zldmЗаводская лаборатория. Диагностика материаловIndustrial laboratory. Diagnostics of materials1028-68612588-0187ООО «Издательство «ТЕСТ-ЗЛ»10.26896/1028-6861-2019-85-1-I-18-27zldm-868Research ArticleАНАЛИЗ ВЕЩЕСТВАSUBSTANCES ANALYSISСтандартизация экстрактов корней P. ginseng и P. quinquefolius методом жидкостной хромато-масс-спектрометрииStandartization of P. ginseng and P. quinquefolius root extracts by liquid chromatography-mass spectrometryФедороваЕ. С.FedorovaE. S.

Москва

Department of Chemistry, Moscow

СтекольщиковаЕ. А.StekolshchikovaE. A.

Москва

Department of Chemistry, Moscow

ШпигунО. А.ShpigunO. A.

Москва

Department of Chemistry, Moscow

РодинИ. А.RodinI. A.

Москва

Department of Chemistry, Moscow

СтаврианидиА. Н.StavrianidiA. N.

Москва

Department of Chemistry, Moscow

Stavrianidi.andrey@gmail.comМосковский государственный университет имени М. В. ЛомоносоваРоссияLomonosov Moscow State UniversityRussian Federation201913022019851(I)1827Copyright © Федорова Е.С., Стекольщикова Е.А., Шпигун О.А., Родин И.А., Ставрианиди А.Н., 20192019Федорова Е.С., Стекольщикова Е.А., Шпигун О.А., Родин И.А., Ставрианиди А.Н.Fedorova E.S., Stekolshchikova E.A., Shpigun O.A., Rodin I.A., Stavrianidi A.N.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.zldm.ru/jour/article/view/868

С использованием метода высокоэффективной жидкостной хромато-масс-спектрометрии (ВЭЖХ-МС) разработан подход к определению тритерпеновых гликозидов в экстрактах женьшеня. Повышение селективности по сравнению с используемыми для стандартизации экстрактов из растительного сырья ВЭЖХ-УФ методиками позволило одновременно идентифицировать и определять 23 основных и минорных гинсенозида. Для этого помимо применения высокоселективного МС детектирования ионов аддуктов молекул гинсенозидов с натрием и фрагментных ионов сапогенина, были специально подобраны условия для хроматографического разделения на сорбенте с привитыми пентафторфенильными группами. Отдельно исследовали влияние температуры и состава подвижной фазы на селективность разделения определяемых гликозидов. Для некоторых пар соединений (F4 и Rg6, Rk3 и Rh4) добиться полного хроматографического разделения пиков не удалось, однако их определение при совместном присутствии возможно благодаря регистрации различающихся по отношению m/z сигналов. Для всех определяемых соединений были установлены диапазоны линейности и уравнения градуировочных зависимостей, а также определены метрологические характеристики, такие как предел обнаружения и воспроизводимость. Разработанный подход был опробован при стандартизации референтных экстрактов корней азиатского (P. ginseng) и американского (P. quinquefolius) женьшеня. Для некоторых гинсенозидов заявленные производителем содержания не соответствовали действительным, для остальных определенные концентрации оказались близки к заявленным. Кроме того, удалось расширить круг контролируемых гинсенозидов, что важно при медицинском применении таких экстрактов.

 

An approach to detection of triterpene glycosides in ginseng extracts is developed using high performance liquid chromatography/mass spectrometry (HPLC-MS). Enhanced selectivity compared to commonly used HPLC-UV techniques provides simultaneous registration of chromatographic peaks and determination of 23 major and minor ginsenosides. For this purpose, in addition to the use of highly selective MS detection of adduct ions of sodium ginsenoside molecules and fragmentary sapogenin ions, special conditions of chromatographic separation on a sorbent modified with pentafluorophenyl (PFP) groups were specified. The effect of column temperature and mobile phase composition on the separation selectivity of glycosides was also studied. Though we failed to achieve complete chromatographic separation of the peaks for several compounds (F4 and Rg6, Rk3 and Rh4), their determination appeared possible in case of their joint attendance due to registration of the signals that differ in the value of m/z ratio. For all studied compounds, the linearity ranges and calibration equations along with the metrological characteristics such as the detection limit and reproducibility were determined. The developed approach was tested in standardization of the reference extracts of Asian (P. ginseng) and American (P. quinquefolius) ginseng roots. For some ginsenosides, the content declared by the manufacture did not match the actual content, but for others the concentrations were close to the declared values. Moreover, we managed to expand the range of ginsenosides under control which is rather important for their use in medicine.

