Identification of efficacy components and determination of isomaltooligosaccarides in naobaijin oral solution

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1 8 5 Vol. 8 No Journal of Food Safety and Quality May., 2017 张连龙, 杨月 *, 周华生, 毛兆祥, 郭去非, 成恒嵩, 任会娜 (, ) 摘要 : 目的 方法 (solid-phase extraction, SPE) 结果 ; ; 结论, 关键词 : ; ; ; ; Identification of efficacy components and determination of isomaltooligosaccarides in naobaijin oral solution ZHANG Lian-Long, YANG Yue *, ZHOU Hua-Sheng, MAO Zhao-Xiang, GUO Qu-Fei, CHENG Heng-Song, REN Hui-Na (Wuxi Giant Pharmaceutical Co., Ltd., Wuxi , China) ABSTRACT: Objective To investigate the methods for identification of efficacy components and determination of isomaltooligosaccarides in naobaijin oral solution. Methods Precipitation, centrifugal, solid-phase extraction (SPE) column separation, enrichment and purification methods were used for sample pretreatment. The contents of isomaltooligosaccharides, hawthorn and poria in naobaijin oral solution were identified and determined by chemical reaction, thin-layer chromatography (TLC) and high performance liquid chromatography (HPLC). The content of isomaltooligosaccarides was determined by HPLC of double column. Results Ethanol precipitation method, anthrone method and phenol sulfuric acid methods effectively identified the isomaltooligosaccharides, potassium permanganate fade method, TLC and HPLC methods effectively identified citric acid in hawthorn, and Folin-phenol reagent method and ninhydrin color method effectively identified water soluble protein and amino acids in poria. Conclusion The above methods have strong specificity, good reproducibility, and high sensitivity, which can be used to ensure the quality of the products. KEY WORDS: naobaijin oral solution; health food; efficacy components; identification; content determination * 通讯作者 :,, dtyue@126.com *Corresponding author: YANG Yue, Engineer, Wuxi Giant Pharmaceutical Co., Ltd., Wuxi , China. dtyue@126.com

2 5, : 引言,, (2016 ), :,, [1] 2015 [2],, [1,2],,,, 2 材料与方法 2.1 试验材料 ( , ); ( HBL ); ( ) G(10 cm 20 cm, ); (solid-phase extraction, SPE) C18-SCX C8-SCX HLB-SCX MAX( Tegent ) (, ); ; 2.2 试验仪器 2695 ( Waters ); AE-240 ( Mettler ); TGL-10B ( ) 2.3 试验方法 低聚异麦芽糖的鉴别 [3] 山楂的鉴别 (1) [4] : 20 ml, 80 ml,,, 6000 r/min, 30 min, ( ) : 0.25 g, 50 ml, 30 min,, : 5 g/l :, 4 2 ml, 10 ml, 3 [5], 1~2 min,, 0.02 mol/l, (2) [5,6,7] : C18-SCX HLB-SCX : 6 ml, 0.1 mol/l 6 ml ; : 60 ml,, 5 ml, 0.1 mol/l 5 ml ; : 0.1 mol/l 6 ml, ; : 10 ml - (9:1, V:V),,, : 0.5 g, 50 ml, 30 min,,, 5 ml, :,, 2 mg, : 1 ml, G, : (5:2:1.5:0.5, V:V:V:V); : 0.1% - (,, 10 min,, ) (3) (high performance liquid chromatography, HPLC) ) [6] : 40 ml,, 5 ml : 1 g, 50 ml, 30 min, : 0.4 g/l μm, Lichraspher C 18 (4.6 mm 250 mm, 5 μm); : mol/l (5:95, V:V), ph 2.5; : 40 ; : 1.0 ml/min; : 210 nm; : 20 μl 茯苓的鉴别 (1) [5] : 1 ml, 10 ml : 0.5 g, 25 ml, min,, 3~5 ml,

3 ml, 10 ml :, 3 A [5] 5 ml, 10 min, B [5] 0.5 ml, 30 min (2) [3] : C8-SCX : 6 ml, 10 ml ; : 50 ml,, 5 ml, 0.1 mol/l 5 ml ; : 0.1 mol/l 5 ml 5 ml, ; : 5% - 10 ml,, : 0.5 g, 20 ml, 30 min,,, 5 ml, :, [3] : 3 1 ml, 3 3~5,,,, 2~3 [5],, 80 5~10 min [3] : 3 1 ml, 10 ml, ph 8.0 ( Na 2 HPO 4 12H 2 O g KH 2 PO g, 100mL)0.5 ml,, [5] 0.5 ml, ~15 min 低聚异麦芽糖含量的测定 [1] [8] MAX 1 ml 10 ml,, 1 ml,,, 0.22 μm,, -, (Sugarpak 1),, (refractive index detector, RID), 85, 0.4 ml/min, 10 μl 3 结果与分析 3.1 鉴别低聚异麦芽糖,,,,, [6] 5%,, 3.2 鉴别山楂 高锰酸钾褪色法, :,, 3, Mn 7+ ( ) Mn 2+ ( ) [9] 薄层色谱法 7, 1,,,,,,, HLB-SCX C18-SCX 2, 2 [10,11], HLB-SCX C18-SCX 3, Fig. 1 Thin layer chromatogram of citric acid 1. C18-SCX; 2. HLB-SCX; 3. C18-SCX; 4. HLB-SCX; 5. ; 6. C18-SCX; 7. HLB-SCX 高效液相色谱法, 2 2,,, 5 min,,,,,,,,,, SPE,

