A rapid micellar HPLC method for simultaneous ...

第 42 卷 第 3 期 2016 年 6 月

兰 州大学学报（医 学 版） Journal of Lanzhou University（Medical Sciences）

Vol.42 No.3 Jun. 2016

文章编号：1000-2812 (2016) 03-0012-10

A rapid micellar HPLC method for simultaneous separation of olanzapine and its related compound A in tablets Lu Ning-wei1, 2, Li Ning1, An Qiong1, 3, Dong Yu-ming1 1. Institute of Pharmaceutical Analysis, School of Pharmacy, Lanzhou University, Lanzhou 730000, China 2. School Hospital, Lanzhou University of Technology, Lanzhou 730070, China 3. Teaching and Research Section of Biochemistry, Department of Basic Medical Sciences, Hexi University, Zhangye 734000, Gansu, China

Abstract：Objective To establish a rapid method for separation of olanzapine and its related compound A in tablets by micellar high performance liquid chromatography and to discuss the possible separation mechanism. Methods The optimized separation conditions were performed using a mobile phase consisted of water (containing 41.7 mmol/L sodium dodecyl sulfate) acetonitrile (35:65, V/V) at pH 2.50 (adjusted with phosphoric acid), running at 1.20 mL/min through a C8 column at 40 °C. The chromatographic method was validated from specificity, precision, stability, linearity, accuracy, minimum limits of detection and quantification. Results The proposed method was highly specific (resolutions were＞2.03 and the peak purity factor were＞996). The contents of olanzapine in tablets were determined and the limits test of olanzapine related compound A in tablets were examined by the proposed method in 5 minutes. The percentage contents of the labeled amount of olanzapine ranged from 97.1 % to 100.4 % and percentages of the olanzapine related compound A ranged from 0.3 % to 0.6 % in olanzapine tablets. The possible separation mechanism was in detail discussed based on the model of possible interaction among solutes, the mobile phase and stationary phase in the micellar high performance liquid chromatographic system. Conclusion The method was rapid, accurate and sensitive and could be applied to control the quality of olanzapine tablets. Keywords: micellar high performance liquid chromatography; olanzapine; olanzapine related compound A CLC number: R378.11 document code: A doi: 10.13885/j.issn.1000-2812.2016.03.003

胶束液相色谱法同时快速分离片剂中奥氮平及其相关物质 A 路宁维 1，2，李

宁 1，安

琼 1，董钰明 1

1. 兰州大学 药学院 药物分析研究所，甘肃 兰州 730000 2. 兰州理工大学 校医院，甘肃 兰州 730070 3. 河西学院 基础医学部 生化教研室，甘肃 张掖 734000 摘要：目的 目的 建立快速分离奥氮平及其相关物质 A 的胶束高效液相色谱新方法，探讨其分离机理。方法 方法 采用 MicrosorbMV 100 C8 色谱柱，流动相为含 41.7 mmol/L 的十二烷基硫酸钠、pH 2.5 （用磷酸调节） 的胶束溶液—乙腈 （35∶65，V/V）， Received date: 2015-06-08 Foundation item: Supported by the Fundamental Research Funds for the Central Universities of China(lzujbky-2011-93) Biography: Lu Ning-wei(1987-), female, born in Guyuan, Ningxia, Master, researched on medicine analysis, e-mail: [email protected]; Dong Yu-ming(1971-), male, born in Tianshui, Gansu, Professor, PH D, researched on pharmaceutical analysis, e-mail: [email protected], corresponding author

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Lu Ning-wei, et al:

A rapid micellar HPLC method for simultaneous separation of olanzapine and its related compound A in tablets

