The Reduction of Crystal Formation Time of Vancomycin Using Silica Gel

실리카겔을 이용한 반코마이신 결정화 시간 단축

김성재, 김진현*

공주대학교화학공학부

Received: April 29, 2014 / Revised: July 9, 2014 / Accepted: July 9, 2014

서 론

반코마이신은그람양성세균의세포벽합성을억제하여세 균을괴사시키는당펩타이드계최초의항생물질로

¹⁹⁵⁶

Eli Lilly

Amycolatopsis orientalis (Streptomyces orientalis, Nocardia orientalis)

^{[4, 16].}

에 내성을 갖는 황색 포도상 구균

(Methicillin Resistance

Staphylococcus aureus, MRSA)

^,

널리사용되고있다

^.

^MRSA

서인공삽입물을이용하여시술하는심장수술과정형외과 수술

^,

(ventriculoperitoneal shunt)

[7, 14].

위해서는여러단계의분리및정제공정을거치게된다

^.

이신의함량

^,

엄격하게규정하고있다

^.

USP (United States

Pharmacopeia)

^HPLC

마이신의함량이

^88%

^,

한물질중에서함량이

^4%

^[15].

EP (European Pharmacopoeia)

^93%

^{, USP}

^4%

^.

^,

^.

[5, 8]

^,

니라결정성물질의물성과형상을조절하는분야의핵심기 술에해당된다

^.

(homogeneous nucleation)

⁽

^,

^,

⁾

^.

^.

The Reduction of Crystal Formation Time of Vancomycin Using Silica Gel

Sung-Jae Kim and Jin-Hyun Kim*

Department of Chemical Engineering, Kongju National University, Cheonan 331-717, Republic of Korea

We investigated the effects of silica gels on the reduction of the crystallization time for the purification of vancomycin. The shortest crystallization time for vancomycin was obtained when silica gel with a pore diameter of 40-60 Å and with a particle diameter of 230-400 mesh was used as the material. The use of silica gel as a surface area increasing material dramatically reduced the crystallization time four fold (6 h) when compared with the results where the surface area had not been similarly increased. In addition, the crystal size of vancomycin was decreased with the addition of silica gel. This improved crystallization process has a significant effect on the convenience and feasibility of the purification step for vancomycin.

Keywords: Vancomycin, crystallization, silica gel

*Corresponding author

Tel: +82-41-521-9361, Fax: +82-41-554-2640 E-mail: jinhyun@kongju.ac.kr

© 2014, The Korean Society for Microbiology and Biotechnology

핵생성의경우상대적으로낮은과포화도로인하여핵생 성속도가매우느리다

^.

^,

^[10].

^,

^.

^.

^,

용분야가넓다

^[6].

수들

⁽

^,

^/

^,

^{, pH,}

^,

^,

⁾

^,

산할수있는방법이

²⁰¹⁰

^[14].

마이신결정생성에많은시간

^(~24

⁾

^.

²⁰¹¹

²⁰¹²

(surface area per volume of reaction solution)

간을

^~12

^{[11, 12].}

는반응액부피당표면적이증가된반코마이신결정화공정 에서새로운표면적증가물질인실리카겔을이용하여반코 마이신결정화시간을획기적으로단축시킬수있는가능성 을조사하였다

^.

^.

재료 및 방법

반코마이신 시료

본실험에사용된반코마이신은토양으로부터분리한미

생물인

N. orientalis

^.

균체를제거하고

^,

하였다

^[13].

양이온교환수지를연속통과하여암모니아로용출한뒤

^,

⁽

^: 88%)

^[14].

결정화 방법

결정화공정은결정화장치내부의여러표면

⁽

^,

⁾

^,

^,

다

^.

^{Fig. 1}

내었다

^.

⁽

^{: 88%)}

^{1 N}

pH 2.5,

^{20 ms/cm}

증류수에용해하였다

^.

^pH

^pH

^.

^[14]

교반속도

^{(640 rpm)}

어뜨렸다

^.

¹⁰

^C

^.

⁽

^,

^,

^,

⁾

⁸

(Sigma-Aldrich, USA)

^{Table 1}

^.

(0.1185 cm

/g)

^{(60.75 m}

^/g)

^.

^{6, 9}

^,

(150 mm, Whatman,

Buckinghamshire, UK)

하여결정표면에불순물을제거하였다

^.

반코마이신 결정 형태 및 크기 분석

반코마이신결정형태및크기측정을위하여전자현미경

(SV-35 Video Microscope System, Some Tech., Korea)

^.

⁽

⁵⁰⁰⁾

^.

^IT- Plus system (Some Tech., Korea)

^.

SEM 분석

Scanning Electron Microscope (MIRA LMH, Tescan,

Czech)

얻어진 반코마이신 결정 형태와 입자표면을 관찰하였다

^. Acceleration voltages

^{20 kV}

^{1 mg}

^.

