[논문]Effect of HPLC Analytical Procedure upon Determining Drug Content in PLGA Microspheres

Heo, Sun-Ju; Lee, Hong-Hwa; Lee, Min-Jung; Sah, Hong-Kee

doi:10.4333/kps.2010.40.3.193

Effect of HPLC Analytical Procedure upon Determining Drug Content in PLGA Microspheres 원문보기

Journal of pharmaceutical investigation, v.40 no.3, 2010년, pp.193 - 200

Heo, Sun-Ju (College of Pharmacy, Ewha Womans University) , Lee, Hong-Hwa (College of Pharmacy, Ewha Womans University) , Lee, Min-Jung (College of Pharmacy, Ewha Womans University) , Sah, Hong-Kee (College of Pharmacy, Ewha Womans University)

Abstract ▼ AI-Helper

The objective of this study was to investigate the effects of sample preparation, HPLC conditions and peak measurement methods upon determining progesterone content of poly-d,l-lactide-co-glycolide microspheres. A series of the microspheres with different formulations was first prepared. To determine their actual drug contents, the microspheres were dissolved in tetrahydrofuran and diluted with various amounts of methanol to precipitate the polymer. After removal of polymeric precipitates, the filtrates were subject to HPLC analysis under versatile experimental conditions. Interestingly, the composition of a sample solution (e.g., the ratio of methanol to tetrahydrofuran) affected the magnitudes of both peak fronting and peak broadening of progesterone. Its peak became broader and more asymmetrical at lower methanol:tetrahydrofuran ratios. Furthermore, its peak height was influenced by the proportion of tetrahydrofuran in a sample solution. Such problems encountered with tetrahydrofuran were exacerbated when a larger volume of the sample solution was injected onto an analytical column. Under our experimental conditions a peak area measurement provided more accurate and reliable determination of progesterone content in various microspheres than a peak height determination. Optimizing the composition of a sample solution, HPLC chromatographic conditions and peak analysis methods was a prerequisite to an accurate determination of progesterone encapsulated within microspheres.

주제어

AI 본문요약
AI-Helper

* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.

문제 정의

Changes in peak properties often lead to poor separation and have undesirable effects on the calculation of peak area and/or height for quantitative analysis. Therefore, this study aimed at scrutinizing the influence of the composition of a sample solution, the type of a mobile phase and peak analysis methods upon determining the drug content of PLGA microspheres. Progesterone was used as a drug to be encapsulated into the microspheres throughout this study
Interestingly, peak area remained the same despite tetrahydrofuran-triggered changes in the peak shape of progesterone. This was a reason why reasonable accuracy and precision data were obtained with peak area measurements at various methanol:tetrahydrofuran ratios tested in this study.

가설 설정

So far, little attention has been paid to the effects of sample preparation, HPLC chromatographic conditions and peak analysis methods upon the assay accuracy of drug microencapsulation efficiency. In this study it is supposed that the difference between an injection sample solution and a mobile phase might affect the peak shape of progesterone and subsequently its microencapsulation efficiency. Changes in peak properties often lead to poor separation and have undesirable effects on the calculation of peak area and/or height for quantitative analysis.

제안 방법

Our study highlights that in case when their matrices are different from one another, such differences should be carefully evaluated in order to assure the accuracy and precision of an assay. Based on the discussions of the HPLC chromatographic characteristics described above, an analytical methodology for the determination of progesterone content of PLGA microspheres was optimized as follows: 4 mL of tetrahydrofuran was used to dissolve progesterone-loaded microspheres, and 1 part of the solution was diluted with 5 parts of methanol to precipitate out the PLGA polymer. After removal of the polymeric precipitates, 20 µL of the resultant solution having a methanol to tetrahydrofuran ratio of 5:1 was injected onto the analytical column.
Chromatographic measurements were performed on a Shimadzu model LC-20A series HPLC equipped with a binary pump (LC-20AD), a UV/VIS detector (SPD-20A), and an autosampler (SIL-20A). The system was controlled by the Shimadzu LC Solution 1.
Scanning electron micrographs of PLGA microspheres were taken for the assessment of their surface and internal morphology. Some microsphere samples were embedded in an epoxy resin and cross-sectioned to expose their internal structure, prior to sputter-coating.

성능/효과

An example is a case that adjacent peaks near the peak of a test compound have a potential of influencing its peak; in this case, a peak height measurement provides smaller error than does a peak area measurement. Our results showed that both peak area and peak height measurements provided acceptable precision and accuracy, only when the methanol:tetrahydrofuran ratio of a sample solution was 6:0. In the presence of tetrahydrofuran, however, the peak height of progesterone decreased to a great extent.

참고문헌 (19)

Beck, L.R., Ramos, R.A., Flowers, C.E., Lopez, G.Z., Lewis, D.H., Cowsar, D.R., 1981. Clinical evaluation of injectable biodegradable contraceptive system. Am. J. Obstet. Gynecol. 140, 799-806.

상세보기
Benoit, J.P., Courteille, F.,Thies, C., 1986. A physicochemical study of the morphology of progesterone-loaded poly (D,Llactide) microspheres. Int. J. Pharm. 29, 95-102.

상세보기
Cherrak, D., Guernet, E., Cardot, P., Herrenknecht C., Czok, M., 1997. Viscous fingering: a systematic study of viscosity effects in methanol-isopropanol systems. Chromatographia 46, 647-654.