4 5, : Fig. 2 HPLC chromatograms of citric acid

5 脑白金口服液中茯苓的鉴别 福林酚试剂法鉴别茯苓中的蛋白质 3 A,,,,,, 茚三酮显色法鉴别茯苓中的氨基酸 [3] [3], 3~4 h,,, C8-SCX 3.4 脑白金口服液中低聚异麦芽糖含量的测定 3 4, ( 3),, ; Sugarpak 1,,,, ( 4) [1] [8], 3 Hypersil APS2 Fig.3 Chromatogram of isomaltooligosaccharide in the Hypersil APS2 column 4 Sugarpak 1 Fig. 4 Chromatogram of isomaltooligosaccharide in the Sugarpak 1 column

6 5, : 1677 表 1 3 批脑白金口服液样品的检测结果 Table 1 Results of 3 batches of naobaijin oral solution (g/100 ml) (g/100 ml) (g/100 ml) (g/100 ml) (g/100 ml) (g/100 ml) 低聚异麦芽糖含量的测定,, 3, 1 4 讨论与结论,, 40% [2],,,,,, SPE [12,13] [14],, [14] ; [8],, C18-SCX HLB-SCX ; C8-SCX ; MAX, [15] 3 SPE,,, [16], SPE,,,,,,,,,,, SPE,, SPE,,,, 参考文献 [1] GB/T [S]. GB/T Isomaltooligsaccharide [S]. [2]. ( )[M]. :, Chinese Pharmacopoeia Commission. Chinese pharmacopoeia (part one) [M]. Beijing: Chinese Medical Science And Technology Press, [3],,. [M]. :, Mi XL, Niu WM, Cheng HS. Quality analysis technology of health food [M]. Nanjing: Jiangsu Science and Technology Press, [4]. ( )[M]. :, Chinese Pharmacopoeia Commission. Chinese pharmacopoeia (part two) [M]. Beijing: Chinese Medical Science And Technology Press, [5]. ( )[M]. :, Chinese Pharmacopoeia Commission. Chinese pharmacopoeia (part four) [M]. Beijing: Chinese Medical Science And Technology Press, [6],,,. [J]., 2008, 19(8): He B, Feng WY, Tian J, et al. TLC Identification and content determination of citric acid in huangjin yanhou tablet [J]. Lishizhen Med Mater Med Res, 2008, 19(8): [7] Renzo B, Marina M, Monica A, et al. Rapid mixed mode solid phase extraction method for the determination of acrylamide in roasted coffee by HPLC-MS/MS [J]. Food Chem, 2012, 135(4): [8],,,. [C]., Chen SW, Zhu S, Zhang LL, et al. Determination of isomaltooligsaccharides in naobaijin oral liquid [C]. The sixth academic papers of HPLC and MS in east china area, [9]. [M]. :, Tianjin University edited. Organic chemistry [M]. Beijing: People's

7 Education Press, [10],. [M]. :, Chen XH, Wang QJ. Solid phase extraction technology and application [M]. Beijing: Science Press, [11] Nostrum CE. Solid phase extraction: A new tool for drug analysis [J]. Drug Technol Today, 2005, 2(1): [12] Boguslaw B, Malgorzata S. Past, present, and future of solid phase extraction: A review [J]. Crit Rev Anal Chem, 2012, 42(3): [13] Spietelun A, Marcinkowski T, Dela GM, et al. Recent developments and future trends in solid phase microextraction techniques towards green analytical chemistry [J]. J Chromatogr, 2013, 1321: [14] De Martinis BS, Barnes A, Scheidweiler KB, et al. Development and validation of a disk solid phase extraction and gas chromatography-mass spectrometry method for MDMA [J]. J Chromatogr B, 2007, 852(1): [15]. - [J]., 2007, 26(2): Fu RN. Advances on SPE-chromatography in China in recent years [J]. Chin J Anal Lab, 2007, 26(2): [16] Azadmard-Damirchi S, Nemati M, Hesari J, et al. Rapid separating and enrichment of 4,4'-dimethylsterols of vegetable oils by solid-phase extraction [J]. J Am Oil Chem Soc, 2010, 87(10): 作者简介 ( 责任编辑 : 姜姗 ) 张连龙, 硕士, 研究员级高级工程师, 主要研究方向为药品 保健食品的研制与产品质量分析方法的研究 @qq.com 杨月, 硕士, 工程师, 主要研究方向为药品 保健食品的研制与产品质量分析方法的研究 dtyue@126.com