Fig. 1. Schematic diagram of increased surface area crystal-

lization using silica gel for purification of vancomycin. The

crystallizer size and experimental volume are 90 ml and 13.5 ml,

respectively.

결과 및 고찰

반코마이신결정화에서표면적증가물질로이온교환수지를

사용하면표면적증가가없는경우

^(control)

성에소요되는시간을

²

⁽¹²

⁾

^.

로제공하는요소로작용하기때문이다

^[12].

새로운표면적증가물질로실리카겔을이용하여반코마이신 의결정화에소요되는시간을획기적으로단축시키고자하 였다

^.

⁽

^,

^,

^,

⁾

하기위하여

^,

⁸

^(Table

1)

. 0.1 g/4.5 ml

^.

동일한표면적

^{(60.75 m}

⁾

성된결정을전자현미경으로관찰한결과를

^{Fig. 2A}

^.

silica gel 4

^silica gel 6

⁶

, silica gel 3

⁹

서는결정이생성되지않았다

. Silica gel 1-5

이신결정생성

^/

. Silica gel 4(

: 230-400 mesh)

^.

(silica gel 1-4)

silica gel 5

^silica

gel 4

⁶

렷한결정이생성되지않았다

^.

파악하기위하여

, silica gel 5

(~15

⁾

^.

¹⁵

이결정입자가지사이에끼어있음을확인할수있었다

^.

^,

^.

(silica gel 4

silica gel 6-8

⁾

silica gel 4

^silica

gel 6

되었다

^.

^,

^{40-60 Å}

^.

^[14]

과비교해보면표면적증가물질

⁽

⁾

^1.5- 2.5

(<150 mm radius)

^.

침전물의크기는더작아지는것이다

[2, 9, 10, 17].

정에서임계자유에너지와임계반경의크기는과포화농도의 함수로표시되어있는데과포화농도가증가할수록임계자 유에너지는낮아지고임계반경은작아진다

^.

생성의

^barrier

이루어질수있음을의미한다

^[10].

동일한기공부피

^{(0.1185 cm}

⁾

후생성된결정모양을

^{Fig. 2B}

^.

일하게하였을경우와마찬가지로

silica gel 4

⁶

⁶

, silica gel 3

⁹

^.

^.

^,

^,

230-400 mesh, 40-60 Å

다

. Silica gel 4

겔의표면을

^SEM

^,

실리카겔의표면이매끄러운상태인반면결정화후에는실

리카겔의표면은매우거칠어져있었다

(data not shown).

정화후에실리카겔표면에는많은입자들이생성

^/

Table 1. Physical properties of silica gels.

Type Particle size mesh

( µm)

Pore diameter (Å)

Pore volume (cm

/g)

Surface area (m

/g)

Silica gel 1 35-60 (203-447) 60 0.76 525

Silica gel 2 60-100 (140-203) 60 0.75 480

Silica gel 3 70-230 (65-185) 60 0.80 550

Silica gel 4 230-400 (35-65) 60 0.79 405

Silica gel 5 >400 (<35) 60 0.60 500

Silica gel 6 230-400 (35-65) 40 0.45 570

Silica gel 7 230-400 (35-65) 100 0.89 352

Silica gel 8 230-400 (35-65) 150 1.15 300

Fig. 2. Video microscope images of vancomycin crystals obtained through the crystallization process at a fixed (A) surface area

(60.75 m

) and (B) pore volume (0.1185 cm

) of silica gel. The box drawn with a solid line indicates the crystal structure of vancomycin

obtained through the crystallization process. Scale bar indicates 100 µm.

음을알수있었다

^.

것으로판단된다

^[1].

가로반코마이신결정생성이촉진되고이로인하여결정화 효율이향상되는것으로사료된다

. Silica gel 4

silica gel 6

^SEM

^.

^SEM

도반코마이신결정이성공적으로생성됨을확인할수있었

다

^{(Fig. 3).}

^,

0.1 g/4.5 ml (1 ml

^{+ 3.5 ml}

⁾

silica gel 4

(0.075, 0.150, 0.225 g)

^.

^{6, 9, 12}

silica gel 4

^{Fig. 4}

. Silica gel

4

0.075 g

9

는반면

silica gel 4

0.150, 0.225 g

⁶

^.

Fig. 3. SEM images of vancomycin crystals. (A) Silica gel 4, (B) Silica gel 6.

Fig. 4. Video microscope images of vancomycin crystals obtained at various amounts of silica gel 4. Scale bar indicates 100 µm.

크기는첨가한

silica gel 4

^.

^{[2, 3].}

요 약

본연구에서는 반응액 부피당 표면적

(surface area per volume of reaction solution)

^.

^,

^{40-60 Å,}

230-400 mesh

^.

^.

⁴

^.

^.

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