THẨM ĐỊNH PHƯƠNG PHÁP ĐỊNH LƯỢNG ĐỒNG THỜI CAFFEIN VÀ ACID CHLOROGENIC TRONG HẠT CÀ PHÊ XANH BẰNG PHƯƠNG PHÁP SẮC KÝ LỎNG HIỆU NĂNG CAO

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Tác giả: Hoàng Anh Vĩnh Hà, Thị Thu Huyền Hoàng, Văn Chung Huỳnh, Trung Khoảng Lê, Lưu Phúc Trần

Ngôn ngữ: vie

Ký hiệu phân loại:

Thông tin xuất bản: Tạp chí Khoa học và Công nghệ - Đại học Thái Nguyên, 2023

Mô tả vật lý: tr.308-315

Bộ sưu tập: Metadata

ID: 339788

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Nghiên cứu được tiến hành nhằm mục đích thẩm định qui trình định lượng đồng thời caffein và acid chlorogenic trong hạt cà phê bằng phương pháp sắc ký lỏng hiệu năng cao. Quá trình phân tích được thực hiện trên máy HPLC Waters e2695, cột sắc ký Reliant C18 (5µm
4,6x250 mm), pha động là acid formic 0,1% trong nước (A) và acetonitril (B). Dung môi pha động được chạy theo gradient nồng độ: Thời điểm ban đầu 5% B, tăng dần tới phút 20 là 10% B rồi giữ cố định cho tới phút 30 phút. Thể tích bơm mẫu là 10 µl. Tốc độ dòng 1 ml/phút. Đo ở 2 bước sóng 270 nm và 330 nm. Quy trình định lượng đã được thẩm định đáp ứng các yêu cầu của ICH (International Conference on Harmonization). Kết quả thu được cho thấy, phương pháp đạt yêu cầu với các chỉ tiêu: độ đặc hiệu, khoảng tuyến tính (R2 >
0,99), độ lặp lại cao với RSD <
0,7% đối với diện tích peak và RSD <
0,45 % đối với thời gian lưu, giới hạn phát hiện (≥ 0,06 µg/ml cho CAF và 0,36 µg/ml cho CGA), giới hạn định lượng (≥ 0,19 µg/ml cho CAF và 1,10 µg/ml cho CGA). Độ đúng của phương pháp cũng đạt yêu cầu thông qua chỉ số độ thu hồi nằm trong khoảng 97,31 – 104,26%. Do vậy, phương pháp định lượng phù hợp để xác định đồng thời hàm lượng caffein và acid chlorogenic trong hạt cà phê.The aim of study is to validate a quantitative method of caffeine (CAF) and chlorogenic acid (CGA) contents in green coffee bean by high performance liquid chromatography (HPLC). The analysis was performed on HPLC Waters e2695, Reliant C18 column (5 µm
4.6x250 mm), using the mobile phase of formic acid 0.1% in water (A) and acetonitrile (B). The mobile phase gradients were operated at: initially 5% B, gradually increased to 10% B by 20 mins and remained constant until 30 mins. The sample injection volume was 10 µl. Flow rate was 1 ml/min. Measurement was taken at two wave lengths at 270 nm and 330 nm. The quantification process has been validated to meet the requirements of the ICH (International Conference on Harmonization). Results show that the method meets criteria such as specificity, linearity (R2 >
0.99), high repeatability with RSD <
0.7% for peak area and RSD <
0.45% for retention time, limit of detection (≥ 0.06 µg/ml for CAF and 0.36 µg/ml for CGA), limit of quantification (≥ 0.19 µg/ml for CAF and 1.10 µg/ml for CGA). The accuracy of the method also meets requirements through the recovery rate ranging from 97.31 – 104.26%. Thus, the quantification method is suitable for simultaneous determination of caffeine and chlorogenic acid in green coffee beans.

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1. Đọc trực tuyến

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