13

流速 1.2 mL/min，柱温为 40 °C。并对方法进行方法学考察，包括专属性、精密度、稳定性、线性、准确度、最低检测 限、最低定量限。结果 结果 该方法专属性强 （分离度>2.03，峰纯度>996），5 min 内就可以测定不同片剂中奥氮平含量及其 相关物质 A。奥氮平标示量的百分含量为 97.1%~100.4%；奥氮平相关物质 A 的含量为 0.3%~0.6%。建立了溶质、流动 相和固定相三者之间相互作用的胶束高效液相色谱模型，并基于此模型对可能的分离机理进行了详细讨论。 结论 该方 法快速、准确、灵敏，可用于奥氮平片的质量控制。 关键词：胶束高效液相色谱；奥氮平；奥氮平相关物质 A 中图分类号： R378.11

文献标识码：A

Introduction

alytical methods have been built based on the use of

Olanzapine (OLZ) is a new generation of typi-

high performance liquid chromatography (HPLC)

cal antipsychotic drug on mental illness. Its chemical

with ultraviolet (UV) detection[3, 8- 9], electrochemical

name is 2- methyl- 4- (4- methyl- 1- piperazinyl)- 10H-

detection[10-11], and liquid chromatography/tandem

thieno benzodiazinon (Fig.1). Compared with the

mass spectrometry (LC-MS/MS) [12] for the analysis of

old generation of neuroleptics drug, OLZ appears to

OLZ in biological fluids. HPLC with UV detection

have greater affinity for serotonin (5- HT) than for

has been used for the assay of OLZ tablets[13- 14]. Al-

dopamine D receptors and OLZ has a sufficient clini-

though these methods had high sensitivity, ORCA

cal response without extrapyramidal side- effects[1].

was not analyzed and in order to obtain good symme-

OLZ is more useful than the conventional drugs for

try of peaks, tailing- suppressing reagents, such as

treating the negative symptoms of schizophrenia[2- 3]

teramethylenediamin, triethylamine, ammonium and

and has been widely used for the treatment of schizo-

tetramethylammonium, was used in the mobile

phrenia and manic episodes of bipolar disorder [4].

phase to reduce the interaction between the alkaline

1

The chemical name of olanzapine related com-

groups of analytes and silanol groups of stationary

pound A (ORCA) is 5- methyl - 2- ((2- nitrophenyl)

phase. However, residue on the chromatographic col-

amino)-3-thiophenecarbonitrile (Fig. 1). ORCA is re-

umn of tailing-suppressing reagents might cause irre-

garded as an impurity in the raw drug of OLZ and its

versible change or damage to the column, which

limit quantity is 0.1 % in the United States Pharmaco-

would shorten the service life of the column and af-

poeia (USP) 34[5]. It is also one of the principal impu-

fect the separation efficiency[15], and tailing-suppress-

rities in OLZ tablets. Hence, to ensure the efficacy of

ing reagents might also cause contamination to the

therapy, it is important to determine OLZ as well as

pumps and pipes of HPLC system[16]. Rao et al[17] had

control the limit quantity of ORCA in tablets[5].

presented a reversed phase high performance liquid chromatographic (RP- HPLC) method for simultaneous separation and determination of OLZ and its process impurities in bulk drugs and pharmaceutical formulations. But the retention time of OLZ was a little long (13 minutes). Meng et al[15] had developed a stability- indicating RP- HPLC method with UV detec-

Fig.1 Chemical structures OLZ and ORCA

tion for determination of OLZ and its eight related

Compared to the separation of OLZ related impu-

impurities in the raw material of OLZ. It obtained

rities, extensive work has been done on the analysis of

good resolution for OLZ and the eight impurities.

OLZ. Some early developed methods used gas chro-

However, the retention time of ORCA was nearly 40

[6- 7]

matography (GC) method

for the analysis of OLZ

in a postmortem. While in the last few years, most an-

minutes. The tailing factor of OLZ was 1.42 even when triethylamin was added in the mobile phase.