상세보기
Forgacs, E., Cserhati, T., Balogh, S., Kaliszan, R., Haber, P., Nasal,A., 2001. Separation of strength and selectivity of mobile phase by spectral mapping technique. Biomed. Chromatogr. 15, 348-355.

상세보기
Formento, J.L., Moll, J.L., Francoual, M., Krebs, B.P., Milano, G., Renee, N., Khater, R., Frenay, M., Namer, M., 1987. HPLC micromethod for simultaneous measurement of estradiol, progesterone, androgen and glucocorticoid receptor levels. Applications to breast cancer biopsies. Eur. J. Can. Clin. Oncol. 23, 1307-1314.

상세보기
Gaborieau, M., Nicolas, J., Save, M., Charleux, B., Vairon, J.P., Gilbert, R.G., Castignolles, P., 2008. Separation of complex branched polymers by size-exclusion chromatography probed with multiple detection. J. Chromatogr. A. 1190, 215-223.

상세보기
Ha, H., Lee, Y.S., Lee, J.H., Choi, H., Kim, C., 2006. High performance liquid chromatographic analysis of isoflavones in medicinal herbs. Arch. Pharm. Res. 29, 96-101.

원문보기 상세보기
Issaq, H.J., Young, R.M., 1977. Peak area vs peak height in flameless atomic absorption measurements. Appl. Spectrosc. 31, 171-172.

상세보기
Keunchkarian, S., Reta, M., Romero L., Castells, C., 2006. Effect of sample solvent on the chromatographic peak shape of analytes eluted under reversed-phase liquid chromatographic conditions. J. Chromatogr. A. 1119, 20-28.

상세보기
Leamen, M.J., McManus N.T., Penlidis, A., 2004. Refractive index increment (dn/dc) using GPC for a-methyl styrene/methyl methacrylate copolymer at 670 nm in tetrahydrofuran. J. Appl. Polym. Sci. 94, 2545-2547.

상세보기
Mehta, R.C., Thanoo B.C., DeLuca, P.P., 1996. Peptide containing microspheres from low molecular weight and hydrophilic poly(d,l-lactide-co-glycolide). J. Control. Release 41, 249-257.

상세보기
Moshgeghi, A.A., Peyman, G.A., 2005. Micro- and nanoparticulates. Adv. Drug Deliver. Rev. 57, 2047-2052.

상세보기
Pawar, R., Ben-Ari, A., Domb, A.J., 2004. Protein and peptide parenteral controlled delivery. Expert Opin. Biol. Th., 4, 1203-1212.

상세보기
Pucci, V., Bugamelli, F., Mandrioli, R., Luppi, B., Raggi, M.A., 2003. Determination of progesterone in commercial formulations and in non-conventional micellar systems. J. Pharmaceut. Biomed. Anal. 30, 1549-1559.

상세보기
Sah, H., 2000. Ethyl formate æ alternative dispersed solvent useful in preparing PLGA microspheres. Int. J. Pharm. 195, 103-113.

상세보기
Sah, H., Lee, B. J., 2006. Development of new microencapsulation techniques useful for the preparation of PLGA microspheres. Macromol. Rapid Comm. 27, 1845-1851.

상세보기
Schaefer, M.J., Singh, J., 2001. Effect of additives on stability of etoposide in PLGA microspheres. Drug Dev. Ind. Pharm. 27, 345-350.

상세보기
Taylor, P.J., Brown, S.R., Cooer, D.P., Salm, P., Morris, M.R., Pillans, P.I., Lynch, S.V., 2005. Evaluation of 3 internal standards for the measurement of cyclosporine by HPLC-mass spectrometry. Clin. Chem. 51, 1890-1893.

상세보기
Wu, Z., Zhang, C., Yang, C., Zhang, X., Wu, E., 2000. Simultaneous quantitative determination of norgestrel and progesterone in human serum by high-performance liquid chromatography-tandem mass spectrometry with atmospheric pressure chemical ionization. Analyst 125, 2201-2205.

상세보기

저자의 다른 논문 :

LOADING...

원문 URL 링크

DOI : 10.4333/KPS.2010.40.3.193
한국학술정보 : 저널
AccessON : 저널

*원문 PDF 파일 및 링크정보가 존재하지 않을 경우 KISTI DDS 시스템에서 제공하는 원문복사서비스를 사용할 수 있습니다.

오픈액세스(OA) 유형

GOLD(Hybrid)

저자가 APC(Article Processing Charge)를 지불한 논문에 한하여 자유로운 이용이 가능한, hybrid 저널에 출판된 논문

내보내기 메뉴

내보내기 구분

파일저장
인쇄
메일전송

구성항목

기본정보
상세정보

관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관

저장형식

Text(ASCII format)
Excel format
RefWorks Direct Export
RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley

메일정보

받는사람 (필수): @
보내는사람 (선택): @
제목
내용: KISTI 검색결과 이메일 서비스

안내

총 건의 자료가 검색되었습니다.

다운받으실 자료의 인덱스를 입력하세요. (1-10,000)

검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다.

데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요)

다운로드 파일은 UTF-8 형태로 저장됩니다.
파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오.

Text(ASCII format)
Excel format

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

AI-Helper ※ AI-Helper는 을 사용합니다.

AI-Helper

안녕하세요, AI-Helper입니다. 좌측 "선택된 텍스트"에서 텍스트를 선택하여 요약, 번역, 용어설명을 실행하세요.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.

연합인증