Method Article

An In Vitro Dissolution Determination of Multi-Index Components in Tibetan Medicine Rhodiola Granules

DOI:

10.3791/64670

⸱

November 4th, 2022

 ,  ,  ,  ,  ,  , 
1School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, 3Tibet Rhodiola Pharmaceutical Holding Co. Ltd

In This Article

Summary

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

Here, we test the dissolution of Rhodiola granules (RG) in vitro, draw dissolution curves of salidroside, gallic acid, and ethyl gallate in ultrapure water, and fit the curves to different mathematical models. This protocol provides information and guidance for in vivo bioequivalence and in vivo-in vitro correlation studies of RG.

Abstract

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

The composition of the Tibetan medicine Rhodiola granules (RG) is complex, and the overall quality of RG is difficult to determine. Therefore, establishing a method to determine the multi-component in vitro dissolution of RG is of great significance for quality control. This study uses the second paddle method of the fourth general rule 0931 from the Chinese Pharmacopoeia (2020 edition), compliant with apparatus 2 of the United States Pharmacopeia (USP). The dissolution apparatus was set to a rotation speed of 100 rpm with ultrapure water as the dissolution medium. A sample volume of 1 mL was collected at each timepoint. Furthermore, the cumulative dissolution of gallic acid, salidroside, and ethyl gallic acid in RG at different time points was determined by high-performance liquid chromatography (HPLC). Finally, the dissolution curves were drawn, and the curves were fitted to the GompertzMod, the Gompertz, the Logistic, and the Weibull equations. The results showed that the cumulative dissolution of gallic acid in RG was over 80% at 1 min, the cumulative dissolution of salidroside and ethyl gallic acid was over 65% at 5 min, and the cumulative dissolution of each index component decreased after 30 min. The curve fitting demonstrated that the GompertzMod equation was the best-fitting model for each index component of RG. In conclusion, the dissolution test method described in this protocol is simple, accurate, and reliable. It can characterize the dissolution behavior of the index components in RG in vitro, which provides a methodological reference for quality control of RG and quality evaluation of other ethnic compounds.

Introduction

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

In China, the prevalence of cardiovascular diseases continues to rise, and the morbidity and mortality rates of cardiovascular diseases rank first among Chinese residents1. Angina pectoris of coronary heart disease is caused by luminal stenosis due to coronary atherosclerosis, which leads to relatively insufficient coronary blood supply and myocardial ischemia and hypoxia2. In recent years, the curative effect of traditional Chinese medicine in the treatment of coronary heart disease has been recognized by many doctors3.

Traditional Chinese medicine plays an important r....

Access restricted. Please log in or start a trial to view this content.

Protocol

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

1. Solution preparation

  1. Prepare the reference substance stock solution: Weigh 10.6 mg of salidroside, 5.24 mg of gallic acid, and 5.21 mg of ethyl gallic acid separately on an electronic analytical balance and add them individually into a 5 mL volumetric flask. Then, add HPLC-grade methanol to dissolve and dilute to 5 mL. Finally, shake well to obtain the reference substance stock solution with mass concentrations of 2.120 mg/mL, 1.048 mg/mL, and 1.042 mg/mL, respectively.
    NOTE: The reference substance stock solution contains 2.120 mg/mL salidroside, 1.048 mg/mL gallic acid, and 1.042 mg/mL ethyl gallate as the stock solution of eac....

Access restricted. Please log in or start a trial to view this content.

Results

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

In this study, the precision, stability, repeatability, and sample recovery of RG were all within the methodological range specified in Chinese Pharmacopoeia (Volume 4, 2020)12, indicating that the method was feasible. After repeated debugging, it was determined that the elution gradient used in this study had good resolution (Figure 1) for the three index components in RG. The three index components in RG had a good linear relationship within a specific concentration.......

Access restricted. Please log in or start a trial to view this content.

Discussion

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

The dissolution test is an ideal in vitro method to simulate the disintegration and dissolution of solid oral preparations in the gastrointestinal tract15. It is an important index for evaluating and controlling the quality of solid oral preparations. Therefore, the dissolution test plays an essential role in the development of solid drug oral preparations16. In particular, with the development of traditional Chinese medicine (TCM) quality control technology, the d.......

Access restricted. Please log in or start a trial to view this content.

Disclosures

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

The authors have nothing to disclose.

Acknowledgements

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

This work was funded by the National Key Research and Development Program of China (2017YFC1703904), the University (Chengdu University of TCM) - enterprise (Tibet Rhodiola Pharmaceutical Holding Co. LTD) cooperation project (1052022040101); the Regional Innovation and Cooperation Project of the Science & Technology Department of Sichuan Province (2020YFQ0032); and the Key R&D and Transformation Program of the Science & Technology Department of Qinghai Province (2020-SF-C33).

....

Access restricted. Please log in or start a trial to view this content.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
Chromatographic columnZORBAX Eclipse  XDB-C184.6 mm x 250 mm, 5 µm
Drug dissolution testerShanghai Huanghai Pharmaceutical Inspection Instrument Co., Ltd.RCZ-6B3
Electronic analytical balanceShanghai Liangping Instruments Co., Ltd.FA1004
Ethyl gallate (HPLC, ≥98%)Chengdu Desite Biotechnology Co., Ltd.DSTDM006301
Function drawing softwareOriginLab Corporation, Northampton, MA, USA2022
Gallic acid (HPLC, ≥98%)Chengdu Desite Biotechnology Co., Ltd.DSTDM000802
High performance liquid chromatographyAgilent Technologies Singapore (International) Pte. Ltd.Agilent 1260 Infinity figure-materials-1
HPLC grade methanolThermo Fisher Scientific (China) Co., Ltd.216565
InjectorChengdu Xinjin Shifeng Medical Apparatus & Instrument Co., Ltd.0.7 (22 G)
Millipore filterTianjin Jinteng Experimental Equipment Co., Ltdφ13 0.22 Nylon66
Rhodiola granulesTibet Nodikang Pharmaceutical Co., Ltd.210501
Salidroside (HPLC, ≥98%)Chengdu Desite Biotechnology Co., Ltd.DST200425-037
Ultra pure water systemicMerck Millipore Ltd.Milli-Q
Ultrasonic cleansing machineNingbo Xinyi Ultrasonic Equipment Co., LtdSB-8200 DTS

References

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,
  1. Smith, S. C., Zheng, Z. J. The impending cardiovascular pandemic in China. Circulation Cardiovascular Quality and Outcomes. 3 (3), 226-227 (2010).
  2. Wang, D., Wang, P., Zhang, R., Xi, X.

Access restricted. Please log in or start a trial to view this content.

Reprints and Permissions

Request permission to reuse the text or figures of this JoVE article

Request Permission

Tags

Rhodiola GranulesIn Vitro DissolutionMulti Index ComponentsDissolution TestHigh Performance Liquid ChromatographyGallic AcidSalidrosideEthyl GallateDissolution CurveQuality Control

Related Articles