兰 州大学学报（医 学 版）

14

Micellar liquid chromatography (MLC) is able to solve some of these problems, such as residue of tailing- suppressing reagents, long retention time, asymmetry of the peaks, due to its particular mobile phase. The mobile phase of MLC is composed of a surfactant at a higher concentration than the critical micellar concentration (CMC) [18]. This mobile phase has several advantages compared with conventional hydro- organic eluents such as low cost, low volatility and enhanced separation selectivity using organic solvents[19]. It can also get good symmetry without tailing suppressing regent. Moreover, the main strength of MLC lies precisely in the capability of performing and controlling the separation of mixtures of cationic, anionic, and uncharged polar and non- polar solutes, using isocratic elution[20]. Sodium dodecyl sulfate (SDS) is a widely used anionic surfactant in micellar media given its

2

第 42 卷

Experimental

2.1 Apparatus Dionex UltiMate 3000 HPLC with ultimate 3000 DAD and column compartment is controlled by the chromatographic workstation Chromeleon Client (Dionex Corporation, Sunnyvale, America); Microsorb-100 MV C8 column (250 mm × 4.6 mm, 5 μm) (Varian Company, Palo Alto, America) was employed for the separation of samples; KH- 300 DB numerical controlled ultrasonic cleaning device (Kun Shan He Chuang Ultrasonic Instruments Co LTD, Kun Shan, China) was used to dissolve samples; PHS-3 C pH meter (Shanghai Leici Instrument Plant, Shanghai, China) was used to measure pH of mobile phase; TGL-16 C centrifuge (Shanghai AnTing Scientific Instruments Plant, Shanghai, China) was used for processing samples.

good solubility in water, low CMC and ability that being easily removed from the chromatographic sys-

2.2 Chemical and reagents

tem[21]. Several analytical methods based on micellar

OLZ (No. 1478301) and ORCA (No. 1478321)

media using SDS and an organic modifier have been

reference standards were purchased from United

developed to determine drugs in biological fluids,

States Pharmacopeia Committee. ZYPrexa tablets

such as tamoxifen in plasma[22] and verapamil in

(5 mg/tablet), XiMin tablets (5 mg/tablet) and Ou-

serum[23]. In addition, polyoxyethylene 23 lauryl

LanNing (10 mg/tablet) tablets were purchased from

ether (Brij- 35), a nonionic surfactant, has been also

Eli Lilly Company, Indianapolis, America, Chang-

applied as micellar media in MLC [24].

zhou Watson Pharmaceutical Co LTD, Changzhou,

Though MLC has been used to analyze kinds of

China and Stockhausen Pharmaceutical Co LTD, Li-

samples, to the best of our knowledge, none of meth-

anyungang, China, respectively. Acetonitrile and

ods were reported by micellar high performance liquid chromatography (MHPLC) with diode array ultraviolet detector (DAD) to determine OLZ and limit test of ORCA in tablets. The retention mechanisms of MLC were discussed by Armstrong et al[25- 28], but the mechanism of OLZ and ORCA in the proposed method has some difference with that of them. In this work, we developed a simple, accurate and rapid MHPLC method with DAD for the quanti-

methanol (HPLC grade) were purchased from Shandong YuWang Industrial Co LTD (Jinan, China). Phosphoric acid (analytical grade) was purchased from Tianjin FuYu Chemical Co LTD (Tianjin, China). Distilled water was obtained from the GLP Lab of Lanzhou University (Lanzhou, China). SDS (analytical grade) was purchased from Shanghai ZhongQin Chemical Co LTD (Shanghai, China). 2.3 Standard solutions preparation

tative determination of OLZ and limit test of ORCA

OLZ (1 000 μg/mL) and ORCA (1 000 μg/mL)

in tablets for the first time. In addition, the separa-

reference standard solutions were prepared respec-

tion mechanism of OLZ and ORCA of the proposed

tively in acetonitrile as standard stock solutions. All

method was discussed.

standard stock solutions were stored until use under

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Lu Ning-wei, et al:

A rapid micellar HPLC method for simultaneous separation of olanzapine and its related compound A in tablets

refrigeration at 4 °C. The stock solutions of OLZ and ORCA refer-

15

vestigated to gain better separation and a stable baseline in chromatograms within a short analysis time.

ence standard were quantitatively transferred with

As shown in Fig. 2A, the retention time of OLZ

the mobile phase consisted of water (containing

and ORCA was delayed especially that of OLZ when

41.7 mmol/LSDS)-acetonitrile (35∶65, V/V) at pH 2.5

the concentration of SDS was increased. The resolu-

(adjusted with phosphate) to get standard solution of

tion of solutes was also increased with the increase

50 μg/mL of OLZ and ORCA, respectively.

of concentration of SDS.