высокоэффективная жидкостная хроматографиямасс-спектрометрияконтроль качестварастительные экстрактыгинсенозидыселективностьпентафторфенильные заместителиhigh performance liquid chromatographymass spectrometryquality controlplant extractsginsenosidesselectivitypentafluorophenyl substituentsРФФИReferencesDharmananda S. The nature of ginseng: traditional use, modern research and the question of dosage / HerbalGram. 2002. Vol. 54. P. 34 – 51.Dharmananda S. The nature of ginseng: traditional use, modern research and the question of dosage / HerbalGram. 2002. Vol. 54. P. 34 – 51.ESCOP Monographs. European Scientific Cooperative on Phytotherapy. 2nd ed. — D.: Exeter, 2003. — 211 p.ESCOP Monographs. European Scientific Cooperative on Phytotherapy. 2nd ed. — D.: Exeter, 2003. — 211 p.Kiefer D., Pantuso T. Panax ginseng / Am. Family Phys. 2003. Vol. 68. N 8. P. 1539 – 1542.Kiefer D., Pantuso T. Panax ginseng / Am. Family Phys. 2003. Vol. 68. N 8. P. 1539 – 1542.Yang W. Z., Ye M., Qiao X., et al. A strategy for efficient discovery of new natural compounds by integrating orthogonal column chromatography and liquid chromatography/mass spectrometry analysis: Its application in Panax ginseng, Panax quinquefolium and Panax notoginseng to characterize 437 potential new ginsenosides / Anal. Chim. Acta. 2012. Vol. 739. P. 56 – 66.Yang W. Z., Ye M., Qiao X., et al. A strategy for efficient discovery of new natural compounds by integrating orthogonal column chromatography and liquid chromatography/mass spectrometry analysis: Its application in Panax ginseng, Panax quinquefolium and Panax notoginseng to characterize 437 potential new ginsenosides / Anal. Chim. Acta. 2012. Vol. 739. P. 56 – 66.Mathon C., Duret M., Kohler M., et al. Multi-targeted screening of botanicals in food supplements by liquid chromatography with tandem mass spectrometry / Food Chem. 2013. Vol. 138. N 1. P. 709 – 717.Mathon C., Duret M., Kohler M., et al. Multi-targeted screening of botanicals in food supplements by liquid chromatography with tandem mass spectrometry / Food Chem. 2013. Vol. 138. N 1. P. 709 – 717.Stavrianidi A. N., Rodin I. A., Braun A. V., et al. Simultaneous determination of ginsenosides by high-performance liquid chromatography with tandem mass spectrometry detection / J. Anal. Chem. 2014. Vol. 69. N 13. P. 1252 – 1258.Stavrianidi A. N., Rodin I. A., Braun A. V., et al. Simultaneous determination of ginsenosides by high-performance liquid chromatography with tandem mass spectrometry detection / J. Anal. Chem. 2014. Vol. 69. N 13. P. 1252 – 1258.Li W. K., Gu C. G., Zhang H. J., et al. Use of high performance liquid chromatography-tandem mass spectrometry to distinguish Panax ginseng C. A. Meyer (Asian ginseng) and Panax quinquefolius L. (North American ginseng) / Anal. Chem. 2000. Vol. 72. N 21. P. 5417 – 5422.Li W. K., Gu C. G., Zhang H. J., et al. Use of high performance liquid chromatography-tandem mass spectrometry to distinguish Panax ginseng C. A. Meyer (Asian ginseng) and Panax quinquefolius L. (North American ginseng) / Anal. Chem. 2000. Vol. 72. N 21. P. 5417 – 5422.Wan J. B., Li S. P., Chen J. M., Wang Y. T. Chemical characteristics of three medicinal plants of the Panax genus determined by HPLC-ELSD / J. Sep. Sci. 2007. Vol. 30. N 