Simultaneous determination of 22 marker compounds in traditional herbal medicine Ojeoksan using UPLC-MS/MS systems
- 주제(키워드) Traditional herbal medicine , Ojeok-san , Validation , UPLC-MS/MS , Marker compound , Simultaneous analysis
- 주제(DDC) 651.1
- 발행기관 아주대학교 일반대학원
- 지도교수 백승훈
- 발행년도 2024
- 학위수여년월 2024. 2
- 학위명 석사
- 학과 및 전공 일반대학원 바이오헬스규제과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000033759
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Herbal medicines (HMs) have a long history of medicinal use with safety and efficacy against various diseases and have contributed as major sources of modern medicines. Traditional herbal medicines (THMs), the prescriptions consisted of various HMs, have long been used as cures for diseases. Some of them are still used as tonics or complementary medicines. Multiple components are one of the main advantages of THMs for multi-targeting, but considered as hurdles for the quality control of THMs in the regulatory framework of modern medicine. Ojeoksan (OJS), the second most prescribed herbal medicine in Korea, consisted of 16 herbal substances and are the THM used as pain-relieving agent. Until now, quantitative methods have been performed with only a few marker compounds of the HMs that make up OJS, but no method has been established to set and confirm marker compounds of all constituent HMs. The aim of this study is to establish an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) assay for simultaneous determination of 22 marker compounds in OJS. The analytes were separated in liquid chromatography using an Infinitylab Poroshell 120 EC-C18 (2.1 × 150 mm, 2.7 μm) column and gradient elution of the mobile phases 0.1% formic acid and methanol containing 0.1% formic acid. Analytes were detected in a tandem mass spectrometry using both negative and positive electrospray ionization (ESI) mode and scheduled multiple reaction monitoring (sMRM) acquisition. Method validation study was carried out using the matrices of OJS tablet and soft-extract for three consecutive days to access intra- and inter-day variance. Validations results showed acceptable performance characteristics, including specificity, linearity (R2 ≥ 0.9956), repeatability (RSD ≤ 6.5%), LOD (0.3–3.6 ng/mL), LOQ (1.0–10.9 ng/mL), precision (RSD ≤ 12.1%), and accuracy (recovery 78.4–108.9%). Established method was applied to five commercial OJS products (1 tablet and 4 soft-extract dosage form). The 14 marker compounds indicating the 12 component HMs were determined in products; paeoniflorin, hesperidine, glycyrrhizin, albiflorin, ephedirine, neohesperidin, cinnamic acid, decursin, platycodin D, ferulic acid, 6-gingerol, atractylenolide III, magnolol, and honokiol. Alternative marker compounds for the 3 unidentified HMs should be tested in our future study. Fast and easy assay method was established with sufficient discrimination power. This method as a new assay can contribute to quality control for OJS products.
more목차
1. Introduction 1
1.1. Traditional herbal medicine 1
1.2. The Korean herbal pharmacopoeia 2
1.3. Ojeoksan 4
1.4. Marker compound of component HMs in OJS 7
1.5. Aim of this study 9
2. Materials and methods 10
2.1. Chemicals and reagents 10
2.2. Ojeoksan samples 10
2.3. Instrumentation 11
2.4. Analytical method development 11
2.4.1. Preparation of standard solution 11
2.4.2. Preparation of samples 12
2.4.3. Liquid chromatography 12
2.4.4. Mass spectrometry 13
2.5. Method validation 13
2.5.1. Specificity 13
2.5.2. Linearity, repeatability, LOD and LOQ 14
2.5.3. Accuracy and precision 14
3. Results and Discussion 15
3.1. UPLC-MS/MS condition 15
3.1.1. Optimization of liquid chromatography condition 15
3.1.2. Optimization of mass spectrometry condition 15
3.1.3. Identification of each marker compound 26
3.2. Method validation 34
3.2.1. Specificity 34
3.2.2. Linearity, repeatability, LOD and LOQ 38
3.2.3. Accuracy and precision 40
3.3. Application to real samples 45
4. Conclusion 48
Acknowledgments 48
Reference 49
국문 초록 55
