Development and Validation of a Simple HPLC-UV Method for Analysis of Harmine

Galip Mesut Demir; Afife Büşra Uğur Kaplan; Mine Gülaboğlu; Meltem Çetin

Article Details

Development and Validation of a Simple HPLC-UV Method for Analysis of Harmine

Authors:

Galip Mesut Demir | Afife Büşra Uğur Kaplan | Mine Gülaboğlu | Meltem Çetin

Publisher:

Karyay Karadeniz Yayımcılık

Main Topics:

Tıbbi ve Biyomoleküler Kimya (Diğer)

Journal:

Black Sea Journal of Engineering and Science Vol.8 Issue 4, pp.1096-1102

Document Type:

Article

Publication Date:

7/2025

Document Language:

English

Document ID (Org No.):

2001346265

pISSN:

2619-8991

Keywords:

Abstract:

 In this study, a rapid, simple, and sensitive HPLC-UV method was developed and validated in accordance with ICH Q2 R1 guideline for the quantitative analysis of harmine HRM, a main β-carboline alkaloid found in the Peganum harmala Uzerlik plant. The method was established based on the UV spectral analysis, which indicated maximum absorbance for HRM at 243 nm. A mixture of methanol and ultrapure water containing 1 formic acid 50:50, vv was selected as the mobile phase. In the separation carried out under isocratic conditions, the retention time of HRM was determined as 5.5 min, and the entire analysis was completed in under 8 min. The method demonstrated excellent linearity within the concentration range of 136 µgmL, with a correlation coefficient R² of 0.99998. The accuracy and precision parameters of the method were found to be within acceptable limits, with relative error RE and relative standard deviation RSD both below 2. The limits of detection and quantification LOD and LOQ were 0.269 µgmL and 0.816 µgmL, respectively. Also, it was determined that HRM standard solutions were stable for up to 7 days when stored at different temperatures 20C, 28C, 25C. In conclusion, this method offers a reliable alternative for the quantitative determination of HRM with high analytical quality, reproducibility, and short time. It can be widely used in routine laboratory practices for the monitoring of HRM in pure form and in various pharmaceutical formulations.

 In this study, a rapid, simple, and sensitive HPLC-UV method was developed and validated in accordance with ICH Q2 R1 guideline for the quantitative analysis of harmine HRM, a main β-carboline alkaloid found in the Peganum harmala Uzerlik plant. The method was established based on the UV spectral analysis, which indicated maximum absorbance for HRM at 243 nm. A mixture of methanol and ultrapure water containing 1 formic acid 50:50, vv was selected as the mobile phase. In the separation carried out under isocratic conditions, the retention time of HRM was determined as 5.5 min, and the entire analysis was completed in under 8 min. The method demonstrated excellent linearity within the concentration range of 136 µgmL, with a correlation coefficient R² of 0.99998. The accuracy and precision parameters of the method were found to be within acceptable limits, with relative error RE and relative standard deviation RSD both below 2. The limits of detection and quantification LOD and LOQ were 0.269 µgmL and 0.816 µgmL, respectively. Also, it was determined that HRM standard solutions were stable for up to 7 days when stored at different temperatures 20C, 28C, 25C. In conclusion, this method offers a reliable alternative for the quantitative determination of HRM with high analytical quality, reproducibility, and short time. It can be widely used in routine laboratory practices for the monitoring of HRM in pure form and in various pharmaceutical formulations.