2.4 Sample preparation For the OLZ tablets (ZYPrexa, 5 mg/table; XiMin 5 mg/table; OuLanNing 10 mg/table), ten units were weighed accurately, placed into a 25 mL volumetric flaskand each sample was dissolved and diluted to volume with the mobile phase. Then, ultrasonic treatment for 10 min (100 kHz, 40 W), centrifugal separation (3 000 rpm), and supernatant was as the stock solution. For the tablets, final concentrations of the stock solutions, were 1 000 μg/mL. Aliquots of the stock solutions were taken and diluted with the mobile phase to get solutions of 100 μg/mL and 10 μg/mL, respectively. 2.5 Optimized chromatographic conditions

As shown in Fig. 2B, the retention time of the two peaks reduced and the sensitivity of them increased when the proportion of acetonitrile increased from 50 % to 70 %. The retention time of OLZ was reduced significantly greater than that of ORCA, which led to the resolution of them decrease. In addition, when acetonitrile proportion was 20%, the peak of OLZ would be bifurcated (as shown in Fig.3); when the ratio was 75 %, the resolution of OLZ and ORCA became poor (as shown in Fig. 4). Separation could not be obtained. As shown in Fig. 2C, with the increasing of mobile phase pH, the retention time of OLZ was delayed and the peak of OLZ became short and wide, and the peak of ORCA had almost no change. To

Quantification of OLZ and limit test of ORCA

obtain short retention time, satisfactory resolution of

were performed using a mobile phase consisted of

OLZ and ORCA and high column efficiency,

water (containing 41.7 mmol/L SDS) - acetonitrile

41.7 mmol/L SDS, 65% acetonitrile and pH 2.5 were

(35∶65, V/V) at pH 2.5 (adjusted with phosphoric ac-

selected for further studies.

id) running at a constant flow of 1.2 mL/min. The

MHPLC usually has low column efficiency[29].

mobile phase was filtered through 0.22 μm Millipore

Two ways could improve the column efficiency. One

filtration. The column temperature was kept constant

way is to raise the column temperature and the other

at 40 ± 0.5 ° C. The detection wavelength was set at

is to increase the proportion of organic solvent in mo-

260 nm. The injection volume was 10 μL.

bile phase. But when the column temperature was

3

Results

3.1 Optimization of chromatographic conditions A single rotation method was used to optimize

too high, the life of the analytical column would be decline

[26]

. For obtaining high column efficiency and

weak damage to column, 40 ° C was selected as the optimized temperature in this method.

the chromatographic conditions and the factors were

The results indicated that the retention time of

investigated one by one. The concentration of SDS

OLZ and ORCA was delayed when the flow rate

(4.11, 8.23, 2.00, 2.46, 4.17, 4.91 mmol/L), the pro-

was decreased. The peak height and column pressure

portion of acetonitrile (20%, 50%, 55%, 60%, 65%,

became somewhat constant with the increasing flow

70% , 75% ), the pH of mobile phase (2.0, 2.5, 3.0,

rate. In order to obtain higher peak height, lower col-

6.0), flow rate (0.8, 1.0, 1.2, 1.5, 1.8, 2.0 mL/min),

umn pressure and rapid analytical time, the flow rate

and column temperature (35, 40, 45, 50 °C) were in-

was finally set as 1.2 mL/min.

