak area (X) and concentration (Y) in the
corresponding concentration ranges about NIC and
degradation products by HPLC. The regression
equation, the recovery (n=5) and the limit of detection
(LOD S/N ＞ 10) are showed in Table 1.
Results and Discussion
Table 2 and 3 show the contents in raw drug of NIC
and tablets obtained by HPLC and UV respectively.
The results show NIC is unstable and easy to degrade
in storage, and tablets are more rapidly degraded than
raw drug. As shown in Scheme 1, the degradation
process of NIC in storage is a reverse reaction of its
synthesis9.
Results and Discussion
The total amount of NIC, HN, NI and NT obtained by
HPLC is close to the one obtained by UV value. The
results indicate that the HPLC method is a more
suitable method for the quality control of NIC,
because degradation products of NIC can be separated
completely and determined simultaneously in HPLC
and UV can not.
Results and Discussion
Results and Discussion
 Ciprofloxacin

Ciprofloxacin is an antibiotic used to treat a number of bacterial infections.[2]

This includes bone and joint infections, intra abdominal infections, certain type of
infectious diarrhea, respiratory tract infections, skin infections, typhoid fever, and
urinary tract infections, among others.[2] For some infections it is used in addition to
other antibiotics.[2] It can be taken by mouth, in eye drops, or intravenously.
Vankomicin
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Vancomycin is an antibiotic used to treat a number of bacterial infections.[3] It is
recommended intravenously as a treatment for complicated skin infections,
bloodstream infections, endocarditis, bone and joint infections, and meningitis caused
by methicillin-resistant Staphylococcus aureus.[4] Blood levels may be measured to
determine the correct dose.[5] Vancomycin is also recommended by mouth as a treatment
for severe Clostridium difficile colitis.[3] When taken by mouth it is very poorly absorbed.[
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Sefadroksil adalah sebuah antibiotik spektrum luas jenis sefalosporin yang efektif
dipakai untuk infeksi bakteri Gram-positif dan Gram-negatif.
Sefadroksil adalah antibiotik bakterisida.