 In this study, a rapid, simple, and sensitive HPLC-UV method was developed and validated in accordance with ICH Q2 R1 guideline for the quantitative analysis of harmine HRM, a main β-carboline alkaloid found in the Peganum harmala Uzerlik plant. The method was established based on the UV spectral analysis, which indicated maximum absorbance for HRM at 243 nm. A mixture of methanol and ultrapure water containing 1 formic acid 50:50, vv was selected as the mobile phase. In the separation carried out under isocratic conditions, the retention time of HRM was determined as 5.5 min, and the entire analysis was completed in under 8 min. The method demonstrated excellent linearity within the concentration range of 136 µgmL, with a correlation coefficient R² of 0.99998. The accuracy and precision parameters of the method were found to be within acceptable limits, with relative error RE and relative standard deviation RSD both below 2. The limits of detection and quantification LOD and LOQ were 0.269 µgmL and 0.816 µgmL, respectively. Also, it was determined that HRM standard solutions were stable for up to 7 days when stored at different temperatures 20C, 28C, 25C. In conclusion, this method offers a reliable alternative for the quantitative determination of HRM with high analytical quality, reproducibility, and short time. It can be widely used in routine laboratory practices for the monitoring of HRM in pure form and in various pharmaceutical formulations.

 In this study, a rapid, simple, and sensitive HPLC-UV method was developed and validated in accordance with ICH Q2 R1 guideline for the quantitative analysis of harmine HRM, a main β-carboline alkaloid found in the Peganum harmala Uzerlik plant. The method was established based on the UV spectral analysis, which indicated maximum absorbance for HRM at 243 nm. A mixture of methanol and ultrapure water containing 1 formic acid 50:50, vv was selected as the mobile phase. In the separation carried out under isocratic conditions, the retention time of HRM was determined as 5.5 min, and the entire analysis was completed in under 8 min. The method demonstrated excellent linearity within the concentration range of 136 µgmL, with a correlation coefficient R² of 0.99998. The accuracy and precision parameters of the method were found to be within acceptable limits, with relative error RE and relative standard deviation RSD both below 2. The limits of detection and quantification LOD and LOQ were 0.269 µgmL and 0.816 µgmL, respectively. Also, it was determined that HRM standard solutions were stable for up to 7 days when stored at different temperatures 20C, 28C, 25C. In conclusion, this method offers a reliable alternative for the quantitative determination of HRM with high analytical quality, reproducibility, and short time. It can be widely used in routine laboratory practices for the monitoring of HRM in pure form and in various pharmaceutical formulations.

Article Details

Development and Validation of a Simple HPLC-UV Method for Analysis of Harmine

Authors:

Galip Mesut Demir | Afife Büşra Uğur Kaplan | Mine Gülaboğlu | Meltem Çetin

Publisher:

Karyay Karadeniz Yayımcılık

Main Topics:

Tıbbi ve Biyomoleküler Kimya (Diğer)

Journal:

Black Sea Journal of Engineering and Science Vol.8 Issue 4, pp.1096-1102

Document Type:

Article

Publication Date:

7/2025

Document Language:

English

Document ID (Org No.):

2001346265

pISSN:

2619-8991

Harmine

HPLC-UV method

Peganum harmala

Stability

ValidationHarmine

HPLC-UV method

Peganum harmala

Stability

Validation

Issues

Article Details

Authors: Galip Mesut Demir | Afife Büşra Uğur Kaplan | Mine Gülaboğlu | Meltem Çetin

Publisher: Karyay Karadeniz Yayımcılık

Main Topics: Tıbbi ve Biyomoleküler Kimya (Diğer)

Journal: Black Sea Journal of Engineering and Science Vol.8 Issue 4, pp.1096-1102

Document Type: Article

Publication Date: 7/2025

Document Language: English

Document ID (Org No.): 2001346265

pISSN: 2619-8991

Issues

Authors:

Galip Mesut Demir | Afife Büşra Uğur Kaplan | Mine Gülaboğlu | Meltem Çetin

Publisher:

Karyay Karadeniz Yayımcılık

Main Topics:

Tıbbi ve Biyomoleküler Kimya (Diğer)

Journal:

Black Sea Journal of Engineering and Science Vol.8 Issue 4, pp.1096-1102

Document Type:

Article

Publication Date:

7/2025

Document Language:

English

Document ID (Org No.):

2001346265

pISSN:

2619-8991