16

兰 州大学学报（医 学 版）

第 42 卷

Fig.3 The chromatograms of OLZ and ORCA. Chromatographic conditions: the ratio of organic solvents was 20%. The other conditions are same as the Fig. 2B.

Fig.4 The chromatograms of OLZ (b) and ORCA (a). Chromatographic conditions: the ratio of organic solvents was 75%. The other conditions are same as the Fig. 2B.

3.2 Validation of the method The proposed method was validated with respect to specificity, precision, stability, linearity, limit of detection (LOD) and limit of quantification (LOQ). Under the optimized chromatographic conditions, the tailing factor for OLZ and ORCA were 1.29 and 1.32, the resolution for OLZ and ORCA were 2.5 and 3.1, and the peak purity match factor The mixture mixed standard solution contained 50 μg/mL OLZ and 50 μg/mL ORCA. Chromatographic conditions: the flow rate was 1.2 mL/min and column temperature was 40 °C. In Fig. 2A, the ratio of organic solvents was 65% and the pH was 2.5. In Fig. 2B, the concentration of SDS was 41.7 mmol/L and the ratio of organic solvents was 65%. In Fig. 2C, the concentration of SDS was 41.7 mmol/L and the pH was 2.5.

Fig.2

The chromatograms of OLZ (b) and ORCA (a) under the different concentration of SDS (A), different organic solvents (B), and different pH (C).

for OLZ and ORCA were 996 and 999. The two analytes were separated in 5 min. 3.2.1 Specificity The method specificity was determined employing a diode- array detector under the optimized chromatographic conditions. The two analytes were well separated, resolutions were > 2.03 between adja-

Lu Ning-wei, et al:

第3期

A rapid micellar HPLC method for simultaneous separation of olanzapine and its related compound A in tablets

17

cent peaks, and the peak purity factor were > 996.

0.8 % , respectively. It indicated that the OLZ and

Based on these results, the specificity of the pro-

ORCA in the mobile phase was stable for at least 32 h.

posed HLIC method was confirmed.

3.2.4 Linearity and range

3.2.2

The calibration curves for OLZ and ORCA

Intra-day and inter-day precision The method precision was established by inject-

were established by using the areas of chromato-

ing six standard samples of OLZ and ORCA at

graphic peaks obtained from five different concentra-

100 % level of the expected concentrations of OLZ

tions (18.8, 37.6, 75.2, 150, 301 μg/mL for OLZ;

and OR CA in the tablet samples for the intra- day

2.27, 5.54, 11.1, 22.1 , 44.3 μg/mL for ORCA). The

precision and across three different days for the

linear relationships between the concentrations OLZ

inter- day precision. The relative standard deviations

and ORCA and the corresponding peak areas were

(RSDs) of peak area and retention time for OLZ and

summarized in Table 1 and demonstrated good lin-

ORCA were below 1.3 % and 0.2 % , respectively.

earity for OLZ (r2=0.999 0) and ORCA (r2=0.999 1).

As shown in Table 1, it suggested the method had

3.2.5 LOD and LOQ

good reproducibility. 3.2.3

The LOD and LOQ for OLZ and ORCA were

Stability

estimated at a signal-to-noise ratio of 3∶1 and 10∶1,

The stability of OLZ and ORCA in the mobile

respectively, by injecting a series of diluted solutions

phase was determined during 0, 2, 4, 8, 16, 32 h

with known concentration. As shown in Table 1, the

under the optimized chromatography conditions. The

results demonstrated good sensitivity for the analysis

RSDs of OLZ and ORCA peak area were 1.8 % and

of OLZ and ORCA.

Table 1 Range/ (μg/mL) OLZ

LOD/ LOQ/ (μg/mL) (μg/mL)

Linearity

A=0.8690 C+0.4439 (r2=0.999 9) A=0.1601 C－0.0096 2.27～44.3 2 (r =0.999 1) 18.8～301

ORCA

Results of method validation Intra-day precision

Inter-day precision

RSD % RSD % (Retention time) (Peak area)

RSD % RSD % (Retention time) (Peak area)

0.03

0.15

0.1

1.2

0.2

1.3

0.80

2.40

0.2

0.8

0.2

0.8

RSD: relative standard deviation.