6. P. 825 – 832.Wan J. B., Li S. P., Chen J. M., Wang Y. T. Chemical characteristics of three medicinal plants of the Panax genus determined by HPLC-ELSD / J. Sep. Sci. 2007. Vol. 30. N 6. P. 825 – 832.Chan T. W., But P. P., Cheng S. W., et al. Differentiation and authentication of Panax ginseng, Panax quinquefolius and ginseng products by the use of HPLC/MS / Anal. Chem. 2000. Vol. 72. N 6. P. 1281 – 1287.Chan T. W., But P. P., Cheng S. W., et al. Differentiation and authentication of Panax ginseng, Panax quinquefolius and ginseng products by the use of HPLC/MS / Anal. Chem. 2000. Vol. 72. N 6. P. 1281 – 1287.The United States Pharmacopeia 30th edn. — Rockville, MD: United States Pharmacopeial Convention, Inc., 2007.The United States Pharmacopeia 30th edn. — Rockville, MD: United States Pharmacopeial Convention, Inc., 2007.Lau A. J., Seo B. H., Woo S. O., Koh H. L. High-performance liquid chromatographic method with quantitative comparisons of whole chromatograms of raw and steamed Panax notoginseng / J. Chromatogr. A. 2004. Vol. 1057. N 1 – 2. P. 141 – 149.Lau A. J., Seo B. H., Woo S. O., Koh H. L. High-performance liquid chromatographic method with quantitative comparisons of whole chromatograms of raw and steamed Panax notoginseng / J. Chromatogr. A. 2004. Vol. 1057. N 1 – 2. P. 141 – 149.Li L., Zhang J. L., Sheng Y. X., et al. Simultaneous quantification of six major active saponins of Panax notoginseng by high-performance liquid chromatography-UV method / J. Pharm. Biomed. Anal. 2005. Vol. 38. N 1. P. 45 – 51.Li L., Zhang J. L., Sheng Y. X., et al. Simultaneous quantification of six major active saponins of Panax notoginseng by high-performance liquid chromatography-UV method / J. Pharm. Biomed. Anal. 2005. Vol. 38. N 1. P. 45 – 51.Kim S. N., Ha Y. W., Shin H., et al. Simultaneous quantification of 14 ginsenosides in Panax ginseng C. A. Meyer (Korean red ginseng) by HPLC-ELSD and its application to quality control / J. Pharm. Biomed. Anal. 2007. Vol. 45. N 1. P. 164 – 170.Kim S. N., Ha Y. W., Shin H., et al. Simultaneous quantification of 14 ginsenosides in Panax ginseng C. A. Meyer (Korean red ginseng) by HPLC-ELSD and its application to quality control / J. Pharm. Biomed. Anal. 2007. Vol. 45. N 1. P. 164 – 170.Sun B. S., Gu L. J., Fang Z. M., et al. Simultaneous quantification of 19 ginsenosides in black ginseng developed from Panax ginseng by HPLC – ELSD / J. Pharm. Biomed. Anal. 2009. Vol. 50. N 1. P. 15 – 22.Sun B. S., Gu L. J., Fang Z. M., et al. Simultaneous quantification of 19 ginsenosides in black ginseng developed from Panax ginseng by HPLC – ELSD / J. Pharm. Biomed. Anal. 2009. Vol. 50. N 1. P. 15 – 22.Cui M., Song F., Zhou Y., et al. Rapid identification of saponins in plant extracts by electrospray ionization multi-stage tandem mass spectrometry and liquid chromatography/tandem mass spectrometry / Rapid. Commun. Mass Spectrom. 2000. Vol. 14. N 14. P. 1280 – 1286.Cui M., Song F., Zhou Y., et al. Rapid identification of saponins in plant extracts by electrospray ionization multi-stage tandem mass spectrometry and liquid chromatography/tandem mass spectrometry / Rapid. Commun. Mass Spectrom. 2000. Vol. 14. N 14. P. 1280 – 1286.

The authors declare that there are no conflicts of interest present.