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Detection and Estimation of Vitamin D2 in Catharanthus Roseus by HPLC and other Molecular Spectra Instruments as A Source of Vitamin D3 Production

Authors: Mohammed Abbood Ayyash --- Mufeed Jalil Ewadh --- Nisreen Jalal Mohammed
Journal: Medical Journal of Babylon مجلة بابل الطبية ISSN: 1812156X 23126760 Year: 2016 Volume: 13 Issue: 2 Pages: 468 -480
Publisher: Babylon University جامعة بابل

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Abstract

Vitamin D is one of fat-soluble vitamins, considered a steroid hormone. It is regulate and responsible for improving the metabolism and absorption of calcium, iron, magnesium, zinc, and phosphate in small intestine.SHIMADZU High-Performance Liquid Chromatography was used for qualitative and quantitative estimation of vitamin D2 in aqueous extracted of Catharanthus Roseus plant; (20µl, 2.5 IU/gm) standard material of pure vitamin D2 (Ergocalciferol) injected and measured at reference conditions to fix retention time (RT), then 20µlof extracted plant sample was injected and measured at the same conditions. An intensive analysis conducted on aqueous, alcoholic extract and dry powder of plant leaves using modern and sophisticated instruments. Quantitative results from HPLC shows thatVit.D2 in drying weight of plant up to 187.840 IU/gm. Qualitative results from HPLC, UV-VIS, IR, ATR-FTIR shows match the effective groups of pure standard Vit.D2 and alcoholic extract, dry powder of C. Roseus plant.This study aimed to investigate the presence of vitamin D2 in C. Roseus, in addition to qualitative and quantitative detection in aqueous, alcoholic extract, and dry powder of plant leavesto use it in practical as a major source for Vit. D3 production in industrial scale; in order to use it in the medical, pharmaceutical and what is called alternative medicine (Herbs) fields.


Article
Development of A New HPLC Method for Separation and Determination 25(OH)- Cholecalciferol (Vitamin D3).
تطویر طریقة جدیدة في كروماتوكرافیا السائل عالي الأداء لفصل وتقدیر (D 25- فیتامین ( 3 (OH)-Cholecalciferol

Author: Sarmad B. Dikran سرمد بهجت ديكران
Journal: journal of al-qadisiyah for pure science(quarterly) مجلة القادسية للعلوم الصرفة (فصلية). ISSN: 19972490 Year: 2016 Volume: 4 Issue: 21 Pages: 101-111
Publisher: Al-Qadisiyah University جامعة القادسية

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Abstract

Vitamin D3 was found interference with other fat soluble vitamins (especially vitamin A and E) that arecommonly present with vitamin D, in Serum & pharmaceutical preparation.In the present study, a rapid, simple and economical reversed phase-HPLC procedure has been fixedoptimization of condition and developed for the separation and determination of vitamin D3 in mixture standardof Vitamin D3and Vitamin A.Anew method were developed in HPLC ,based method with a UV detector by examining various conditionsincluding mobile phase, flow rate, volume inject and temperature, we have also proposed a very simple methodof isolation of these vitamins. The vitamins were separated isocratically on a Knauer C18 column (250×4.6mm,5μm particle size) with a mobile phase consisting of isocratic acetonitrile and methanol (75:25, v/v) operated at40ºC with retention times less than 10 min. and the detection limit of our developed HPLC method is 0.01μg/ml.The eluted vitamins were identified and monitored on a UV-Detector at 265 nm with optimum flow rate 1.5mL/min and 50μL Injection volume. The linearity of the method was excellent (r2 > 0.999), over the concentration rangeof 10-200ng/ml.Vitamin D3 was found interference with other fat soluble vitamins (especially vitamin A and E) that arecommonly present with vitamin D, in Serum & pharmaceutical preparation.In the present study, a rapid, simple and economical reversed phase-HPLC procedure has been fixedoptimization of condition and developed for the separation and determination of vitamin D3 in mixture standardof Vitamin D3and Vitamin A.Anew method were developed in HPLC ,based method with a UV detector by examining various conditionsincluding mobile phase, flow rate, volume inject and temperature, we have also proposed a very simple methodof isolation of these vitamins. The vitamins were separated isocratically on a Knauer C18 column (250×4.6mm,5μm particle size) with a mobile phase consisting of isocratic acetonitrile and methanol (75:25, v/v) operated at40ºC with retention times less than 10 min. and the detection limit of our developed HPLC method is 0.01μg/ml.The eluted vitamins were identified and monitored on a UV-Detector at 265 nm with optimum flow rate 1.5mL/min and 50μL Injection volume. The linearity of the method was excellent (r2 > 0.999), over the concentration rangeof 10-200ng/ml.Key Words: 25(OH) Cholecalciferol (Vitamin D3) , HPLC..

یتواجد فیتامین 3والمستحضرات الدوائیة . أذ تم في ھذه الدراسة الحالیة استخدام طریقة سریعة وبسیطة واقتصادیة وحساسة ھي كروموتوغرافیا السائل عاليفي مزیج یحوي محالیل قیاسیة لكل من D وتثبیت ظروف قیاسیة مثلى جدیدة لفصل وتقدیر فیتامین 3 reversal HPLC الاداء ذو الطور العكوس. D و 3 A فیتامین200-400 ) ، الطور المتحرك، سرعة ) nm وشملت الدراسة الظروف المثلى التي ثبت بھا الفصل ضمن مدى من الاطول الموجیة تراوحت مابینالجریان، حجم النموذج المحقون ودرجة الحرارة أذ تم فصل ھذه الفیتامینات بطریقة سھلة وبأستخدام عمود فصل ذو مواصفاتوطور متحرك مكون من مزیج الایثانول: Knauer/Germany والمجھز من شركة C18 column (250×4.6mm, 5μm particle size)0.01 . وتم تشخیص μg/mL 40 وبزمن تظھیر اقل من 10 دقائق وحد كشف عند ºC الاسیتونایترایل بنسبة حجمیة ( 25:75 )ودرجة حرارة265 و بمعدل سرعة جریان مثلى 1.5 nm عند طول موجي UV ومراقبة الفیتامینات المعزولة بأستخدام مكشاف الاشعة فوق البنفسجیة.50 μl و كان حجم النموذج المحقون mL/min

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