3.2.6 Accuracy

where m1 and m2 are the measured quantity before

The accuracy of the method was determined

adding standard and after adding standard, respec-

with recovery studies by using the standard addition

tively, m is the quantity of adding standard. The ob-

method. Recovery of OLZ was evaluated at three dif-

tained recoveries for OLZ ranged from 93.7% to

ferent concentrations, 80, 100, 120 μg/mL in tablets,

116.8% (Table 2).

respectively. Each level was repeated three times, and the percentage recoveries were calculated ac-

3.3 Application The validated method was applied to quantify

cording to the follow equation: Recovery (%) = (m2－m1)/m × 100 Table 2

ZYPrexa

(1)

OLZ and limit test of ORCA in tablets (XiMin

Recoveries of olanzapine in three tablets

Low concentration (80 μg/mL) Repeatability Recovery/% (RSD %) 91.8 0.6

Middle concentration (100 μg/mL) Repeatability Recovery/% (RSD %) 116.8 1.9

High concentration (120 μg/mL) Repeatability Recovery/% (RSD %) 116.6 2.8

XiMin

93.7

2.7

104.4

1.9

113.4

7.3

OuLanNing

91.4

0.9

101.9

5.5

112.8

2.7

兰 州大学学报（医 学 版）

18

第 42 卷

(Fig. 5A), ZYPrexa (Fig. 5B) and OuLanNing (Fig.

high concentration sample and the peak area of OLZ

5C)). The peak area of OLZ and ORCA were ob-

in low concentration sample. As shown in Fig. 5 the

tained from the MHPLC chromatogram. The sample

excipients in the tablets had no interference to the sepa-

solutions were prepared at 100 μg/mL for determination of OLZ in different tablets. Then, its concentration was calculated by external standard method based on the established calibration curve in table I.

ration of OLZ and ORCA and ORCA could be detected successfully. The OLZ percentage contents of the labeled amount ranged from 97.1% to 100.4% and

Contrast of high (1 000 μg/mL) and low (10 μg/mL)

ORCA percentages ranged from 0.3 % to 0.6 % for

concentration of samples was carried out for limit

OLZ tablets produced by different manufactories.

test of ORCA in tablets. Its percentage was ex-

The results indicated that the method could be applied

pressed by the ratio of the peak area of ORCA in

to determine OLZ and limit test ORCA in tablets.

Chromatographic conditions: mobile phase consisted of water (containing 41.7 mmol/L SDS) -acetonitrile (35∶65, V/V) at pH 2.50. The flow rate was 1.20 mL/min. The column temperature was kept constant at 40±0.5 °C. The detection wavelength was set at 260 nm.

Fig.5 Chromatograms of OLZ tablets

4

Discussions

4.1 Concentration of SDS on the retention behavior of OLZ Solute reservation in the MHPLC depends on the balance among with water phase, micellar phase

lead the interaction between OLZ and the modified stationary phase strengthened, as shown in Fig. 2A, the retention time of OLZ was delayed. 4.2 Organic solvents on the retention behavior of OLZ

and the modified stationary phase[25]. In this study,

Adding organic solvents into mobile phase

SDS, an anionic surfactant, was employed to form

would change the balance among solutes, stationary

micelle in the mobile phase. The interaction of OLZ

phase, micellar phase and the aqueous phase. The re-

and micelle phase, OLZ and modified stationary

tention of solutes depends on the balance between

[25, 30] . phase made comprehensive decision to the solutes’ hydrophobic interaction and electrostatic force When the proportion of acetonitrile increased in chromatographic behavior. OLZ is an alkaline com-

pound which formed cations in acid conditions. The

the mobile phase, surfactant adsorption on the sta-

outlet layer of SDS micellar group has negative

tionary phase might be greatly decreased. Negative

charge. There might be electrostatic attraction be-

charge on stationary phase would be reduced. Elec-

tween the positive charge of the OLZ and the nega-

trostatic attraction between cations of OLZ and sta-

tive charge of micelle surface. When the concentra-

tionary phase decreased. OLZ distribution in the sta-

tion of SDS was increased, the adsorption of SDS on

tionary phase decreased and the retention time re-

stationary phase might be increased which would

duced (Fig. 2B).

第3期

Lu Ning-wei, et al:

A rapid micellar HPLC method for simultaneous separation of olanzapine and its related compound A in tablets

4.3 Mobile phase pH on the retention behavior

19

From the structure of ORCA, the cyan group

of OLZ

made the molecule hydrophilic. And the interaction

OLZ is an alkaline compound and exists an inher-

between ORCA and water- acetonitrile phase was

ent ionization equilibrium. When the acidity changed,

also hydrophobic interaction, except electrostatic at-

the ionization equilibrium was also changed and thus

traction. From the structure of OLZ and ORCA, the

affected the distribution of OLZ in micellar phase,

hydrophilia of ORCA was stronger than OLZ, so

stationary phase and water phase. When the pH of

the mobile phase has the stronger elutive power for

mobile phase increased, molecular forms of OLZ in-

ORCA than OLZ. This accorded with the experiment

creased and ionic forms of OLZ decreased. The hy-

results that the retention time of ORCA was shorter

drophobic interaction between OLZ and stationary

than OLZ.

phase increased and the electrostatic attraction with micelle surface weakened, which made retention time of OLZ delayed (Fig. 2C). In addition, when the pH of mobile phase was increased, the sensitivity and column efficiency were significantly reduced for OLZ. 4.4 Changes of ORCA retention behavior From the chemical structure of ORCA (Fig. 1), Fig.6 Model of possible interaction among solutes, the mobile phase and stationary phase in MHPLC system of this study.

we can conclude that ORCA has large polarity because it has cyano and nitro groups. Because water and acetonitrile are polar solvents, ORCA might in-

5

Conclusion

teract strongly with water and acetonitrile in the mobile phase by hydrophobic interaction. ORCA

A simple MHPLC method was established and

flowed out with water- acetonitrile phase and the

validated for the determination of OLZ and limit test

changes of the SDS concentration, acetonitrile pro-

of ORCA in tablets. The established method was

portion and mobile phase pH had little influence on

found to be rapid (in 5 minutes), sensitive, precise,

ORCA retention.

and accurate. The possible separation mechanism

4.5 Model of separation mechanism Based on the above results, we simulated a simple model of micellar chromatography environment for the separation of OLZ and ORCA (Fig. 6). There are stationary phase, micellar phase and water-acetonitrile phase in the chromatographic system. The surface of the stationary phase was modified by SDS monomers[31]. Because the hydrophilic end of SDS monomers has negative electricity, the surface of modified stationary phase has also negative electricity[32].

was discussed. The proposed method was applied to determine OLZ and limit test ORCA in OLZ tablets successfully. The method could be used for the quality control of OLZ tablets. References [1] Navari R M. Olanzapine for the prevention and treatment of chronic nausea and chemotherapy- induced naused and vomiting[J]. European Journal of Pharmacology, 2014, 722: 180-186. [2] Raggi M A, Casamenti G, Mandrioli R, et al. Quantitation of olanzapine in tablets by HPLC, CZE, derivative spec-

The OLZ and ORCA were alkali compound, and

trometry and linear voltammetry[J]. Journal of Pharma-

formed cation in acid conditions. So the interactions

ceutical and Biomedical Analysis, 2000, 23(6): 973-981.

among OLZ, modified stationary phase and micelle phase were electrostatic attraction.

[3] Olesen O V, Linnett K. Determination of olanzapine in serum by high- performance liquid chromatography using

兰 州大学学报（医 学 版）

20

ultraviolet detection considering the easy oxidability of

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